benzofurans and 3-n-butylphthalide

Inflammation and immunity play an essential role in disease pathogenesis. 3-N-Butylphthalide (NBP), a group of compounds extracted from seeds of

Topics: Anti-Inflammatory Agents; Benzofurans; Humans; Inflammation; Neuroprotective Agents

Current evidence for the efficacy of pharmacological treatment in improving cognitive function is absent. Recent studies have reported that 3-n-butylphthalide (NBP) has a positive effect on improving cognitive impairment; however, its clinical efficacy and safety is unclear. Therefore, we conducted a meta-analysis to assess its efficacy and safety for cognitive impairment.. We systematically searched the PubMed, EMBASE, Cochrane Library, Web of Science, and Scopus databases, and two reviewers independently screened and extracted the data from included studies. We synthesized the data using the Review Manager Software version 5.3.. We included six randomized clinical trials (RCTs), encompassing 851 patients with cognitive impairment. The results showed that NBP improved cognitive impairment. Specifically, the clinical efficacy was better than that in the control group, with better performance in improving the Mini-Mental State Examination and the Montreal Cognitive Assessment scores, while decreasing the Alzheimer's Disease Assessment Scale-Cognitive subscale and the Clinician's Interview-Based Impression of Change plus caregiver input scores. There was no significant difference in the incidence of adverse events between both groups.. The NBP is effective and safe in improving cognitive impairment; however, more high-quality RCTs are needed to confirm these findings.

Topics: Benzofurans; Cognition; Cognition Disorders; Cognitive Dysfunction; Humans

3-N-butylphthalide (NBP) is a component isolated from seeds of Chinese celery, and it was firstly approved for the treatment of ischemic stroke. With the gradual in-depth understanding of its pharmacological action, it was found that it may have potential effects on treating diabetes and its complications. This review aims to illustrate the researches on the properties of NBP and its therapeutic efficacy in diabetic related diseases. This review will discuss the results of experiments in vitro and in vivo to make progress in understanding the beneficial effects of NBP and its derivatives on diabetic complications including diabetic vascular diseases, diabetic peripheral neuropathy, diabetic brain related diseases and diabetic cataract. We will also demonstrate NBP's numerous molecular targets and interactions with multiple cellular signaling pathways such as oxidative stress, inflammatory responses, apoptosis and autophagy. NBP is proved to be a potential therapeutic approach for treating diabetic complications.

Topics: Benzofurans; Diabetes Complications; Diabetes Mellitus; Humans; Oxidative Stress

Cerebral ischemia is also known as ischemic stroke. In recent years, research on neuroprotection after ischemia has became a hot spot as stroke can result in symptoms of nerve damages such as hemiplegia, learning and memory disorders. The key factors that cause the death of cells include excitotoxicity, oxidative damage, nitrosative stress and inflammation. However, there is no effective preparation for the treatment of post-ischemic nerve defects at present, so it is urgent to find and develop effective drugs for the treatment of nerve damages after ischemia. Traditional Chinese medicine has advantages and potentials in the treatment of neurological diseases. Many scholars have carried out related researches on the active ingredients of traditional Chinese medicine and achieved some good results. In this context, the researches on the neuroprotective effects of traditional Chinese medicines such as tetramethylpyrazine, butylphthalide and total saponins of Panax notoginseng were reviewed. The author found that the neuroprotective researches of traditional Chinese medicine mostly focused on anti-apoptosis, anti-inflammatory and anti-oxidative stress, but those effects were not sounique to the nervous system. Furthermore, most ingredients of traditional Chinese medicine showed a poor water-soluble property. In view of the research status and existing problems of traditional Chinese medicine in nerve injury, the suggestions for the research and development of the potent neuroprotective agents were proposed in this study from the perspective of pharmacological mechanism research and preparation theory.

Topics: Benzofurans; Brain Ischemia; Cerebral Infarction; Drugs, Chinese Herbal; Humans; Medicine, Chinese Traditional; Neuroprotective Agents; Panax notoginseng; Pyrazines; Saponins

Approximately 10% to 30% patients develop delayed encephalopathy after acute CO poisoning (DEACMP). No specific treatment is available and poor prognosis is a characteristic of this disease. We aimed to evaluate the efficacy and safety of all therapies that have been tried in randomized controlled trial (RCT) for DEACMP.. We conducted a systematic search of the Cochrane, Embase, PubMed, and Web of Science databases.. Overall, 4 RCTs were identified in our study. Both hyperbaric oxygen (HBO) and mesenchymal stem cell (MSC) transplantation were effective in DEACMP, and MSC seemed to be superior to HBO. The addition of dexamethasone, N-butylphthalide, or XingZhi-YiNao granules into HBO, or butylphthalide into MSC could achieve better neurological recovery in DEACMP patients but did not significantly increase the incidence of adverse events.. Several therapies have shown positive results in treating DEACMP and need to be proven by further studies.

Topics: Benzofurans; Brain Diseases; Carbon Monoxide Poisoning; Dexamethasone; Drugs, Chinese Herbal; Humans; Hyperbaric Oxygenation; Mesenchymal Stem Cell Transplantation; Randomized Controlled Trials as Topic; Time Factors

Concussive head injury (CHI) is quite prevalent in military personnel leading to lifetime disability in more than 85% of cases. Other reasons of CHI include motor vehicle accident, fall or blunt trauma under various conditions. In United States of America (USA) alone more than 150k cases of head injury are added every year for which no suitable therapeutic strategies are still available. Thus, there is a need to expand our knowledge in treating CHI cases with novel therapeutic measures to enhance the quality of life of head injury victims. With recent advancements in nanodelivery of drugs for superior neuroprotective effects in neurological diseases, our laboratory is engaged in understanding the role of nanowired delivery of suitable drugs in treating CHI and other neurodegenerative diseases. DL-3-n-butylphthalide (NBP) is an extract of Chinese celery and is able to induce profound neuroprotection following ischemic stroke and other related neurological dysfunction. Thus, it is quite likely that synthetic NBP could have pronounced neuroprotective effects in CHI as well. We believe that nanodelivery of NBP have superior neuroprotection in CHI. In this review neuroprotective effects of nanowired delivery of NBP in CHI induced brain pathology is described. Our experimental observations show that nanowired delivery of NBP results in superior neuroprotection than the regular NBP in CHI. The probable mechanisms and functional significance of our finding in relation to military medicine is discussed based on our own investigations.

Topics: Animals; Benzofurans; Brain Concussion; Brain Edema; Humans; Nanomedicine; Nanowires; Neuroprotective Agents

DL-3-n-butylphthalide (NBP) is widely used as a neuroprotective drug for ischemic stroke in China. There is, however, no established evidence on its efficacy and safety for patients with ischemic stroke. We, therefore, conducted a systematic review and meta-analysis. Major databases were searched to identify randomized controlled trials that assessed the efficacy and safety of NBP on ischemic stroke, reporting outcomes among patients treated with NBP alone or combined with standard anti-ischemic stroke drugs vs. standard anti-ischemic stroke drugs. Continuous data were validated, extracted and synthesized of standardized mean differences (SMDs) by random effects models, while dichotomous data were validated, extracted and synthesized of relative risk (RR) by random effects models. Twelve randomized controlled trials involving 1160 patients were identified. Results suggested that NBP monotherapy is not superior to standard anti-ischemic stroke drugs based on the Barthel Index (SMD, 0.25; 95% CI

Topics: Aged; Benzofurans; Brain Ischemia; Databases, Bibliographic; Drug Therapy, Combination; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Phytotherapy; Randomized Controlled Trials as Topic; Stroke; Treatment Outcome

In recent years, L-3-n-butylphthalide (L-NBP) has been used for Parkinson disease dementia (PDD) to attenuate cognitive impairments in China. Therefore, we selected published and qualified clinical trials to conduct a systematic review and meta-analysis with the aim of assessing the effectiveness and safety of L-NBP in the treatment of PDD.. This systematic review and meta-analysis aimed to assess the effectiveness and safety of L-NBP in the treatment of PDD.. We searched PubMed, EMBASE, China National Knowledge Infrastructure, Chinese Scientific Journal Database (VIP database), and Wan-Fang Database to collect eligible articles. We calculated pooled estimates of odds ratios or the standard mean deviation with 95% confidence intervals.. Eight randomized controlled trials were included in our meta-analysis. Our meta-analysis showed that L-NBP combined with Western medicine (WM) had a better effect on improving cognitive dysfunction, the total effective rate, symptoms of Parkinson disease (PD), and activities of daily living function than WM alone. Regarding safety, no serious adverse events were observed in the experimental group.. We found that L-NBP as a complementary therapy may have a positive therapeutic effect for improving cognitive dysfunction, the total effective rate, symptoms of PD, quality of life, and the related serum factors in the treatment of PDD. Furthermore, L-NBP was a safe treatment for PDD. However, the findings of our meta-analysis may be influenced by the low quality of the included studies. We highlight the need to conduct trials with higher methodological quality.

Topics: Antiparkinson Agents; Benzofurans; Capsules; Dementia; Humans; Nootropic Agents; Parkinson Disease; Randomized Controlled Trials as Topic

The 3-N-butylphthalide (NBP) comprises one of the chemical constituents of celery oil. It has a series of pharmacologic mechanisms including reconstructing microcirculation, protecting mitochondrial function, inhibiting oxidative stress, inhibiting neuronal apoptosis, etc. Based on the complex multi-targets of pharmacologic mechanisms of NBP, the clinical application of NBP is increasing and more clinical researches and animal experiments are also focused on NBP. The aim of this review was to comprehensively and systematically summarize the application of NBP on neurologic diseases and briefly summarize its application to non-neurologic diseases. Moreover, recent progress in experimental models of NBP on animals was summarized.. Literature was collected from PubMed and Wangfang database until November 2018, using the search terms including "3-N-butylphthalide," "microcirculation," "mitochondria," "ischemic stroke," "Alzheimer disease," "vascular dementia," "Parkinson disease," "brain edema," "CO poisoning," "traumatic central nervous system injury," "autoimmune disease," "amyotrophic lateral sclerosis," "seizures," "diabetes," "diabetic cataract," and "atherosclerosis.". Literature was mainly derived from English articles or articles that could be obtained with English abstracts and partly derived from Chinese articles. Article type was not limited. References were also identified from the bibliographies of identified articles and the authors' files.. NBP has become an important adjunct for ischemic stroke. In vascular dementia, the clinical application of NBP to treat severe cognitive dysfunction syndrome caused by the hypoperfusion of brain tissue during cerebrovascular disease is also increasing. Evidence also suggests that NBP has a therapeutic effect for neurodegenerative diseases. Many animal experiments have found that it can also improve symptoms in other neurologic diseases such as epilepsy, cerebral edema, and decreased cognitive function caused by severe acute carbon monoxide poisoning. Moreover, NBP has therapeutic effects for diabetes, diabetes-induced cataracts, and non-neurologic diseases such as atherosclerosis. Mechanistically, NBP mainly improves microcirculation and protects mitochondria. Its broad pharmacologic effects also include inhibiting oxidative stress, nerve cell apoptosis, inflammatory responses, and anti-platelet and anti-thrombotic effects.. The varied pharmacologic mechanisms of NBP involve many complex molecular mechanisms; however, there many unknown pharmacologic effects await further study.

Topics: Animals; Benzofurans; Humans; Nervous System Diseases; Neuroprotective Agents; Oxidative Stress

Stroke is a leading cause of morbidity and mortality in both developed and developing countries all over the world. The only drug for ischemic stroke approved by FDA is recombinant tissue plasminogen activator (rtPA). However, only 2-5% stroke patients receive rtPAs treatment due to its strict therapeutic time window. As ischemic stroke is a complex disease involving multiple mechanisms, medications with multi-targets may be more powerful compared with single-target drugs. Dl-3-n-Butylphthalide (NBP) is a synthetic compound based on l-3-n- Butylphthalide that is isolated from seeds of Apium graveolens. The racemic 3-n-butylphthalide (dl- NBP) was approved by Food and Drug Administration of China for the treatment of ischemic stroke in 2002. A number of clinical studies indicated that NBP not only improved the symptoms of ischemic stroke, but also contributed to the long-term recovery. The potential mechanisms of NBP for ischemic stroke treatment may target different pathophysiological processes, including anti-oxidant, antiinflammation, anti-apoptosis, anti-thrombosis, and protection of mitochondria et al. Conclusion: In this review, we have summarized the research progress of NBP for the treatment of ischemic stroke during the past two decades.

Topics: Animals; Benzofurans; Brain Ischemia; Humans; Platelet Aggregation Inhibitors; Stroke

Discovering effective agents to slow or stop neurodegeneration is a challenging task. Over decades, only a few drugs were approved by Food and Drug Administration (FDA) and most ended in failure. The lessons learned have switched the strategy of drug discovery from designing highly selective ligands to a network pharmacology approach. This enables many natural products like butylphthalide (NBP) once again to be regarded as a valuable source of leads for drug discovery. In this review, we first start with the neuroprotective effects of NBPs on acute ischemic stroke, and later spread to their applications in major neurodegenerative diseases. The underlying mechanisms are also discussed in order to provide a direction for further study. Hopefully, this review could bring some new insights for drug development in this struggling field.

Topics: Animals; Benzofurans; Humans; Neurodegenerative Diseases; Neuroprotective Agents; Stroke

Cataract is one of the most important causes of blindness worldwide, with age-related cataract being the most common one. Agents preventing cataract formation are urgently required. Substantial evidences point out aggravated oxidative stress as a vital factor for cataract formation. Nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/Kelch-like erythroid-cell-derived protein with CNC homology (ECH)-associated protein 1 (Keap1) system is considered as one of the main cellular defense mechanisms against oxidative stresses. This review discusses the role of Nrf2 pathway in the prevention of cataracts and highlights that Nrf2 suppressors may augment oxidative stress of the lens, and Nrf2 inducers may decrease the oxidative stress and prevent the cataract formation. Thus, Nrf2 may serve as a promising therapeutic target for cataract treatment.

Topics: Aging; Antioxidants; Benzofurans; Blindness; Cataract; Flavonoids; Gene Expression Regulation, Developmental; Humans; Kelch-Like ECH-Associated Protein 1; Lens, Crystalline; NF-E2-Related Factor 2; Oxidative Stress; Quercetin; Signal Transduction; Transcription, Genetic

The research of alternative treatment for ischemic stroke and degenerative diseases has always been a priority in neurology. 3-N-Butylphthalide (NBP), a family of compounds initially isolated from the seeds of

Topics: Apium; Apoptosis; Benzofurans; Humans; Inflammation; Nervous System Diseases; Neuroprotective Agents; Oxidative Stress; Plant Extracts

Natural products have been used as medicinal agents for many years. In addition, these compounds have also served as the starting points for semisynthetic analogs with improved properties. This review focuses on recent advances in the pharmacological studies on natural products mainly performed and published in China. Emphasis will be placed on those compounds that show the greatest promise clinically such as huperzine A (9-amino-13-ethylidene-11-methyl-4-azatricyclo[7.3.1.0(3.8)]trideca-3(8),6,11-trien-5-one), s-(-)-3-n-butylphthalide (s-(-)-3-butyl-1(3H)-isobenzofuranone), (-)-clausenamide (3-hydroxy-4-phenyl-5a-hydroxybenzyl-N-methyl-gamma-lactam) and Ginkgo biloba extract and its active components.

Topics: Alkaloids; Animals; Benzofurans; Berberine; China; Cholinesterase Inhibitors; Diterpenes; Dopamine Agonists; Dopamine Antagonists; Drugs, Chinese Herbal; Epoxy Compounds; Ginkgo biloba; Humans; Immunosuppressive Agents; Lactams; Lignans; Neuroprotective Agents; Phenanthrenes; Rutaceae; Sesquiterpenes

Topics: Amyotrophic Lateral Sclerosis; Benzofurans; Double-Blind Method; Humans; Treatment Outcome

As one of the leading causes of morbidity and mortality, stroke and its recurrence has attracted more and more attention. Dl-3-n-butylphthalidle(NBP) has been widely used for treating acute ischemic stroke in China and shows a great clinical effect. NBP plays a role in different pathophysiological processes in the treatment of ischemic stroke, including antioxidants, anti-inflammatory, anti-apoptotic, anti-thrombosis, and mitochondrial protection. Many randomized, double-blind, placebo-controlled, multicenter clinical trials suggest that NBP is a safe and effective treatment for ischemic stroke. To sum up, the current research is mainly focused on the short-term treatment of stroke patients with RCT (randomized controlled trial). Therefore, we designed this study to confirm the role of butylphthalide in secondary stroke prevention in the real world.. This study will be a multicenter, prospective real-world trial. We would recruit 8000 patients with ischemic stroke from 78 public hospitals in China. All participants will be allocated to one of two parallel treatment groups according to their own wills: (1) butylphthalide group: 0.2 g of butylphthalide capsules three times daily plus routine treatment (aspirin 50-300 mg/d, clopidogrel 75 mg/d, etc.); (2) control group: routine treatment (aspirin 50-300 mg/d, clopidogrel 75 mg/d, etc.). Treatment duration is 90 consecutive days or more. The primary outcome is recurrence rate of stroke within 1 month, 3 months, 6 months and 1 year in butylphthalide group and control group. The secondary outcomes included NIHSS score, the mRS score, other clinical cardiovascular events within one year (sudden death / myocardial infarction / arrhythmia / heart failure, etc.), and adverse events of patients in groups. NIHSS will be captured in the first month after discharge, and the others will be captured at the same time points as the primary end point.. This trial will be exploring the efficacy and safety of butylphthalide in secondary prevention of ischemic stroke to expand the scope of application of butylphthalide soft capsules and provide new ideas for enriching the secondary prevention of stroke.. Chinese Clinical Trial Registry (ChiCTR).. ChiCTR2000034481. Registered on 6 July 2020, http://www.chictr.org.cn/showproj.aspx?proj=55800.

Topics: Aspirin; Benzofurans; Clopidogrel; Double-Blind Method; Humans; Internet; Ischemic Stroke; Multicenter Studies as Topic; Prospective Studies; Randomized Controlled Trials as Topic; Secondary Prevention; Treatment Outcome

Stress is not scarce in peoples' daily life that may result in mental diseases and cognitive impairments. Chronic restraint stress (CRS) is a well-validated animal model used to investigate the mechanism of stress-associated depression and cognitive impairments. Dl-3-n-butylphthalide (NBP) possesses anti-oxidant, anti-inflammatory and anti-apoptotic, promoting neurogenesis and neuroplasticity that exerts neuroprotective effects. However, the effects of NBP on CRS-induced depression and cognitive impairments remain unclear.. C57BL/6 male mice were randomly divided into the control group, stress group and stress+NBP group. Mice were exposed to CRS for three consecutive weeks and mice in the NBP treatment group were administered with NBP before the CRS procedure. After that, depression and cognition behaviors were evaluated followed by phosphorylation of Ca2+/calmodulin-dependent protein kinase II (p-CaMKII), phosphorylation of cAMP-response element-binding protein (p-CREB), brain-derived neurotrophic factor (BDNF) proteins expression, immunohistochemistry of hippocampal postsynaptic density 95 (PSD95) and synaptophysin, and hippocampal morphology.. Our results showed that mice exhibited depression-like behaviors and cognitive deficits after 3 weeks exposure to CRS. Additionally, CRS downregulated CaMKII/CREB/BDNF signaling pathway, reduced PSD95 and synaptophysin expression and induced hippocampal CA1 and dentate gyrus ment significantly reversed the hippocampal pathological and molecular changes induced by CRS.. In conclusion, these results reveal that NBP exerts a neuroprotective effect on depression and cognitive deficit through activating CaMKII/CREB/BDNF pathway, enhancing PSD95 and synaptophysin expression and protecting hippocampal morphology.

Topics: Animals; Benzofurans; Brain-Derived Neurotrophic Factor; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cognitive Dysfunction; Cyclic AMP Response Element-Binding Protein; Depression; Hippocampus; Male; Mice; Mice, Inbred C57BL; Neuroprotective Agents; Signal Transduction; Synaptophysin

Stroke can cause physical and mental problems. This study examined how the sequential therapy of N-butylphthalide (NBP) could effectively improve physical movement, life activities, and psychological disorders in stroke patients.. This double-blind, randomized controlled trial included middle-aged or elderly patients with acute ischemic stroke that had commenced within 48 hours before enrolment in the study. The experimental group was administered 100 mL NBP injections twice a day in the first 14 days, and a sequential 200 mg NBP soft capsule 3 times a day for the next 76 days. The control group was administered 100 mL NBP placebo injections twice a day in the first 14 days and 200 mg sequential NBP placebo soft capsule 3 times a day for the next 76 days. Primary outcomes were the National Institutes of Health Stroke Scale, the Barthel Index of activities of daily living, and Modified Rankin Scale which were evaluated at day 0, day 14, and month 1 or at day 14, month 3, and month 6. Secondary outcomes included the Hamilton Anxiety Scale and the Hamilton Depression Scale, all were evaluated on day 0, month 3, and month 6. Moreover, the adverse reaction of NBP or other serious adverse events were evaluated at each time.. Our therapy significantly increased the Barthel Index of activities of daily living scores, decreased the National Institutes of Health Stroke Scale and Modified Rankin Scale scores, and the incidence of the Hamilton Anxiety Scale and the Hamilton Depression Scale of ischemic stroke patients (P < .05).. Our results indicated that 90 days' sequential therapy with NBP as an additional therapy in the treatment of ischemic stroke can better improve patients' psychological and behavioral functions without significant side effects.

Topics: Activities of Daily Living; Adult; Aged; Aged, 80 and over; Behavioral Symptoms; Benzofurans; Double-Blind Method; Female; Humans; Ischemic Stroke; Male; Mental Disorders; Middle Aged; Treatment Outcome

As a neuroprotective medication, butylphthalide (NBP) may help protect against cerebral ischaemic injury. However, evidence on whether NBP influences the outcomes of patients who had acute ischaemic stroke who are receiving revascularisation treatment is limited. This study aims to evaluate whether additional NBP therapy can improve the functional outcome of patients who receive intravenous recombinant tissue plasminogen activator and/or endovascular treatment (EVT).. The study will be a randomised, double-blind, placebo-controlled, multiple-centre, parallel group trial. The sample size is estimated at 1200 patients. Eligible patients will be randomised at a 1:1 ratio to receive either NBP or placebo daily for 90 days, which will include 14 days of injections and 76 days of capsules. The first use of NBP/placebo will be started within 6 hours of onset of ischaemic stroke. The primary outcome is the functional outcome as assessed by the 90-day modified Rankin Scale, adjusted for baseline scores on the National Institutes of Health Stroke Scale. The primary safety outcome is the percentage of serious adverse events during the 90 days of treatment. This trial will determine whether NBP medication benefits patients who had acute ischaemic stroke who receive intravenous thrombolysis or EVT.. The protocol was written according to the general ethical guidelines of the Declaration of Helsinki and approved by the Institutional Review Board/Ethics Committee of Beijing Tiantan Hospital, Capital Medical University with approval number KY 2018-003-02. Ethics committees of all participating sites have approved the study . Results of the study will be published in peer-reviewed scientific journals and shared in scientific presentations.. NCT03539445.

Topics: Benzofurans; Brain Ischemia; Double-Blind Method; Humans; Ischemic Stroke; Randomized Controlled Trials as Topic; Stroke; Thrombolytic Therapy; Tissue Plasminogen Activator; Treatment Outcome

Topics: Adult; Aged; Aged, 80 and over; Benzofurans; Brain Infarction; Diffusion Magnetic Resonance Imaging; Female; Humans; Injections; Ischemic Attack, Transient; Male; Middle Aged; Neuroprotective Agents; Outcome Assessment, Health Care; Severity of Illness Index

To determinate the clinical effect of butyphthalide combined with idebenone in the treatment of vascular dementia (VD) and the influence on inflammatory cytokines and vascular endothelial functions.. Clinical comparative study.. Department of Neurology, Baoding First Central Hospital, from June 2017 to June 2018.. Eighty-eight VD patients were divided into observation group (44 cases) and control group (44 cases) at random. Idebenone was given to the control group, and butyphthalide combined with idebenone was given to the observation group for 12 weeks. C-reactive protein (CRP), tumor necrosis factor α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were detected before and after the treatment to evaluate the level of serum inflammatory factors. Peripheral blood endothelial microparticles (EMPs), endothelin (ET-1), and vascular endothelial growth factor (VEGF) were detected to evaluate vascular endothelial functions. Mini-mental state examination (MMSE), clinical dementia scale (CDR), and ability of daily life (ADL), were used to evaluate cognitive function, dementia degree, and self-care ability in daily life. The occurrences of adverse reactions were recorded.. Before the treatment, the comparison differences in the indexes of both groups had no statistical significance (p>0.05). After the treatment, the scores of CD62E+, VEGF, and MMSE of observation group rose obviously, compared with those before the treatment, and were significantly higher than those of control group (p <0.05). After the treatment, the scores of IL-6, CRP, TNF-α, IL-1β, CD31+, CDl44+, ET-1, CDR and ADL of observation group significantly lowered, compared with those before the treatment, and were significantly lower than those of control group (p <0.05). The differences in the adverse reactions of both groups had no statistical significance (p >0.05).. Butyphthalide combined with idebenone can effectively reduce serum inflammatory factor level of VD patients, regulate vascular endothelial functions, relieve dementia degree, and improve cognitive function and daily activity ability.

Topics: Activities of Daily Living; Aged; Antioxidants; Benzofurans; C-Reactive Protein; Cytokines; Dementia, Vascular; Drug Therapy, Combination; Endothelin-1; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Ubiquinone; Vascular Endothelial Growth Factor A

DL-3-n-butylphthalide (NBP) was demonstrated to increase the cerebral blood flow (CBF) in the animal models, but there are no clinic studies to verify this. We aimed to explore the effect of NBP on improving cerebral hypoperfusion caused by cerebral large-vessel stenosis.. In this single-center, randomized, double-blind, placebo-controlled study, 120 patients with severe carotid atherosclerotic stenosis and cerebral hypoperfusion in the ipsilateral middle cerebral artery (MCA) were included and randomly assigned into NBP or placebo group as 1:1 radio. Patients in NBP or placebo group received 200 mg or 20 mg of NBP capsules three times daily for four weeks respectively. Single photon emission computed tomography (SPECT) was used to assess regional CBF (rCBF) in four regions of interest (ROIs) corresponding to MCA before and 12 weeks after the treatment. After therapy, the rCBF change for every ROI and the whole CBF change in MCA territory for every patient were classified into amelioration, stabilization and deterioration respectively.. 48 NBP patients (6 with bilateral stenosis) and 46 placebo patients (8 with bilateral stenosis) completed the trial. Overall, both groups had 54 stenotic carotid arteries and 216 ROIs for rCBF change analysis. After therapy, the rCBF in ROIs increased in NBP group (83.5% ± 11.4% vs. 85.8% ± 12.5%, p = 0.000), whereas no change was found in placebo group (86.9% ± 11.6% vs. 87.8% ± 11.7%, p = 0.331). Besides, there was higher percentages of ROIs with rCBF amelioration and stabilization in NBP group than in placebo group (93.1% vs. 79.2%, p = 0.000). Furthermore, ordinal regression analysis showed that compared with placebo, NBP independently made more patients to have whole CBF amelioration in ipsilateral MCA (Wald-χ2 = 5.247, OR = 3.31, p = 0.022).. NBP might improve the cerebral hypoperfusion in the patients with carotid artery atherosclerotic stenosis.. Chinese Clinical Trial Registry, ChiCTR1900028005, registered December 8th 2019- Retrospectively registered (http://www.chictr.org.cn/index.aspx).

Topics: Benzofurans; Double-Blind Method; Humans; Intracranial Arteriosclerosis; Neuroprotective Agents

The effectiveness of neuroprotective agents is still unclear. Here we analyzed the clinical outcomes of acute ischemic stroke (AIS) patients treated with human urinary kallidinogenase (HUK) or edaravone (Eda) combined with butylphthalide (NBP).. From January 2016 to December 2017, a total of 165 AIS patients were enrolled in this open-label, randomized controlled clinical study. Patients were randomly allocated into HUK group and Eda group in a ratio of 2:1. All the patients received basic treatments and NBP (200 mg p.o. qid) while HUK group received 0.15 PNA unit of HUK injection (ivgtt. qd) and Eda group received 30 mg Eda (ivgtt. bid) for 14 consecutive days. Independency rate [12-month modified Rankin Scale (mRS) score ≤ 1] and related factors were compared between the two groups.. Twelve-month mRS score of the HUK group (1, IQR 0~1) was significantly lower compared with Eda group (2, IQR 1~3, p < .0001). The HUK treatment achieved an independency rate of 79.1% while the Eda treatment only had 45.3% (p < .0001). Further binary logistic regression showed that recurrent stroke (RR: 0.1, 95% CI: 0.0~0.1, p = .038) and HUK treatment (RR: 4.2, 95% CI: 1.1~16.5, p = .041) could significantly affect patients' 12-month outcomes.. Human urinary kallidinogenase combined with NBP can enhance AIS patients' long-term independency rate, and the effectiveness of HUK combined therapy is better than Eda.

Topics: Adult; Aged; Aged, 80 and over; Benzofurans; Brain Ischemia; Drug Therapy, Combination; Edaravone; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Stroke; Tissue Kallikreins; Treatment Outcome; Young Adult

This study aims to explore the curative effect of dl-3-n-butylphthalide (NBP) on patients with acute cerebral infarction (ACI) and its effects on serum lipoprotein-associated phospholipase A2 (Lp-PLA2) and hypersensitive C-reactive protein (hs-CRP) levels.. A total of 136 ACI patients treated in our hospital, who met the criteria, were selected and randomly divided into two groups: control group (n = 60, including 28 males and 32 females) and treatment group (n = 76, including 32 males and 44 females). Patients in the control group were treated with routine drug therapy, while patients in the treatment group were treated with NBP on this basis. A dose of 100 ml was administered by intravenous injection for 2 times/day, for 14 days. The curative effect was evaluated using the National Institute of Health Stroke Scale (NIHSS) and Barthel index (BI) self-care ability. The levels of the two factors in serum were measured using enzyme-linked immunosorbent assay, and the changes in levels of these two factors in serum at different time points before and after treatment were compared between the two groups.. (a) Lp-PLA2 and hs-CRP levels in the treatment group after treatment were significantly lower than those before treatment and those in the control group after treatment (p < .05). (b) The NIHSS and BI scores in the treatment group were significantly lower after treatment than before treatment and those in the control group after treatment (p < .05).. Dl-3-n-butylphthalide can improve the expression of Lp-PLA2 and hs-CRP in serum in ACI patients. Furthermore, NBP has significant efficacy in inhibiting inflammation and improving neurological symptoms.

Topics: 1-Alkyl-2-acetylglycerophosphocholine Esterase; Acute Disease; Benzofurans; C-Reactive Protein; Cerebral Infarction; Drug Monitoring; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Treatment Outcome

In China, both human urinary kallindinogenase (HUK) and 3-n-butylphthalide (NBP) are recommended for clinical use to improve cerebral blood circulation during an acute ischemic stroke (AIS). The objective was to evaluate the economic value of HUK vs NBP for patients with AIS from a Chinese payer's perspective.. An economic evaluation based on data of patients who have been treated with either HUK (n = 488) or NBP (n = 885) from a prospective, phase IV, multi-center, clinical registry study (Chinese Acute Ischemic Stroke Treatment Outcome Registry, CASTOR) was conducted to analyze the cost and effectiveness of HUK vs NBP for AIS in China. Before the analysis, the patients were matched using propensity score. Both a cost-minimization analysis and a cost-effectiveness analysis were conducted to compare the matched pairs. A bootstrapping exercise was conducted for the matched arms to demonstrate the probability of one intervention being cost-effective over another for a given willingness-to-pay for an extra quality-adjusted life-year (QALY).. After propensity score matching, 463 pairs were matched. The overall medical cost in the HUK arm is USD 2,701.20, while the NBP arm is USD 3,436.83, indicating HUK is preferred with cost-minimization analysis. Although the QALY gained in the HUK arm (0.77176) compared with the NBP arm (0.76831) is statistically insignificant (p = .4862), the cost-effectiveness analysis as exploratory analysis found that, compared with NBP, HUK is a cost-saving strategy with the lower costs of USD 735.63 and greater QALYs gained of 0.00345. Among the 5,000 bootstrapping replications, 100% indicates that HUK is cost-effective compared with NBP under a 1-time-GDP threshold; and 97.12% indicates the same under a 3-time-GDP threshold.. This economic evaluation study indicates that administrating HUK is a cost-saving therapy compared with NBP for managing blood flow during AIS in the Chinese setting.

Topics: Benzofurans; Cerebrovascular Circulation; China; Cost-Benefit Analysis; Female; Health Services; Humans; Male; Models, Econometric; Prospective Studies; Quality-Adjusted Life Years; Stroke

BACKGROUND Delayed encephalopathy after acute carbon monoxide (CO) poisoning (DEACMP) is one of the most serious complications after CO poisoning. This study was conducted to explore the efficacy of the combined application of N-Butylphthalide and hyperbaric oxygenation therapy (HBO) on cognitive dysfunction in patients with DEACMP. MATERIAL AND METHODS A total of 184 patients with DEACMP were randomly assigned to either receive HBO or N-Butylphthalide and HBO. Meanwhile, all patients received conventional treatment. The total remission rate (RR) was used to assess the clinical efficacy. The Mini-Mental State Examination (MMSE) was used to assess the cognitive function, and the National Institutes of Health Stroke Scale (NIHSS) was used to assess the neurological function. RESULTS Finally, there were 90 and 94 patients in the control and experimental groups, respectively. After eight weeks of treatment, the total RR in the experimental group (47.9%) was significantly higher than that in the control group (33.3%). Compared to the control group, significantly more patients in the experimental group had MMSE scores of 24-30. The lower NIHSS score in the experimental group showed that N-Butylphthalide had the effect of preservation and restoration of neurological function. No obvious drug toxicity or liver and kidney dysfunction was observed, and there was no significant change in the level of blood glucose and blood lipids. CONCLUSIONS These results indicated that the combined application of N-Butylphthalide and HBO could significantly improve the cognitive dysfunction of patients with DEACMP and have great clinical efficacy, which should be further studied.

Topics: Acute Disease; Benzofurans; Brain Diseases; Carbon Monoxide Poisoning; Cognitive Dysfunction; Demography; Female; Humans; Hyperbaric Oxygenation; Male; Middle Aged; Neuropsychological Tests; Remission Induction; Treatment Outcome

Progressive cerebral infarction (PCI) is associated with high rates of mortality and disability. Many studies have shown that Dl-3n-butylphthalide (NBP) is effective against acute ischemic stroke. The administration of NBP can result in an increased number of capillaries in the ischemic region, promote the establishment of collateral circulation, protect the mitochondria, and narrow the infarction area, among other effects. In the present study, we evaluated the efficacy and safety of NBP for the treatment of PCI.Between March 2008 and May 2012, we performed a randomized, double-blind placebo-controlled study including 304 inpatients with PCI. These patients were randomly assigned to the test (152 cases) and control groups (152 cases). The test group received 200 mg of NBP soft capsules orally, 15 minutes before each meal, 3 times daily. The control group received 200 mg of placebo soft capsules orally, 15 minutes before each meal, 3 times daily. Treatment was administered during 21 days. The National Institute of Health Stroke Scale (NIHSS) score was assessed before the treatment and on days 7, 14, 21, and 30 after treatment. The Barthel index (BI) was assessed on the same days and on day 90.In the test group, the NIHSS scores on days 7, 14, 21, and 30 were 14.75 ± 4.85, 11.62 ± 3.49, 8.87 ± 5.17, and 6.38 ± 4.93, respectively. In the control group, they were 16.08 ± 3.76, 13.28 ± 5.02, 11.05 ± 4.25, and 8.43 ± 5.41 (P < .05), respectively. The BI on days 7, 14, 21, 30, and 90 were 51.57 ± 15.11, 61.21 ± 16.39, 70.48 ± 18.21, 76.41 ± 19.02, and 81.10 ± 15.52 for the test group and 46.79 ± 18.42, 55.93 ± 19.12, 64.84 ± 17.67, 70.65 ± 18.54, and 76.54 ± 17.05 for the control group (P < .05), respectively. Adverse events were elevation of alanine aminotransferase and aspartate aminotransferase (P > .05).NBP was useful to improve the outcome of patients with PCI and decreased their disability for activities of daily living. NBP was an efficacious and safe treatment for PCI.

Topics: Activities of Daily Living; Administration, Oral; Adult; Aged; Alanine Transaminase; Aspartate Aminotransferases; Benzofurans; Cerebral Infarction; Disability Evaluation; Disease Progression; Double-Blind Method; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Severity of Illness Index; Time Factors; Treatment Outcome

To observe the curative effect of dl-3-n-Butylphthalide (NBP) on patients with acute cerebral infarction (ACI) and its effects on levels of serum vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).. A total of 160 ACI patients treated in our hospital who met the criteria were selected and randomly divided into treatment group (n=80, including 42 males and 38 females) and control group (n=80, including 40 males and 40 females). The control group was treated with routine drug therapy, while the treatment group was treated with butylphthalide on this basis. The curative effect was evaluated using the National Institute of Health Stroke Scale (NIHSS) and the Activity of Daily Life Scale (ADL Scale). The levels of the two factors in serum were measured using enzyme-linked immunosorbent assay (ELISA), and the changes in the levels of the two factors in serum at different time points before and after treatment were compared between the two groups.. After treatment, the levels of the two factors in serum in both groups were significantly increased compared with those before treatment (p<0.05), and the increase in treatment group was more significant than that in control group (p<0.05). The scores of ADL scale in both groups were significantly increased after treatment compared with those before treatment, and the increase in treatment group was more significant than that in control group (p<0.05). The scores of NIHSS in both groups were significantly decreased compared with those before treatment, and the decrease in treatment group was more significant than that in control group (p<0.05).. NBP can improve the expressions of VEGF and bFGF in serum of ACI patients, and its effect is superior to that of conventional drugs.

Topics: Activities of Daily Living; Adult; Aged; Aged, 80 and over; Benzofurans; Cerebral Infarction; Female; Fibroblast Growth Factor 2; Humans; Male; Middle Aged; Neuroprotective Agents; Stroke; Vascular Endothelial Growth Factor A; Young Adult

Vascular cognitive impairment without dementia is very common among the aged and tends to progress to dementia, but there have been no proper large-scale intervention trials dedicated to it. Vascular cognitive impairment without dementia caused by subcortical ischemic small vessel disease (hereinafter, subcortical Vascular cognitive impairment without dementia) represents a relatively homogeneous disease process and is a suitable target for therapeutic trials investigating Vascular cognitive impairment without dementia. Preclinical trials showed that dl-3-n-butylphthalide (NBP) is effective for cognitive impairment of vascular origin.. In this randomized, double-blind, placebo-controlled trial, we enrolled patients aged 50-70 years who had a diagnosis of subcortical Vascular cognitive impairment without dementia at 15 academic medical centers in China. Inclusion criteria included a clinical dementia rating ≥0.5 on at least one domain and global score ≤0.5; a mini-mental state examination score ≥20 (primary school) or ≥24 (junior school or above); and brain magnetic resonance imaging consistent with subcortical ischemic small vessel disease. Patients were randomly assigned to NBP 200 mg three times daily or matched placebo (1:1) for 24 weeks according to a computer-generated randomization protocol. All patients and study personnel were masked to treatment assignment. Primary outcome measures were the changes in Alzheimer's disease assessment scale-cognitive subscale (ADAS-cog) and clinician's interview-based impression of change plus caregiver input (CIBIC-plus) after 24 weeks. All patients were monitored for adverse events (AEs). Outcome measures were analyzed for both the intention-to-treat (ITT) population and the per protocol population.. This study enrolled 281 patients. NBP showed greater effects than placebo on ADAS-cog (NBP change -2.46 vs. placebo -1.39; P = .03; ITT) and CIBIC-plus (80 [57.1%] vs. 59 [42.1%] patients improved; P = .01; ITT). NBP-related AE were uncommon and primarily consisted of mild gastrointestinal symptoms.. Over the 6-month treatment period, NBP was effective for improving cognitive and global functioning in patients with subcortical vascular cognitive impairment without dementia and exhibited good safety.

Topics: Aged; Benzofurans; Cerebral Small Vessel Diseases; China; Cognition Disorders; Double-Blind Method; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Neuropsychological Tests

Delayed encephalopathy after carbon monoxide (CO) poisoning (DEACMP) is still a clinical challenge. This study aimed to investigate the efficacy of combined therapy of mesenchymal stem cell (MSC) transplantation and butylphthalide in DEACMP patients.Forty-two DEACMP patients were treated with 1 of the 3 therapies: combined therapy of MSC transplantation and butylphthalide; MSC transplantation alone; or hyperbaric oxygen therapy. The MSCs were alternatively injected into the subarachnoid space and the carotid artery using a self-made high-pressure injector. The Mini-Mental State Examination and the Barthel index of activities of daily living were administered before the treatment, and at 1 month, 3 months, and 6 months after the treatment. Computed tomography and magnetic resonance imaging results before and after the treatment were compared.At 1 month, 3 months, and 6 months after the treatment, the Mini-Mental State Examination scores and the Barthl scores were significantly higher in patients with the combined therapy of MSC transplantation and butylphthalide than those in patients with MSC transplantation alone or hyperbaric oxygen therapy (all P < 0.0001). No significant adverse events occurred.The combination of MSC transplantation and butylphthalide is safe and effective in treating DEACMP.

Topics: Adult; Analysis of Variance; Benzofurans; Brain Diseases; Carbon Monoxide Poisoning; Combined Modality Therapy; Female; Follow-Up Studies; Humans; Magnetic Resonance Imaging; Male; Mesenchymal Stem Cell Transplantation; Middle Aged; Prospective Studies; Reference Values; Risk Assessment; Severity of Illness Index; Survival Rate; Tomography, X-Ray Computed; Treatment Outcome

To investigate the effect of Butylphthalide and Xue Shuan Tong on serum inflammatory factors and prognosis of patients with cerebral infarction. One hundred and twenty patients with acute cerebral infarction were randomly divided into control group, Butylphthalide group and Xue Shuan Tong group, with 40 patients in each group. Conventional therapy was performed in the control group; On the basis of conventional therapy, 100ml Butylphthalide intravenously twice a day was administrated among patients in Butylphthalide group; On the basis of conventional therapy, 250ml 0.9% NaCl intravenously once a day was conducted among patients in Xue Shuan Tong group. A treatment course of continuous 7 days was taken in the three groups. The serum levels of IL-2 and CGRP were detected for patients in the three groups before and after treatment. Carotid plaque thickness and size as well as intima-media thickness were detected by ultrasonic testing for patients in three groups before treatment and 90 days after follow-up. The NIHSS, Barthel and MRS scoring were performed for all the patients after 90-day follow-up to evaluate the prognosis. After treatment, differences in the levels of IL-2 and CGRP for patients in the three groups showed statistical significance (P<0.05), while the levels of IL-2 and CGRP in Xue Shuan Tong group were significantly higher than those in the other two groups (P<0.05). After 7-day treatment, plaque size and thickness in Xue Shuan Tong group and Butylphthalide group were significantly reduced, compared with those before treatment (P<0.05), but no significant differences was shown in the plaque size and thickness between Xue Shuan Tong group and Butylphthalide group (P>0.05) .The CA-IMT in Xue Shuan Tong group and Butylphthalide group was significantly reduced after treatment, and that in Butylphthalide group was significantly larger than that in Xue Shuan Tong group (P<0.05). After 90-day follow-up, NIHSS scores in Butylphthalide group were significantly less than those in the other two groups (P<0.05). After 90-day follow-up, Barthel scores in Butylphthalide group and Xue Shuan Tong group were significantly larger than those in control group (P<0.05), while differences between Butylphthalide group and Xue Shuan Tong group indicated no statistical significance (P>0.05). There were significant differences in MRS scores among patients in the three groups after 90-day follow-up (P<0.05). Butylphthalide and Xue Shuan Tong are clinically effective

Topics: Aged; Aged, 80 and over; Benzofurans; Calcitonin Gene-Related Peptide; Carotid Arteries; Carotid Intima-Media Thickness; Cerebral Infarction; Drugs, Chinese Herbal; Female; Humans; Inflammation; Interleukin-2; Male; Middle Aged; Nervous System Diseases; Plaque, Atherosclerotic; Prognosis

Dl-3-n-butylphthalide (NBP), first isolated from the seeds of celery, showed efficacy in animal models of stroke. This study was a clinical trial to assess the efficacy and safety of NBP with a continuous dose regimen among patients with acute ischemic stroke.. A randomized, double-blind, double-dummy trial enrolled 573 patients within 48 hours of onset of ischemic stroke in China. Patients were randomly assigned to receive a 14-day infusion of NBP followed by an NBP capsule, a 14-day infusion of NBP followed by aspirin, or a 14-day infusion of ozagrel followed by aspirin. The efficacy measures were Barthel index score and the modified Rankin scale (mRS) at day 90. Differences among the three groups on mRS were compared using χ(2) test of proportions (with two-sided α = 0.05) and Logistic regression analysis was conducted to take the baseline National Institutes of Health Stroke Scale (NIHSS) score into consideration.. Among the 535 subjects included in the efficacy analysis, 90-day treatment with NBP was associated with a significantly favorable outcome than 14-day treatment with ozagrel as measured by mRS (P < 0.001). No significant difference was found among the three groups on Barthel index at day 90. The rate of adverse events was similar among the three groups.. The 90-day treatment with NBP could improve outcomes at the third month after stroke. The NBP treatment (both intravenous and oral) is safe (ChiCTR-TRC-09000483).

Topics: Adult; Aged; Benzofurans; Double-Blind Method; Female; Humans; Male; Middle Aged; Stroke; Treatment Outcome

3-n-Butylphthalide (NBP) is a cardiovascular drug currently used for the treatment of cerebral ischemia. The present study aims to investigate the metabolism, pharmacokinetics, and excretion of NBP in humans and identify the enzymes responsible for the formation of major metabolites. NBP underwent extensive metabolism after an oral administration of 200 mg NBP and 23 metabolites were identified in human plasma and urine. Principal metabolic pathways included hydroxylation on alkyl side chain, particularly at 3-, ω-1-, and ω-carbons, and further oxidation and conjugation. Approximately 81.6% of the dose was recovered in urine, mainly as NBP-11-oic acid (M5-2) and glucuronide conjugates of M5-2 and mono-hydroxylated products. 10-Keto-NBP (M2), 3-hydroxy-NBP (M3-1), 10-hydroxy-NBP (M3-2), and M5-2 were the major circulating metabolites, wherein the areas under the curve values were 1.6-, 2.9-, 10.3-, and 4.1-fold higher than that of NBP. Reference standards of these four metabolites were obtained through microbial biotransformation by Cunninghamella blakesleana. In vitro phenotyping studies demonstrated that multiple cytochrome P450 (P450) isoforms, especially CYP3A4, 2E1, and 1A2, were involved in the formation of M3-1, M3-2, and 11-hydroxy-NBP. Using M3-2 and 11-hydroxy-NBP as substrates, human subcellular fractions experiments revealed that P450, alcohol dehydrogenase, and aldehyde dehydrogenase catalyzed the generation of M2 and M5-2. Formation of M5-2 was much faster than that of M2, and M5-2 can undergo β-oxidation to yield phthalide-3-acetic acid in rat liver homogenate. Overall, our study demonstrated that NBP was well absorbed and extensively metabolized by multiple enzymes to various metabolites prior to urinary excretion.

Topics: Administration, Oral; Adult; Alcohol Dehydrogenase; Animals; Benzofurans; Biotransformation; Cardiovascular Agents; Chromatography, Liquid; Cunninghamella; Cytochrome P-450 Enzyme System; Humans; Hydroxylation; Intestinal Absorption; Isoenzymes; Kidney; Liver; Male; Metabolomics; Molecular Structure; Oxidation-Reduction; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Electrospray Ionization; Substrate Specificity; Young Adult

To investigate the clinical effect of butylphthalide combined with urinary kallidinogenase in the treatment of chronic cerebral circulatory insufficiency (CCCI).. A total of 102 CCCI patients admitted to our hospital from October 2020 to December 2021 were retrospectively enrolled in this study. According to the different therapeutic strategy, the patients were divided into combined group (treated with butylphthalide combined with urinary kallidinogenase, n = 51) and butylphthalide group (treated with butylphthalide, n = 51). Blood flow velocity and cerebral blood flow perfusion before and after treatment between the two groups were compared. The clinical efficacy and adverse events of the two groups were analyzed.. After treatment, the effective rate of the combined group was significantly higher than the butylphthalide group (p = .015). Before treatment, the blood flow velocity of middle cerebral artery (MCA), vertebral artery (VA), basilar artery (BA) were comparable (p > .05, respectively), while after treatment, the blood flow velocity of MCA, VA, and BA in combined group were faster than those in butylphthalide group (p < .001, respectively). Before treatment, the relative cerebral blood flow (rCBF), relative cerebral blood volume (rCBV), relative mean transmit time (rMTT) of the two groups were comparable (p > .05, respectively). After treatment, rCBF and rCBV in combined group were higher than those in butylphthalide group (p < .001, respectively), and rMTT in combined group was lower than that in butylphthalide group (p = .001). The rate of adverse events in the two groups were comparable (p = .558).. Butylphthalide combined with urinary kallidinogenase can improve the clinical symptoms of CCCI patients, and the effect is promising, which is worthy of clinical application.

Topics: Benzofurans; Blood Flow Velocity; Cerebrovascular Circulation; Humans; Platelet Aggregation Inhibitors; Retrospective Studies; Tissue Kallikreins; Treatment Outcome

To investigate the efficacy of butylphthalide combined with edaravone in the treatment of acute ischemic stroke and the effect on serum inflammatory factors.. One hundred and sixty patients with acute ischemic stroke who attended the neurovascular intervention department of our hospital from May 2020 to June 2022 were enrolled as study subjects for prospective analysis and were equally divided into a control group and an experimental group using the random number table method, with 80 cases in each group. The control group was treated with edaravone injection, while the experimental group was treated with butylphthalide combined with edaravone. The disease was recorded to compare the efficacy, erythrocyte sedimentation rate, homocysteine, serum inflammatory factors including tumor necrosis factor-. The total effective rate of treatment in the experimental group was 90.0% (72/80), while that of the control group was 62.5% (50/80), the total effective rate of the experimental group was significantly higher than that of the control group, and the difference was statistically significant (. The treatment of acute ischemic stroke with butylphthalide combined with edaravone has positive significance in improving blood circulation regulation and serum inflammatory factor levels and is reliable and worthy of clinical promotion.

Topics: Benzofurans; Edaravone; Humans; Inflammation; Interleukin-6; Ischemic Stroke; Stroke

Topics: Benzofurans; Brain; Dose-Response Relationship, Drug; Drug Discovery; Humans; Molecular Structure; Neuroprotective Agents; Receptors, N-Methyl-D-Aspartate; Stroke; Structure-Activity Relationship

To assess the potential effect of dl-3-N-butylphthalide (dl-NBP) for the proliferation and differentiation of neural stem cells (NSCs) against hypoxia and the underlying mechanism.. Hippocampal NSCs were obtained from fetal rats. NSCs combined with dl-NBP and single NSCs were cultured. The impact of siRNA-mediated hypoxia-inducible factor-1alpha (HIF-1α) knockdown on NSCs was detected with western blotting (WB) and quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR). Cell-counting kit-8 assay was used for evaluating the viability of NSCs. Levels of HIF-1α protein were measured using WB, and vascular endothelial growth factor (VEGF) expression was quantified using RT-qPCR and enzyme-linked immunosorbent assay.. Compared with 7 different concentrations of dl-NBP, 0.25 g/L was determined as the optimal concentration to significantly increase the viability of NSCs (p < 0.001). Dl-NBP can significantly increase the viability of hypoxic NSCs (p < 0.001) and improve the differentiation of hypoxic NSCs into astrocytes (p = 0.001) and oligodendrocytes (p < 0.001). Meanwhile, Dl-NBP can significantly elevate levels of HIF-1α protein (p < 0.001) and VEGF mRNA (p = 0.001) / protein (p < 0.001) in NSCs in the hypoxic environment. However, after transfection with HIF-1α siRNA in NSCs, the viability and differentiation of NSCs was not recovered using dl-NBP under the hypoxic condition, as well as levels of HIF-1α and VEGF.. Dl-NBP can reverse the weaker proliferation and differentiation power of NSCs in the hypoxic environment. The HIF-1α - VEGF pathway may be implicated in this protective effect of dl-NBP.

Topics: Animals; Benzofurans; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Neural Stem Cells; Neuroprotective Agents; Rats

Chronic cerebral ischemia is one of the common ischemic cerebrovascular diseases. Chronic cerebral ischemia can lead to brain dysfunction, and its pathophysiological mechanism involves inflammation, blood-brain barrier destruction, oxidative stress, and other factors. As it is difficult to detect, it is easily overlooked, and it is often only observed following the onset of cognitive dysfunction. At present, there are only a few drugs for its treatment. Dl-3-n-butylphthalide (NBP), a compound extracted from celery seed, may play an important role in protecting against brain damage caused by chronic cerebral ischemia. Therefore, here, we have paid attention to the prevention and treatment of chronic cerebral ischemia with NBP.

Topics: Benzofurans; Brain Injuries; Brain Ischemia; Humans; Neuroprotective Agents

A decrease in hippocampal neurogenesis is considered an important cause of cognitive impairment, while changes in mossy fiber sprouting are closely related to development of spontaneous recurrent seizures in chronic temporal lobe epilepsy (TLE). Racemic l-3-n-butylphthalide (DL-NBP) can alleviate cognitive impairment in ischemic stroke and Alzheimer's disease by promoting neurogenesis. DL-NBP treatment can also improve cognitive function and reduce seizure incidence in chronic epileptic mice. However, the mechanisms of action of DL-NBP remain unclear. The aim of the present study was to examine the effects of DL-NBP on mossy fiber sprouting, hippocampal neurogenesis, spontaneous epileptic seizures, and cognitive functioning in the chronic phase of TLE.. Nissl staining was used to evaluate hippocampal injury, while immunofluorescent staining was used to analyze hippocampal neurogenesis. The duration of spontaneous seizures was measured by electroencephalography. The Morris water maze was used to evaluate cognitive function. Timm staining was used to assess mossy fiber sprouting.. TLE animals showed reduced proliferation of newborn neurons, cognitive dysfunction, and spontaneous seizures. Treatment with DL-NBP after TLE increased the proliferation and survival of newborn neurons in the dentate gyrus, reversed the neural loss in the hippocampus, alleviated cognitive impairments, and decreased mossy fiber sprouting and long-term spontaneous seizure activity.. We provided pathophysiological and morphological evidence that DL-NBP might be a useful therapeutic for the treatment of TLE.

Topics: Animals; Benzofurans; Epilepsy, Temporal Lobe; Hippocampus; Mice; Mossy Fibers, Hippocampal; Neurogenesis; Rats

The goal of this study was to explore the mechanism of action of DL-3-n-butylphthalidein (NBP) the treatment of vascular dementia (VD) in mice. A vascular dementia mouse model was established with repeated cerebral ischemia/reperfusion (I/R), followed by administration of two different doses of NBP for 28 days. A Morris water maze was used to detect any changes in spatial cognition, while H&E staining was used to observe any histopathological changes in the hippocampus. The number of Caspase-3 and Caspase-9 positive neurons in the hippocampal CA1 region were also assessed using immunohistochemistry. The expression of Nrf2, Sirt3, and autophagy-related factors LC3 II/I and p62 in the hippocampus were detected by Western blotting. The results indicated that NBP treatment ameliorated learning and memory deficits, attenuated pathological damage in the CA1 regions, and reduced autophagy and apoptosis via the Nrf2/SIRT3 pathway after repeated cerebral I/R. Therefore, NBP treatment can improve the learning and cognitive memory of VD mice, possibly through the inhibition of autophagy and apoptosis mediated by the Nrf2/SIRT3 signaling pathway.

Topics: Animals; Apoptosis; Autophagy; Benzofurans; CA1 Region, Hippocampal; Dementia, Vascular; Disease Models, Animal; Mice; Neuroprotective Agents; NF-E2-Related Factor 2; Sirtuin 3

Cerebral venous disorder may have a harmful effect on ischaemic stroke; however, the underlying mechanism remains to be elucidated. Although Dl-3-n-butylphthalide is a multitarget agent for antiischaemic stroke, its neuroprotective role in brain ischaemia accompanied by brain venous disturbance remains unclear. In this study, we induced cerebral venous disturbance by the occlusion of bilateral external jugular veins (EJVs) to explore the potential mechanism of the adverse effects of cerebrovenous disorders in cerebral infarction and explore the protective effect of Dl-3-n-butylphthalide on cerebral infarction accompanied through cerebral venous disturbance.. Cerebral venous disturbance was induced in Sprague-Dawley rats through the permanent occlusion of bilateral EJVs, and cerebral ischaemic stroke was induced through the permanent occlusion of the right cortical branches of the middle cerebral artery. 2,3,5-triphenyltetrazolium chloride staining, MRI, Evans blue extravasation and behavioural test were performed to evaluate infarction volume, cerebral blood flow (CBF), blood-brain barrier (BBB) integrity and neurological function. Immunofluorescence staining and western blot analysis were performed to detect loss of neuron, endothelial cells, pericytes and tight junctions.. Bilateral EJVs occlusion did not cause cerebral infarction; however, it increased the infarction volume compared with the simple middle cerebral artery occlusion (MCAO) group, accompanied by severe neuron loss, worse neurological function, lower CBF, increased EJVs pressure, exacerbated Evans blue extravasation and brain oedema, as well as attenuated angiogenesis. Dl-3-n-butylphthalide displayed a neuroprotective effect in rats with MCAO accompanied by EJVs occlusion by reducing neuron loss, accelerating CBF restoration, promoting angiogenesis and relieving BBB damage.. Bilateral EJVs occlusion did not significantly affect normal rats but aggravated brain damage in the case of ischaemic stroke. Dl-3-n-butylphthalide treatment plays a neuroprotective role in rats with MCAO accompanied by EJVs occlusion, mainly due to the promotion of CBF restoration and BBB protection.

Topics: Animals; Benzofurans; Brain Injuries; Brain Ischemia; Drainage; Endothelial Cells; Evans Blue; Infarction, Middle Cerebral Artery; Ischemic Stroke; Rats; Rats, Sprague-Dawley; Stroke

To explore the effects of butylphthalide on the levels of serum CRP, PAPK7, NT-3 and neurological function in patients with acute cerebral infarction (ACI). 120 patients with ACI who were treated at Peking University First Hospital from September 2014 to June 2016 were selected as the research objects. The patients were randomly divided into a control group and an observation group, with 60 cases in each group. Conventional methods were adopted in the control group, and the observation group used butylphthalide for treatment. Two months later, the clinical efficacy, serum C-reactive protein (CRP), Parkinson's disease protein 7 (PAPK7), neurotrophic factor-3 (NT-3) levels, and the National Institutes of Health Stroke Scale (NIHSS) score before and after treatment were put into comparison and analysis. Before treatment, the NIHSS score showed no significant difference between the two groups (p>0.05); An observably higher NIHSS score of the observation group compared with the control group was seen after treatment (p=0.000). Butylphthalide has a significant therapeutic effect on patients with ACI. It can effectively restore the patients' neurological function, and remarkably improve the serum CRP, PAPK7 and NT-3 levels, which is worthy of clinical promotion.

Topics: Aged; Benzofurans; C-Reactive Protein; Cerebral Infarction; Female; Gene Expression Regulation; Humans; Male; Neuroprotective Agents; Neurotrophin 3; Protein Deglycase DJ-1

Topics: Animals; Atherosclerosis; Benzofurans; Brain Ischemia; Endothelial Cells; Mice; Neuroprotective Agents; Oxidative Stress; Rats; Stroke

Topics: Alzheimer Disease; Benzofurans; Humans; Neurodegenerative Diseases; Neuroprotective Agents

For investigating an influence on butylphthalide sodium chloride injection combined with edaravone dexborneol on neurological function and serum inflammatory factor levels in sufferers having acute ischemic stroke, 120 sufferers having acute ischemic stroke from September 2020 to September 2021 are chosen for the study subjects. In line with the diverse therapies, they took part in a control group and the study group, with 60 examples in each group. The control group is treated with edaravone dexborneol, and the study group is treated with butylphthalide sodium chloride injection, based on the control group. The posttreatment curative efficacy on the two groups is recorded, and treatment of both the two groups is compared. Before and after neurological function indexes (NIHSS and mRS), inflammatory factor indexes (IL-6, CRP, and TNF-

Topics: Benzofurans; Edaravone; Humans; Interleukin-6; Ischemic Stroke; Quality of Life; Sodium Chloride; Stroke; Treatment Outcome; Tumor Necrosis Factor-alpha

To assess cerebral hemodynamic changes by transcranial doppler ultrasound in patients with acute cerebral infarction before and after treatment with butylphthalide, A total of 90 patients with acute cerebral infarction admitted to our hospital from January 2019 to January 2020 were selected and equally divided into the control group and the experimental group according to the order of admission. The control group was treated with conventional treatment, while the experimental group was additionally given butylphthalide drug treatment. The experimental group obtained better hemodynamic indexes as compared with the control group (P<0.05). The experimental group yielded a notably higher total clinical effective rate after treatment in contrast with the control group (P<0.05). After treatment, the serum indexes of the experimental group were evidently lower than those of the control group (P<0.05). After treatment, a remarkably lower NIHSS score of the experimental group than the control group was observed (P<0.05). The BI index score of the experimental group after treatment was considerably higher than that of the control group (P<0.05). After treatment, the MMSE score in the experimental group was significantly higher than it was in the control group (P<0.05). The treatment of butylphthalide in patients with acute cerebral infarction can effectively improve the clinical symptoms of the patients and the cerebral hemodynamics of the patients tested by TCD found that this treatment yields an excellent therapeutic effect and is worthy of promotion and application.

Topics: Acute Disease; Benzofurans; Brain Ischemia; Cerebral Infarction; Hemodynamics; Humans; Stroke; Ultrasonography, Doppler, Transcranial

Topics: Animals; Benzofurans; Chemical and Drug Induced Liver Injury; Ischemia; Ischemic Stroke; Magnetic Resonance Spectroscopy; Neuroprotective Agents; Rats; Stroke

The treatment of acute ischemic stroke (AIS) remains a tough challenge in clinic. Here, we report the anti-AIS activity of R- and S-FMPB generated from hybridization of ring-opened R- and S-3-N-butylphthalide (R- and S-NBP) derivatives (R- and S-APB) with 4-fluro-edaravone (4-F-Eda), respectively. S-FMPB (10 mg/kg, iv) significantly improved the neurological score and alleviated cerebral infarction and edema of rats suffered from transient middle cerebral artery occlusion (tMCAO), superior to RS- and R-FMPB, as well as better than RS-FMPB by oral administration in previous studies. Importantly, S-FMPB is more active not only than the equimolar S-APB and 4-F-Eda alone or in combination but also than the clinical drugs NBP and edaravone (Eda) in combination at the equimolar doses. Furthermore, S-FMPB showed relative stability in plasma or liver microsome of rats but could be converted into two active metabolites (S-NBP and 4-F-Eda) in rats with good pharmacokinetic properties in terms of longer half-life period (t

Topics: Animals; Benzofurans; Edaravone; Ischemic Stroke; Neuroprotective Agents; Rats; Stroke

To investigate the effect of tetramethylpyrazine hydrochloride combined with butylphthalide on serum S100B, CRP, Hcy and NIHSS score in patients with acute cerebral infarction. 80 patients with acute cerebral infarction treated in our hospital from February 2019 to February 2021 were selected for retrospective analysis, and according to different treatment methods, the patients were equally divided into control group (conventional treatment) and experimental group (tetramethylpyrazine hydrochloride and butylphthalide). After treatment, the total effective rate of patients in the experimental group was significantly higher than that in the control group (P<0.05); the levels of serum S100B, CRP and Hcy, and NHISS scores in the two groups decreased, and the experimental group was significantly lower than the control group (P<0.05); the ADL scores of the two groups increased and the experimental group witnessed higher score (P<0.05); the number of patients in the experimental group with scores of 0-2 and 5 were significantly larger than that in the control group (P<0.05). The combination of tetramethylpyrazine hydrochloride and butylphthalide emanates a promising result in the treatment of patients with ACI. It reduces serum S100B, CRP and Hcy levels, protects nerve tissue, and improves nerve function, and thus merits clinical application.

Topics: Acute Disease; Benzofurans; Brain Ischemia; Cerebral Infarction; Humans; Pyrazines; Retrospective Studies; S100 Calcium Binding Protein beta Subunit; Stroke

The butylphthalide(NBP), a colorless or light yellow viscous oily component isolated from celery seeds, has the effects of anti-inflammation, anti-oxidative stress, protecting blood-brain barrier, improving cerebral microcirculation, and promoting angiogenesis. It can protect the neurological function of patients with ischemic stroke through a variety of mechanisms, improve the symptoms of patients, and contribute to the long-term recovery of them. Therefore, independently developed in China, NBP was approved by State Food and Drug Administration for the clinical treatment of stroke patients in 2002. At the same time, owing to the complex multi-target pharmacological mechanism, NBP has been frequently used in clinical practice. As frequently verified, it has obvious effects in the treatment of other neurological diseases such as Alzheimer's disease, vascular dementia, Parkinson's disease, autoimmune diseases, depression, traumatic central nervous system injury. Moreover, it demonstrates significant pharmacological effects on non-neurological diseases such as diabetes mellitus and myocardial infarction. Therefore, this study summarizes the research progress on roles of NBP in nervous system diseases and non-nervous system diseases, and the pharmacological characteristics and mechanisms of NBP, which is expected to lay a basis for research on related targets.

Topics: Benzofurans; Humans; Nervous System Diseases; Neuroprotective Agents; Oxidative Stress

Osteoarthritis (OA) has been reported as a progressive disease in the elderly, primarily characterized by degenerated articular cartilage. There has been no satisfactory drug for the treatment of OA. DL-3-n-butylphthalide (NBP), a small molecule compound extracted from celery seeds, may have antiapoptotic, antioxidant, and anti-inflammatory activities in numerous studies. However, the effects of NBP on OA and its mechanisms have been rarely reported. In this study, the effect of NBP on OA in vitro and in vivo and its possible mechanism were investigated. The results showed that NBP injection into the knee joint inhibited osteoarthritis development in a rat model of osteoarthritis induced by DMM+ACLT. NBP could increase the expressions of extracellular matrix-related components (such as type II collagen, aggrecan, proteoglycan 4, and SRY-box 9) in human osteoarthritic chondrocytes and cartilage explants. Moreover, NBP promoted the expressions of SOD and CAT. NBP upregulated the expression of FoxO3a by inhibiting the PI3K/AKT pathway, which subsequently inhibited the apoptosis of human OA chondrocytes. In conclusion, NBP promotes cartilage extracellular matrix synthesis and inhibits osteoarthritis development and the underlying mechanism related to the activation of FoxO3a.

Topics: Aged; Animals; Benzofurans; Cartilage, Articular; Chondrocytes; Extracellular Matrix; Forkhead Box Protein O3; Humans; Osteoarthritis; Phosphatidylinositol 3-Kinases; Rats

The aim of this study is to observe the effect of butylphthalide (NBP) on the amino acid content in the brain of epileptic mice.. NBP was injected intraperitoneally into the mice, and acute epileptic mice models were made after 30 minutes. The change of the four amino (aspartic acid, gamma-aminobutyric acid, glutamate, glycine) content in the brain of the epileptic mice was investigated.. The contents of Glu and Glu/GABA (control group: 38.78, NBP high-dose group: 5.52) in the NBP high-dose group were lower compared with the control group. The difference was statistically significant.. NBP could regulate the balance of excitement and inhibition systems by reducing the contents of Glu and Glu/GABA, which might relieve seizures.

Topics: Amino Acids; Animals; Benzofurans; Brain; Epilepsy; gamma-Aminobutyric Acid; Glutamic Acid; Mice

Topics: Administration, Oral; Animals; Benzofurans; Carbon Radioisotopes; Chromatography, High Pressure Liquid; Feces; Female; Male; Mass Spectrometry; Rats; Rats, Sprague-Dawley; Tissue Distribution

Cerebral infarction (CI) is the mayor reason of death in China. Reperfusion is the only immediate treatment for acute cerebral infarction. However, blood reperfusion recovery may cause ischemia-reperfusion (I/R) injuries. The purpose of this study was to investigate the effects of Tetrandrine (TTD) and 3-n-Butylphthalide (NBP) on cerebral I/R injury.. I/R was used to establish CI model

Topics: Activating Transcription Factor 2; Animals; Benzofurans; Benzylisoquinolines; Brain Ischemia; Cells, Cultured; Drug Therapy, Combination; Male; Mice; Mice, Inbred C57BL; Neuroprotective Agents; Reperfusion Injury; Signal Transduction; Toll-Like Receptor 4

DL‑3‑n‑butylphthalide (NBP) and 3‑methyl‑1- phenyl‑2‑pyrazolin‑5‑one (edaravone) are acknowledged neuroprotective agents that protect against ischemic stroke. However, the underlying mechanisms of a combination therapy with NBP and edaravone have not yet been fully clarified. The aim of the present study was to explore whether the co‑administration of NBP and edaravone had multi‑target protective effects on the neurovascular unit (NVU) of mice affected by ischemic stroke. Male C57BL/6 mice were randomly divided into the following three groups: i) Sham operation control, ii) middle cerebral artery occlusion (MCAO) and reperfusion, iii) and MCAO/reperfusion with the co‑administration of NBP (40 mg/kg) and edaravone (6 mg/kg) delivered via intraperitoneal injection at 0 and 4 h after reperfusion (NBP + edaravone). After ischemia and reperfusion, infarct volumes and neurological deficits were evaluated. The immunoreactivity of the NVU, comprising neurons, endothelial cells and astrocytes, was determined using immunofluorescence staining of neuronal nuclei (NeuN), platelet and endothelial cell adhesion molecule 1 (CD31) and glial fibrillary acidic protein (GFAP). Western blotting was used to detect the expression levels of apoptosis‑related proteins. The infarct volume, neurological function scores and cell damage were increased in the MCAO group compared with the sham operation group. Furthermore, the MCAO mice had reduced NeuN and CD31 expression and increased GFAP expression compared with the sham group. By contrast, the NBP + edaravone group exhibited reduced cell damage and consequently lower infarct volume and neurological deficit scores compared with the MCAO group. The NBP + edaravone group exhibited increased NeuN and CD31 expression and decreased GFAP expression compared with the MCAO group. Furthermore, the expression levels of Bax and cleaved caspase‑3 in the NBP + edaravone group were decreased significantly compared with the MCAO group, while the expression levels of Bcl‑2 and mitochondrial cytochrome c were increased. In conclusion, the results of the present study demonstrated that NBP and edaravone effectively prevented ischemic stroke damage with multi‑target protective effects. In addition, NBP + edaravone may be a promising combination therapy for ischemic stroke.

Topics: Animals; Apoptosis Regulatory Proteins; Benzofurans; Brain Ischemia; Disease Models, Animal; Drug Therapy, Combination; Edaravone; Endothelial Cells; Glial Fibrillary Acidic Protein; Infarction, Middle Cerebral Artery; Ischemic Stroke; Male; Mice; Mice, Inbred C57BL; Neurons; Neuroprotective Agents

3-n-Butylphthalide (NBP) is widely used for the treatment of cerebral ischaemic stroke but can causeliver injury in clinical practice. This study aims to elucidate the underlying mechanisms and propose potential preventive strategies.. NBP and its four major metabolites, 3-hydroxy-NBP (3-OH-NBP), 10-hydroxy-NBP, 10-keto-NBP and NBP-11-oic acid, were synthesized and evaluated in primary human or rat hepatocytes (PHHs, PRHs). NBP-related substances or amino acid adducts were identified and semi-quantitated by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). The target proteins and binding sites were identified by shotgun proteomics based on peptide mass fingerprinting coupled with tandem mass spectrometry and verified by molecular docking.. The toxicity of NBP and its four major metabolites were compared in both PHHs and PRHs, and 3-OH-NBP was found to be the most toxic metabolite. 3-OH-NBP induced remarkable cell death and oxidative stresses in hepatocytes, which correlated well with the levels of glutathione and N-acetylcysteine adducts (3-GSH-NBP and 3-NAC-NBP) in cell supernatants. Additionally, 3-OH-NBP covalently conjugated with intracellular Cys, Lys and Ser, with preferable binding to Cys sites at Myh9 Cys1380, Prdx4 Cys53, Vdac2 Cys48 and Vdac3 Cys36. Furthermore, we found that CYP3A4 induction by rifampicin augmented NBP-induced cell toxicity and supplementing with GSH or NAC alleviated the oxidative stresses and reactive metabolites caused by 3-OH-NBP.. Our work suggests that glutathione depletion, mitochondrial injury and covalent protein modification are the main causes of NBP-induced hepatotoxicity, which may be prevented by exogenous GSH or NAC supplementation and avoiding concomitant use of CYP3A4 inducers.

Topics: Acetylcysteine; Animals; Benzofurans; Binding Sites; Cells, Cultured; Cytochrome P-450 CYP3A Inducers; Dose-Response Relationship, Drug; Glutathione; Hepatocytes; Humans; Protein Structure, Tertiary; Rats; Rats, Sprague-Dawley

As a multi-target drug to treat ischemic stroke, N-butylphthalide (NBP) is extremely water-insoluble and exhibits limited oral bioavailability, impeding its wide oral application. Effective treatment of ischemic stroke by NBP requires timely and efficient drug exposure, necessitating the development of new oral formulations. Herein, liposomes containing biosurfactant sodium cholate (CA-liposomes) were systemically investigated as an oral NBP delivery platform because of its high biocompatibility and great potential for clinical applications. The optimized liposomes have a uniform hydrodynamic size of 104.30 ± 1.60 nm and excellent encapsulation efficiency (93.91 ± 1.10%). Intriguingly, NBP-loaded CA-liposomes produced rapid drug release and the cumulative release was up to 88.09 ± 4.04% during 12 h while that for NBP group was only 6.79 ± 0.99%. Caco-2 cell monolayer assay demonstrated the superior cell uptake and transport efficiency of NBP-loaded CA-liposomes than free NBP, which was mediated by passive diffusion via transcellular and paracellular routes. After oral administration to rats, NBP-loaded CA-liposomes exhibited rapid and almost complete drug absorption, with a t

Topics: Animals; Area Under Curve; Benzofurans; Caco-2 Cells; Chemistry, Pharmaceutical; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Carriers; Drug Liberation; Humans; Ischemic Stroke; Liposomes; Male; Neuroprotective Agents; Particle Size; Random Allocation; Rats; Rats, Sprague-Dawley; Sodium Cholate; Tissue Distribution

Neuroinflammation and mitochondrial impairment play important roles in the neuropathogenesis of Parkinson's disease (PD). The activation of NLRP3 inflammasome and the accumulation of α-synuclein (α-Syn) are strictly correlated to neuroinflammation. Therefore, the regulation of NLRP3 inflammasome activation and α-Syn aggregation might have therapeutic potential. It has been indicated that Dl-3-n-butylphthalide (NBP) produces neuroprotection against some neurological diseases such as ischemic stroke. We here intended to explore whether NBP suppressed NLRP3 inflammasome activation and reduced α-Syn aggregation, thus protecting dopaminergic neurons against neuroinflammation.. In our study, we established a MPTP-induced mouse model and 6-OHDA-induced SH-SY5Y cell model to examine the neuroprotective actions of NBP. We then performed behavioral tests to examine motor dysfunction in MPTP-exposed mice after NBP treatment. Western blotting, immunofluorescence staining, flow cytometry and RT-qPCR were conducted to investigate the expression of NLRP3 inflammasomes, neuroinflammatory cytokines, PARP1, p-α-Syn, and markers of microgliosis and astrogliosis.. The results showed that NBP exerts a neuroprotective effect on experimental PD models.. In summary, NBP rescued dopaminergic neurons by reducing NLRP3 inflammasome activation and ameliorating mitochondrial impairments and increases in p-α-Syn levels. This current study may provide novel neuroprotective mechanisms of NBP as a potential therapeutic agent.

Topics: alpha-Synuclein; Animals; Apoptosis; Benzofurans; Cell Line; Disease Models, Animal; Dopaminergic Neurons; Humans; Inflammasomes; Mice; Mitochondria; Neuroprotective Agents; NLR Family, Pyrin Domain-Containing 3 Protein; Parkinson Disease; Protein Aggregation, Pathological

Topics: Adult; Aged; Benzofurans; Cerebral Infarction; Cytokines; Drug Therapy, Combination; Female; Humans; Infusions, Intravenous; Kallikreins; Male; Middle Aged; Phosphopyruvate Hydratase; Treatment Outcome

To investigate the effects of DL-3-N-butylphthalide (NBP) via intranasal delivery after ischaemic stroke in mice.. C57BL/6 mice were divided into three groups: sham, stroke with vehicle and stroke with NBP treatment. Ischaemic stroke was induced by permanent ligation of right middle cerebral artery with 7 min common carotid artery occlusion. NBP (100 mg/kg) or vehicle was intranasally administered at 1 hour after stroke and repeated once a day until sacrifice. Bromodeoxyuridine (BrdU) (50 mg/kg/day) was given from the third day until sacrifice. Sensorimotor function was tested during 1-21 days after stroke. Local cerebral blood flow in the ischaemic and peri-infarct regions was measured using laser Doppler flowmetry before, during and 3 days after ischaemia. Expressions of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase as well as regenerative marker BrdU in the peri-infarct region were analysed by western blotting and immunohistochemical methods.. Compared with the vehicle group, NBP treatment significantly increased the VEGF expression in the poststroke brain. Stroke mice that received NBP showed significantly less vascular damage after stroke and more new neurons and blood vessels in the peri-infarct region at 21 days after stroke. In the adhesive removal test, the sensorimotor function of stroke mice treated with NBP performed significantly better at 1, 3 and 7 days after stroke compared with vehicle controls.. Daily intranasal NBP treatment provides protective and neurogenic/angiogenic effects in the poststroke brain, accompanied with functional improvements after a focal ischaemic stroke in mice.

Topics: Animals; Benzofurans; Brain Ischemia; Ischemic Stroke; Mice; Mice, Inbred C57BL; Stroke; Vascular Endothelial Growth Factor A

Alzheimer's disease (AD) is the most common cause of dementia among elderly people. Despite enormous efforts, the pathogenesis of AD still remains unclear and no drug has yet been proved to be disease-modifying. As the basis of learning and memory, the plasticity of synapse and dendritic spine has been impaired during AD progression. Previous studies have showed a protective effect of L-3-n-butylphthalide (L-NBP) on cognitive deficits in AD, we wonder whether this protective effect is associated with positive alterations on synapse and dendritic spines. In this study, we first of all confirmed the anti-dementia effect of L-NBP in 13-month-old APP/PS1 mice, and then investigated the alterations in synaptic and dendritic spine plasticity due to L-NBP treatment both in vivo and in vitro. We also conducted preliminary studies and found the possible mechanisms related to the inhibition of over-activated complement cascade and the remodeling of actin cytoskeleton. Besides, we also found extra benefits of L-NBP on presynaptic dystrophic neurites and attempted to give explanations from the view of autophagy regulation. Taken together, our study added some new evidence to the application of L-NBP in AD treatment and provided deeper insight into the relevant mechanisms for future study.

Topics: Alzheimer Disease; Animals; Autophagosomes; Autophagy; Axons; Benzofurans; Biomarkers; Cells, Cultured; Cognitive Dysfunction; Dendritic Spines; Disease Models, Animal; Hippocampus; Long-Term Potentiation; Lysosomes; Male; Mice, Transgenic; Morris Water Maze Test; Neurites; Neuronal Plasticity; Synapses

Cerebral ischemia/reperfusion (I/R) injury may lead to a poor prognosis for ischemic stroke patients after reperfusion therapy, and currently, lacks effective therapeutic intervention. This study aimed to investigate the effects of L-3-n-butylphthalide (L-NBP) on cerebral I/R injury in rats. Rat models of cerebral I/R injury were established using the middle cerebral artery occlusion/refusion (MACO/R) surgery and were administrated intragastrically with L-NBP or vehicle. We found that L-NBP attenuated the histological damages and reduced the brain hematoma in MACO/R rats. L-NBP also significantly improved the neurological function, alleviated the brain edema, and reduced the permeability of blood-brain barrier of MACO/R rats. Moreover, we detected that L-NBP considerably facilitated microvessel formation in the lesion area of brain in MACO/R rats. Finally, we found that L-NBP significantly increased the protein and mRNA expression levels of Nrf2, HIF-1α, and VEGF in the brain of MACO/R rats. In conclusion, our results demonstrated that L-NBP exerted significant beneficial effects on cerebral I/R injury in rats through promoting angiogenesis, which may be associated with the activation of Nrf2/HIF-1α/VEGF signaling pathway. Our results suggested that L-NBP could be a potential therapeutic drug for cerebral I/R injury.

Topics: Animals; Benzofurans; Brain Ischemia; Humans; Infarction, Middle Cerebral Artery; Neuroprotective Agents; Rats; Reperfusion Injury

Amyloid-β (Aβ) deposit in the parenchyma is a major characteristic in Alzheimer's disease (AD), and the impaired glymphatic clearance contributes to the Aβ accumulation. It was reported that L-3-n-butylphthalide (NBP) showed the potential neuroprotective effect in the rodent models of AD. The effects of NBP on the glymphatic system were explored in this study. In the wild-type mice, both CSF tracer influx and perivascular drainage increased after NBP treatment compared with vehicle treatment. Moreover, NBP promoted the perivascular drainage of Aβ via increased cerebral pulsation, which could be inhibited by propranolol. Then, we studied the potential of 3-month NBP treatment on Aβ deposits in 8-month-old APP/PS1 transgenic mice. NBP daily treatments remarkably improved cognitive behavior in Morris water maze. Furthermore, NBP could reduce Aβ deposition and decrease parenchymal Aβ levels. In addition, NBP markedly improved the perivascular AQP4 localization. Our results indicated that NBP could enhance the glymphatic clearance and reduce parenchymal Aβ deposit in the APP/PS1 mice, suggesting that it may have potential in the treatment of AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Benzofurans; Brain; Glymphatic System; Male; Maze Learning; Mice; Mice, Inbred C57BL; Neuroprotective Agents

Neurite outgrowth is the basis for wiring during the development of the nervous system. Dl-3-n-butylphthalide (NBP) has been recognized as a promising treatment to improve behavioral, neurological and cognitive outcomes in ischemic stroke. However, little is known about the effect and mechanism of NBP on the neurite outgrowth. In this study, we used different methods to investigate the potential effects of NBP on the neurite extension and plasticity of immature and mature primary cortical neurons and explored the underlying mechanisms. Our results demonstrated that in immature and mature cortical neurons, NBP promoted the neurite length and intersections, increased neuritic arborization, elevated numbers of neurite branch and terminal points and improved neurite complexity and plasticity of neuronal development processes. Besides, our data revealed that NBP promoted neurite extension and branching partly by activating Shh signaling pathway via increasing Gap43 expression both in immature and mature primary cortical neurons. The present study provided new insights into the contribution of NBP in neuronal plasticity and unveiled a novel pathway to induce Gap43 expression in primary cortical neurons.

Topics: Animals; Benzofurans; Cell Survival; Cells, Cultured; Female; GAP-43 Protein; Hedgehog Proteins; Mice; Mice, Inbred C57BL; Neuronal Outgrowth; Neuronal Plasticity; Neurons; Pregnancy; Signal Transduction; Up-Regulation

Dl-3-n-butylphthalide (NBP) has been widely used to treat ischemic stroke in China. To investigate the mechanisms underlying NBP activity, we established a permanent middle cerebral artery occlusion (pMCAO) rat model and injected the rats with 4 mg/kg/d NBP for nine days. We then assessed neuroinflammation, neovascularization and nerve regeneration within the brain. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI) was used to determine the phospholipid distribution, while laser ablation-inductively coupled plasma mass spectrometry imaging (LA-ICP MSI) was used to measure Foxp3, Ki-67 and pCREB levels in the brain. Immunohistochemistry was used to investigate the expression of NLR family pyrin domain containing 3 (NLRP3) and its inflammatory products, caspase-1 and interleukin-1β, in brain tissues. NBP attenuated ischemic damage and ameliorated neurological deficits in rats with pMCAO. In the ischemic brain region, NBP reduced phosphatidylethanolamine (18:0), NLRP3, caspase-1 and interleukin-1β levels, but increased levels of Foxp3, Ki-67, pCREB and several phospholipids. In molecular docking analyses, NBP bound to NLRP3, interleukin-1β, caspase-1, Foxp3, and Ki-67. These results demonstrate that NBP reduces neuroinflammation in brain tissues and promotes nerve and blood vessel regeneration, thus protecting neuromorphology and function.

Topics: Animals; Benzofurans; Brain; Caspase 1; Cyclic AMP Response Element-Binding Protein; Forkhead Transcription Factors; Infarction, Middle Cerebral Artery; Inflammation; Ischemic Stroke; Ki-67 Antigen; Molecular Docking Simulation; Neovascularization, Physiologic; Nerve Regeneration; Neuroprotective Agents; NLR Family, Pyrin Domain-Containing 3 Protein; Phospholipids; Rats; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Topics: Animals; Apoptosis; Benzofurans; Brain-Derived Neurotrophic Factor; Cognitive Dysfunction; Dendrites; Diabetes Mellitus, Type 2; Hippocampus; Male; Mice, Inbred C57BL; Morris Water Maze Test; Neuroprotective Agents; Oxidative Stress; PC12 Cells; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Signal Transduction; Synapses

Vascular factors and mitochondria dysfunction contribute to the pathogenesis of Alzheimer's disease (AD). DL-3-n-butylphthalide (NBP) has an effect in protecting mitochondria and improving microcirculation.. The aim was to investigate the effect of donepezil combined NBP therapy in patients with mild-moderate AD.. It was a prospective cohort study. 92 mild-moderate AD patients were classified into the donepezil alone group (n = 43) or the donepezil combined NBP group (n = 49) for 48 weeks. All patients were evaluated with Alzheimer's Disease Assessment Scale-Cognitive subscale (ADAS-cog), Clinician's Interview-Based Impression of Change plus caregiver input (CIBIC-plus), Alzheimer's Disease Cooperative Study-Activities of Daily Living (ADCS-ADL), and Neuropsychiatric Inventory (NPI) every 12 weeks. All patients were monitored for adverse events (AEs). The efficacy was analyzed using multivariate logistic regression analysis.. The multivariate logistic regression analysis showed that the changes of ADAS-cog score (OR = 2.778, 95% CI: [1.087, 7. 100], p = 0.033) and ADCS-ADL score (OR = 2.733, 95% CI: [1.002, 7.459], p = 0.049) had significant difference between donepezil alone group and donepezil combined NBP group, while the changes of NPI (OR = 1.145, 95% CI: [0.463, 2.829], p = 0.769), MMSE (OR = 1.563, 95% CI: [0.615, 3.971], p = 0.348) and CIBIC-plus (OR = 2.593, 95% CI: [0.696, 9.685], p = 0.156) had no significant difference. The occurrence of AEs was similar in the two groups.. Over the 48-week treatment period, donepezil combined NBP group had slower cognitive decline and better activities of daily living in patients with mild to moderate AD. These indicated that the multi-target therapeutic effect of NBP may be a new choice for AD treatment.

Topics: Activities of Daily Living; Aged; Aged, 80 and over; Alzheimer Disease; Benzofurans; Cognitive Dysfunction; Cohort Studies; Donepezil; Drug Therapy, Combination; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Neuropsychological Tests; Prospective Studies; Treatment Outcome

Studies have revealed the protective effect of DL-3-n-butylphthalide (NBP) against diseases associated with ischemic hypoxia. However, the role of NBP in animals with hypobaric hypoxia has not been elucidated. This study investigated the effects of NBP on rodents with acute and chronic hypobaric hypoxia.. Sprague-Dwaley rats and Kunming mice administered with NBP (0, 60, 120, and 240 mg/kg for rats and 0, 90, 180, and 360 mg/kg for mice) were placed in a hypobaric hypoxia chamber at 10,000 m and the survival percentages at 30 min were determined. Then, the time and distance to exhaustion of drug-treated rodents were evaluated during treadmill running and motor-driven wheel-track treadmill experiments, conducted at 5800 m for 3 days or 20 days, to evaluate changes in physical functions. The frequency of active escapes and duration of active escapes were also determined for rats in a shuttle-box experiment, conducted at 5800 m for 6 days or 27 days, to evaluate changes in learning and memory function. ATP levels were measured in the gastrocnemius muscle and malonaldehyde (MDA), superoxide dismutase (SOD), hydrogen peroxide (H. Survival analysis at 10,000 m indicated NBP could improve hypoxia tolerance ability. The time and distance to exhaustion for mice (NBP, 90 mg/kg) and time to exhaustion for rats (NBP, 120 and 240 mg/kg) significantly increased under conditions of acute hypoxia compared with control group. NBP treatment also significantly increased the time to exhaustion for rats when exposed to chronic hypoxia. Moreover, 240 mg/kg NBP significantly increased the frequency of active escapes under conditions of acute hypoxia. Furthermore, the levels of MDA and H. NBP improved physical and learning and memory functions in rodents exposed to acute or chronic hypobaric hypoxia by increasing their anti-oxidative capacity and energy supply.

Topics: Altitude Sickness; Animals; Benzofurans; Disease Models, Animal; Memory, Short-Term; Rats; Rats, Sprague-Dawley; Survival Analysis

Topics: Animals; Apoptosis; Benzofurans; Brain Ischemia; Cell Line; Cell Line, Tumor; Drug Design; Edaravone; Glucose; Humans; Hydrogen Peroxide; Hypoxia; Infarction, Middle Cerebral Artery; Male; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidative Stress; Oxygen; Platelet Aggregation; Rats; Rats, Sprague-Dawley; Reperfusion Injury

Renal ischemia reperfusion injury (IRI) has become a growing concern in clinical practice with high morbidity and mortality rates. There is currently no effective prophylactic regimen available to prevent its occurrence and to improve its clinical prognosis. Dl-3-n-butylphthalide (NBP) has been used for stroke treatment in China for years. Little is known about its role in preventing kidney injury.. The kidneys of male C57BL/6J mice were subjected to 33 min of ischemia followed by 24 h of reperfusion. NBP was administered by gavage prior to surgery. The reno-protective effect of NBP was evaluated by serum creatinine, kidney injury markers and renal pathological changes. Furthermore, the inflammation, oxidative stress, and apoptosis markers in kidney tissue were examined. In vitro, HK2 cells were treated prophylactically with NBP and then exposed to hypoxia/reoxygenation (H/R). Cell viability and apoptosis related protein were quantified to verify the protective effect of NBP. Pro-inflammation genes expression as well as ROS generation were further investigated also.. NBP pretreatment significantly improved renal dysfunction and alleviated pathological injury, renal inflammation response, oxidative stress and cell apoptosis. Consistently, NBP attenuated H/R induced increases in ROS, pro-inflammatory genes expression, apoptosis and cleaved caspase-3 levels in HK2 cells.. Our promising results validated for the first time that NBP could ameliorate renal IRI via attenuating inflammation, oxidative stress, and apoptosis, which indicated that NBP might be a good candidate against AKI.

Topics: Animals; Apoptosis; Benzofurans; Caspase 3; Cell Hypoxia; Cell Survival; Creatinine; Disease Models, Animal; Immunohistochemistry; Inflammation; Kidney; Male; Mice; Mice, Inbred C57BL; Neuroprotective Agents; Oxidative Stress; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Reperfusion Injury

As the potential risk of radiation exposure is increasing, radioprotectors studies are gaining importance. In this study, novel hybrid compounds containing edaravone analogue and 3-n-butylphthalide ring-opening derivatives were synthesized, and their radioprotective effects were evaluated. Among these, compound 10a displayed the highest radioprotective activity in IEC-6 and HFL-1 cells. Its oral administration increased the survival rates of irradiated mice and alleviated total body irradiation (TBI)-induced hematopoietic damage by mitigating myelosuppression and improving hematopoietic stem/progenitor cell frequencies. Furthermore, 10a treatment prevented abdominal irradiation (ABI)-induced structural damage to the small intestine. Experiment results demonstrated that 10a increased the number of Lgr5

Topics: Animals; Apoptosis; Benzofurans; DNA Damage; Edaravone; Epithelial Cells; Intestine, Small; Male; Mice; Mice, Inbred C57BL; NF-E2-Related Factor 2; Oxidative Stress; Radiation-Protective Agents; Whole-Body Irradiation

Hydroxypropyl-β-cyclodextrin, which possesses a high water solubility and low hemolycity, is widely used as a solubilizer and an excipient. It had also been reported that hydroxypropyl-β-cyclodextrin has the activity of regulating lipid homeostasis. In order to further understand the metabolism, the primary focus was to establish a quantitative method for hydroxypropyl-β-cyclodextrin. The analytes were extracted from plasma by protein precipitation with methanol and then carried out on a Waters CORTECS T3 column in the gradient elution of pure water and methanol. Finally, liquid chromatography-tandem mass spectrometry was applied in multiple reaction monitoring mode to complete the quantitative analysis of hydroxypropyl-β-cyclodextrin. This validated method had been successfully applied to investigate the interaction between hydroxypropyl-β-cyclodextrin and butylphthalide in vivo by optimizing the extraction reagent, simplifying the experimental procedure, and improving the sensitivity while considering the difference of drug chemical properties. Results showed that the inclusion of hydroxypropyl-β-cyclodextrin with butylphthalide significantly improved the pharmacokinetic behavior of free body hydroxypropyl-β-cyclodextrin and 3-n-butylphthalide in vivo. It had been implied that the metabolism of hydroxypropyl-β-cyclodextrin and the drug active ingredients could impact each other. It will help better application of hydroxypropyl-β-cyclodextrin and the developed method might lay the foundation for development of hydroxypropyl-β-cyclodextrin as a treatment drug for brain diseases.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Adjuvants, Pharmaceutic; Animals; Benzofurans; beta-Cyclodextrins; Chromatography, Liquid; Plasma; Platelet Aggregation Inhibitors; Rats; Tandem Mass Spectrometry

Topics: Animals; Benzofurans; Chromatography, High Pressure Liquid; Limit of Detection; Linear Models; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Tandem Mass Spectrometry

Most patients that resuscitate successfully from cardiac arrest (CA) suffer from poor neurological prognosis. DL-3-n-butylphthalide (NBP) is known to have neuroprotective effects via multiple mechanisms. This study aimed to investigate whether NBP can decrease neurological impairment after CA. We studied the protective role of NBP in the hippocampus of a rat model of cardiac arrest induced by asphyxia. Thirty-nine rats were divided randomly into sham, control, and NBP groups. Rats in control and NBP groups underwent cardiopulmonary resuscitation (CPR) 6 min after asphyxia. NBP or vehicle (saline) was administered intravenously 10 min after the return of spontaneous circulation (ROSC). Ultrastructure of hippocampal neurons was observed under transmission electron microscope. NBP treatment improved neurological function up to 72 h after CA. The ultrastructural lesion in mitochondria recovered in the NBP-treated CA model. In conclusion, our study demonstrated multiple therapeutic benefits of NBP after CA.

Topics: Animals; Apoptosis; Asphyxia; Benzofurans; Brain Diseases; Cardiopulmonary Resuscitation; Disease Models, Animal; Heart Arrest; Hippocampus; JNK Mitogen-Activated Protein Kinases; Neurons; Neuroprotection; Neuroprotective Agents; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Rats, Sprague-Dawley; Return of Spontaneous Circulation; Signal Transduction; tau Proteins; Time Factors

As a serious trauma of the neurological system, spinal cord injury (SCI) results in permanent disability, gives rise to immediate vascular damage and a wide range of matters that induce the breakage of blood spinal cord barrier (BSCB). SCI activates the expression of MMP-2/9, which are considered to accelerate the disruption of BSCB. Recent research shows that Dl-3-n-butylphthalide (NBP) exerted protective effects on blood spinal cord barrier in animals after SCI, but the underlying molecular mechanism of NBP on the BSCB undergoing SCI is unknown. Here, our research show that NBP inhibited the expression of MMP-2/9, then improved the permeability of BSCB following SCI. After the T9 level of spinal cord performed with a moderate injury, NBP was managed by intragastric administration and further performed once a day. NBP remarkably improved the permeability of BSCB and junction proteins degration, then promoted locomotion recovery. The protective effect of NBP on BSCB destruction is related to the regulation of MMP-2/9 induced by SCI. Moreover, NBP obviously inhibited the MMP-2/9 expression and junction proteins degradation in microvascular endothelial cells. In conclusion, our results indicate that MMP-2/9 are relevant to the breakdown of BSCB, NBP impairs BSCB destruction through inhibiting MMP-2/9 and promotes functional recovery subjected to SCI. NBP is likely to become a new nominee as a therapeutic to treat SCI via a transigent BSCB.

Topics: Animals; Benzofurans; Cell Hypoxia; Claudin-5; Female; Glucose; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Mice; Neuroprotective Agents; Occludin; Oxygen; Rats, Sprague-Dawley; Recovery of Function; Spinal Cord; Spinal Cord Injuries; Tight Junction Proteins; Tight Junctions

Currently, ischemic stroke is the leading cause of disability and death worldwide, and the performance of corresponding drugs is often unsatisfactory owing to the complex pathological processes and the impediment of the blood-brain barrier (BBB). Here, we employed various tertiary amino groups, including different linear, cyclic, and bimolecular drug structures, to modify 3-n-butylphthalide (NBP), a natural product used for ischemic stroke treatment, which has poor bioavailability, to generate a series of six prodrugs. These prodrugs showed significantly improved solubility and cellular uptake, which were primarily driven by putative pyrilamine cationic transporters. They also displayed more efficient brain delivery in vivo, reaching as high as 21.5-fold brain accumulation increase compared with NBP, leading to much higher bioavailability and stronger therapeutic effects. The toxicity of these molecules is also lower or similar to that of unmodified NBP. We showed that the tertiary amino group-modified NBP prodrugs are effective and safe for treating ischemic stroke with significantly enhanced druggability; hence, they have potential for further clinical development.

Topics: Benzofurans; Brain; Brain Ischemia; Humans; Ischemic Stroke; Neuroprotective Agents; Prodrugs; Stroke

Intracerebral hemorrhage (ICH) is a devastating type of stroke with high morbidity and mortality, and the effective therapies for ICH remain to be explored. L-3-n-butylphthalide (NBP) is widely used in the treatment of ischemic stroke. However, few studies evaluated the therapeutic effects of NBP on ICH. Therefore, the present study aims to evaluate the effects of NBP on ICH and its potential mechanism. The rats were randomly divided into sham-operated group, saline-treated (ICH + saline) group, and NBP-treated (ICH + NBP) group. The ICH model of SD rats induced by IV collagenase was established. The modified Garcia JH score was used to detect the neurological deficit in rats. Western Blot and immunohistochemistry analysis was applied to test the levels of UBIAD1 and caspase-3 expressions in the perihematomal region. The rates of apoptotic cells were detected by TUNEL staining. The results showed that NBP up-regulated the expression of UBIAD1, reduced the apoptotic cells in the perihematomal region, and improved the neurological deficit. Taken together, our study added some new evidence to the application of NBP in ICH treatment.

Topics: Animals; Benzofurans; Cerebral Hemorrhage; Disease Models, Animal; Hemorrhagic Stroke; Humans; Male; Neuroprotective Agents; Rats; Rats, Sprague-Dawley

Obesity is one of the world's largest health problems, and 3-N-butylphthalide (NBP), a bioactive compound in celery, has been used in dieting and weight management programs. In this study, NBP prevented high-fat-diet-induced weight gain, reduced the food efficiency ratio, altered the blood biochemical profile, and reduced the obesity-related index. NBP reduced adiposity, white fat depots, liver weight, and hepatic steatosis in obese mice. NBP ameliorated the diabetic state by decreasing glucose levels and improving glucose and insulin tolerance. NBP increased uncoupling protein-1 expression in white adipose tissue and upregulated thermogenesis by enhancing mitochondrial respiration. NBP inhibited white adipocyte development by prohibiting lipid accumulation in human adipose-derived stem cells. NBP increased free fatty acid uptake and the oxygen consumption rate in beige adipocytes. Our results suggest that NBP could be used as functional natural supplement against obesity and its associated disorders.

Topics: Adipocytes; Adipose Tissue, White; Adiposity; Animals; Benzofurans; Blood Glucose; Cells, Cultured; Diet, High-Fat; Fatty Liver; Humans; Lipid Metabolism; Lipids; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Obesity; Protective Agents; Thermogenesis

To investigate the neuroprotective effect and mechanism of DL-3-n-butylphthalide (NBP) on the brain-derived neurotrophic factor (BDNF)/tyrosine kinase B (TrkB) and its downstream signalling pathway after cerebral ischemia/reperfusion injury (CIRI) in rats.. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was used. Reperfusion was performed 2 h after ischemia, and 20 mg/kg of NBP was intraperitoneally injected. Neurological defect score and pathological changes were performed. Apoptotic cells were detected using in situ end-labelling with TUNEL. The expression of BDNF and TrkB proteins was measured by Western blot and immunohistochemical staining. BDNF mRNA, TrkB mRNA, protein kinase B (AKT) mRNA and caspase-3 mRNA expression were measured using real-time polymerase chain reaction (qPCR).. After 24 h of reperfusion, the neurological defect score and the percentage of apoptotic cells in the ischemia/reperfusion group (I/R group) were higher than those in the ischemia/reperfusion + drug group (I/R + d group). The positive expressions of BDNF and TrkB mRNA and protein in the I/R + d group were obviously higher than those in the I/R group (p < 0.05). After intervention with the TrkB receptor inhibitor (K252a), the expression levels of BDNF and TrkB and AKT mRNA were significantly decreased in the ischemia/reperfusion + drug + TrkB receptor inhibitor group (I/R + d + R group) compared with the I/R + d group, however the caspase-3 mRNA expression level showed the reverse trend. The expressions of BDNF, TrkB and p-Akt proteins in the I/R + d group were remarkably higher than those in the I/R group at each time point, and reached the peak at 24 hours after reperfusion, which were earlier than that in the I/R group.. Butylphthalide represents a neuroprotective effect after CIRI in rats and used within 24 h of early onset contributes to better prognosis. The underlying mechanism may be related to reducing the apoptosis of nerve cells through BDNF/TrkB signalling pathway.

Topics: Animals; Benzofurans; Brain Ischemia; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Reperfusion Injury

Topics: Benzofurans; Heart Arrest; Humans; Hypoxia-Ischemia, Brain

This paper aimed to discuss the denoising ability of magnetic resonance imaging (MRI) images based on fuzzy C-means clustering (FCM) algorithm and the influence of Butylphthalide combined with Edaravone treatment on nerve function and vascular endothelial function in patients with acute cerebral infarction (ACI). Based on FCM algorithm, Markov Random Field (MRF) model algorithm was introduced to obtain a novel algorithm (NFCM), which was compared with FCM and MRF algorithm in terms of misclassification rate (MCR) and difference of Kappa index (KI). 90 patients with ACI diagnosed in hospital from December 2018 to December 2019 were selected as subjects, who were divided into combined treatment group (conventional treatment + Edaravone + Butylphthalide) and Edaravone group (conventional treatment + Edaravone) randomly, each consisting of 45 cases. The National Institutes of Health Stroke Scale (NIHSS) score and endothelial function index level such as plasma nitric oxide (NO), human endothelin-1 (ET-1), and vascular endothelial cell growth factor (VEGF) were compared before and after treatment between the two groups. The results showed that the MCR of NFCM was evidently inferior to FCM and MRF, and the KI was notably higher relative to the other two algorithms. After treatment, the NIHSS score of the combined treatment group was (9.09 ± 1.86) points and that of Edaravone group was (14.97 ± 3.44) points, with evident difference between the two groups (

Topics: Algorithms; Benzofurans; Biomarkers; Cerebral Infarction; Drug Therapy, Combination; Edaravone; Endothelium, Vascular; Female; Follow-Up Studies; Fuzzy Logic; Humans; Image Interpretation, Computer-Assisted; Magnetic Resonance Imaging; Male; Middle Aged; Neuroprotective Agents; Prognosis

3-n-Butylphthalide (NBP), an extract from seeds of Apium graveolens Linn. (Chinese celery), has been demonstrated to have antidepressant effects in suspension chronic-stressed rats by our group. The purpose of this study was to investigate the possible involvement of brain-derived neurotrophic factor (BDNF) and mammalian target of rapamycin (mTOR) in the antidepressant mechanism of NBP. Chronic unpredictable mild stress (CUMS) was applied for 6 weeks to induced a depressive-like behavior, characterized by decreased locomotor activity, sucrose preference and the NE, DA and 5-HT levels in cortex. Oral treatment with NBP (30 or 100 mg/kg, p.o.), similarly to fluoxetine (2 mg/kg, p.o.), can prevention of these alterations. The NBP (30 or 100 mg/kg, p.o.) reversed the decrease in the BDNF, p-ERK, mTOR and synapsin-1 protein levels in rat cortex caused by CUMS. And rapamycin, an mTOR inhibitor, completely inhibited the antidepressant-like activity of NBP in vivo. In conclusion, these findings indicate that NBP treatment attenuated the depression-like behaviors through the modulation of serotonergic system and BDNF-ERK-mTOR signaling in rat.

Topics: Animals; Antidepressive Agents; Benzofurans; Brain-Derived Neurotrophic Factor; Cerebral Cortex; Chronic Disease; Dose-Response Relationship, Drug; Male; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Stress, Psychological; TOR Serine-Threonine Kinases; Treatment Outcome

To compare the e!cacy and functional outcomes of dl-3-n-Butylphthalide (NBP) and human urinary kallidinogenase (HUK) on ischemic stroke patients and to determine their effects on serum tumor necrosis factor-alpha (TNF-α) and vascular endothelial growth factor (VEGF).. A prospective study was conducted on 57 ischemic stroke patients. Functional outcomes were assessed by the National Institute Health Stroke Scale (NIHSS), the modified Rankin Scale (mRS), and the activities of daily living score (ADL), whereas TNF-α and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA).. TNF-α was significantly down-regulated in the NBP group and upregulated in the control group two weeks after treatment (. Both treatments are effective and can significantly promote recovery in stroke patients. Additionally, both options have similar effects in promoting long-term recovery, with NBP exerting a greater impact on serum VEGF and TNF-α expressions.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Benzofurans; Biomarkers; Brain Ischemia; Case-Control Studies; Female; Humans; Ischemic Stroke; Kallikreins; Male; Middle Aged; Neuroprotective Agents; Prognosis; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Young Adult

Blood-brain barrier (BBB) disruption and neuronal apoptosis are important pathophysiological processes after traumatic brain injury (TBI). In clinical stroke, Dl-3n-butylphthalide (Dl-NBP) has a neuroprotective effect with anti-inflammatory, anti-oxidative, anti-apoptotic and mitochondrion-protective functions. However, the effect and molecular mechanism of Dl-NBP for TBI need to be further investigated. Here, we had used an animal model of TBI and SH-SY5Y/human brain microvascular endothelial cells to explore it. We found that Dl-NBP administration exerts a neuroprotective effect in TBI/OGD and BBB disorder, which up-regulates the expression of tight junction proteins and promotes neuronal survival via inhibiting mitochondrial apoptosis. The expressions of autophagy-related proteins, including ATG7, Beclin1 and LC3II, were significantly increased after TBI/OGD, and which were reversed by Dl-NBP treatment both in vivo and in vitro. Moreover, rapamycin treatment had abolished the effect of Dl-NBP for TBI recovery. Collectively, our current studies indicate that Dl-NBP treatment improved locomotor functional recovery after TBI by inhibiting the activation of autophagy and consequently blocking the junction protein loss and neuronal apoptosis. Dl-NBP, as an anti-inflammatory and anti-oxidative drug, may act as an effective strategy for TBI recovery.

Topics: Animals; Apoptosis; Autophagy; Benzofurans; Blood-Brain Barrier; Brain Injuries, Traumatic; Cells, Cultured; Disease Models, Animal; Male; Mice; Mice, Inbred C57BL; Neuroprotective Agents; Recovery of Function

Major depressive disorder (MDD) is a severe mental disorder associated with high rates of morbidity and mortality. Current first-line pharmacotherapies for MDD are based on enhancement of monoaminergic neurotransmission, but these antidepressants are still insufficient and produce significant side-effects. Consequently, the development of novel antidepressants and therapeutic targets is desired. Dl-3-n-butylphthalide (NBP) is a compound with proven efficacy in treating ischemic stroke, yet its therapeutic effects and mechanisms for depression remain unexplored. The aim of this study was to investigate the effect of NBP in a chronic social defeat stress model of depression and its underlying molecular mechanisms. Here, we examined depression-related behavior and performed a targeted metabolomics analysis. Real-time quantitative polymerase chain reaction and western blotting were used to examine key genes and proteins involved in energy metabolism and the AKT/cAMP response element-binding protein (CREB) signaling pathway. Our results reveal NBP attenuates stress-induced social deficits, anxiety-like behavior and despair behavior, and alters metabolite levels of glycolysis and tricarboxylic acid (TCA) cycle components. NBP affected gene expression of key enzymes of the TCA cycle, as well as protein expression of p-AKT and p-CREB. Our findings provide the first evidence showing that NBP can attenuate stress-induced behavioral deficits by modulating energy metabolism by regulating activation of the AKT/CREB signaling pathway.

Topics: Animals; Benzofurans; Cyclic AMP Response Element-Binding Protein; Depressive Disorder, Major; Energy Metabolism; Mice; Neuroprotective Agents; Proto-Oncogene Proteins c-akt; Signal Transduction; Social Defeat

Diffuse leptomeningeal glioneuronal tumor with the first symptom of acute rapid blindness in both eyes are rare in adolescents. We present a case of an adolescent female without a history of cancer who developed a dramatic loss of vision and eventually blindness, and was diagnosed as diffuse leptomeningeal glioneuronal tumor by cerebrospinal fluid cytology and enhanced MRI. In order to save vision, under the condition of ineffective dehydration treatment, timely emergency surgery to implant Ommaya sac drainage cerebrospinal fluid. According to the pathophysiological mechanism of cerebral edema, we added Edaravone to scavenge oxygen free radicals, Alprostadil to improve microcirculation, Monosialotetrahexosylganglioside nutrient nerve, Butylphthalide to promote collateral circulation, combined with hyperbaric oxygen and acupoint injection of Anisodine. Finally, the patient's vision returned to normal. Conclusion: when dehydration treatment is ineffective, timely surgery to reduce intracranial pressure, combined with comprehensive treatment, can effectively save vision.

Topics: Adolescent; Benzofurans; Blindness; Cerebrospinal Fluid Shunts; Female; Humans; Meningeal Neoplasms; Neuroprotective Agents

An experimental animal study of treatment of spinal cord injury (SCI).. This report aims to evaluate the in vivo effects of butylphthalide NBP on SCI biology and to explore its potential mechanism.. SCI causes great damage to humans. The inflammatory and reconstructive processes after SCI is regulated by activation of astroglial and microglial cells. Activated microglia/macrophages can be divided into M2 (anti-inflammatory) and M1 (pro-inflammatory) phenotypes. Butylphthalide (3-n-butylphthalide or NBP) treatment can significantly alleviate ischemic brain damage, and further study has confirmed that central neuroprotective effects can be realized by converting M1 polarized microglia/macrophages to the M2 phenotype. Thus far, it remains unknown whether NBP can modulate the transition of macrophages/microglia between the M1 and M2 phenotypes.. We randomly divided male mice into three groups (sham group, SCI group, SCI+ NBP group). Molecular and histological tests were performed to detect the macrophage/microglia polarization as well as the potential mechanism of NBP in vivo and in vitro.. It was found that NBP treatment significantly attenuated the motor dysfunction and neuronal apoptosis induced by SCI. Treatment with NBP could also reduce pro-inflammatory cytokine release after SCI and could facilitate macrophage/microglia M2 polarization and inhibit M1 polarization after SCI. To verify the findings in animal experiments, we examined the effect of NBP on BV2 cell polarization, the results showed that NBP treatment could enhance M2 polarization and inhibit M1 polarization, and that M2 polarization occurred in a p38-dependent manner.. NBP plays an important role in the anti-inflammatory response in SCI via the facilitation of macrophage/microglia M2 polarization as well as the inhibition of macrophage/microglia M1 polarization. The M2 polarization of macrophages/microglia occurs via activation of p38 pathway.. 3.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Benzofurans; Cell Polarity; Macrophages; Male; Mice; Mice, Inbred C57BL; Microglia; Neuroprotective Agents; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Spinal Cord Injuries; Thoracic Vertebrae

Carbon monoxide (CO) poisoning is a common health problem among people in many countries, primarily because of its severe clinical effects and high toxicological morbidity and mortality. Acute brain injury and delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) are the most common neurological complications. This study was performed to assess the efficacy of N-butylphthalide (NBP) and dexamethasone (DXM) combined with hyperbaric oxygen (HBO) in patients with DEACMP.. A total of 171 patients with DEACMP were recruited and assigned to the combined therapy group (receiving NBP and DXM 5 mg/day plus HBO therapy) or the control group (HBO therapy as monotherapy). Conventional treatments were provided for all patients. The cognition and movement changes in patients were evaluated by the Mini-Mental State Examination (MMSE), the Montreal Cognitive Assessment (MoCA) scale and the Barthel index of activities of daily living (ADL) before and after the treatment at 1 month, 3 months, and 1 year, respectively.. At 1 month, 3 months, and 1 year after the treatment, the MMSE, MoCA and ADL scores were all significantly higher in the combined therapy group than those in the control group. There were no significant alterations in blood glucose, blood lipids, or liver and kidney function during the whole treatment session. Some patients experienced loss of appetite, mild headache and minor skin irritations. However, these patients recovered by themselves and needed no additional medications or special treatment.. These results indicated that NBP and DXM combined with HBO for the treatment of DEACMP can significantly improve the cognitive and motor functions of patients and is very safe.

Topics: Activities of Daily Living; Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Benzofurans; Brain Diseases; Carbon Monoxide Poisoning; Combined Modality Therapy; Dexamethasone; Female; Humans; Hyperbaric Oxygenation; Male; Middle Aged; Young Adult

Neurodegenerative diseases are irreversible conditions that result in progressive degeneration and death of nerve cells. Although the underlying mechanisms may vary, oxidative stress is considered to be one of the major causes of neuronal loss. Importantly, there are still no comprehensive treatments to completely cure these diseases. Therefore, protecting neurons from oxidative damage may be the most effective therapeutic strategy. Here we report a neuroprotective effects of a novel hybrid compound (dlx-23), obtained by conjugating α-lipoic acid (ALA), a natural antioxidant agent, and 3-

Topics: Benzofurans; Cell Line, Tumor; Hydrogen Peroxide; Neurons; Neuroprotective Agents; Oxidative Stress; Oxidopamine; Reactive Oxygen Species; Thioctic Acid

Vascular cognitive impairment (VCI) is a type of cerebral vascular disorder that leads to learning and memory decline. VCI models can be induced by chronic cerebral hypoperfusion via permanent bilateral common carotid artery occlusion. 3‑N‑Butylphthalide (NBP) is a neuroprotective drug used for the treatment of ischemic cerebrovascular diseases. Silent information regulator 1 (SIRT1) plays an important role in memory formation and cognitive performance, and its abnormal reduction is associated with cognitive dysfunction in neurodegenerative diseases. Brain‑derived neurotrophic factor (BDNF) is a neurotrophic factor that plays critical roles in promoting neuronal growth and injury repair. The present study was performed to investigate the effects and the underlying mechanism of NBP on learning deficits in a rat model of VCI. Rats were divided into a control group, model group, low‑NBP‑dose group (30 mg/kg/day), high‑NBP‑dose group (60 mg/kg/day), NBP + SIRT1 inhibitor group and NBP + BDNF inhibitor group. Rats were then subjected to Morris water maze and T‑maze tests, which identified that NBP treatment significantly attenuated memory impairments in VCI rats. Molecular examination indicated that SIRT1 and BDNF expression levels in the hippocampus were increased by NBP treatment. However, NBP failed to ameliorate cognitive function after inhibition of the SIRT1/BDNF signaling pathway. In addition, NBP in combination with a SIRT1 inhibitor suppressed BDNF protein expression, but inhibition of BDNF did not inhibit SIRT1 protein expression in rats with VCI. The present results suggested that the neuroprotective effects of NBP on learning deficits in a rat model of VCI may be via regulation of the SIRT1/BDNF signaling pathway, in which SIRT1 may be the upstream signaling molecule. Therefore, the SIRT1/BDNF pathway could be a potential therapeutic target for VCI.

Topics: Animals; Benzofurans; Brain-Derived Neurotrophic Factor; Cognitive Dysfunction; Dementia, Vascular; Male; Maze Learning; Memory; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Signal Transduction; Sirtuin 1

This study will specifically investigate the effect of butylphthalide on hemodynamics in patients with watershed stroke (WS).. We will search the following databases from their inceptions to the March 1, 2020: Cochrane Library, MEDLINE, EMBASE, PsycINFO, Web of Science, Cumulative Index to Nursing and Allied Health Literature, and China National Knowledge Infrastructure. All relevant randomized controlled trials on exploring the effect of butylphthalide on hemodynamics in patients with WS will be considered for inclusion. No language limitation will be imposed to this study. All study quality will be checked using Cochrane risk of bias tool. RevMan 5.3 software will be utilized for data analysis.. This study will summarize the latest evidence to investigate the effect of butylphthalide on hemodynamics in patients with WS.. Findings from this study will provide theoretical basis of butylphthalide on hemodynamics in patients with WS for clinician and future research.. This study is carried out based on the published data, thus, no ethical approval is required. We will submit this study to a peer-reviewed journal for publication.. INPLASY 202030006.

Topics: Benzofurans; China; Hemodynamics; Humans; Meta-Analysis as Topic; Neuroprotective Agents; Quality of Life; Randomized Controlled Trials as Topic; Stroke; Systematic Review as Topic; Treatment Outcome

Elevated inflammation is commonly observed in depression, but whether this association is causal is not determined. Our previous basic research indicated that Dl-3-n-butylphthalide (NBP) possessed an anti-inflammatory effect. Additional recent evidence consistently suggests that depression is associated with lipid metabolism. Therefore, our study performed an untargeted lipidomics approach of ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS) to reveal the potential discriminating lipid profile of the hippocampus for NBP involvement in lipopolysaccharide (LPS)-induced depression. Male Sprague-Dawley(SD) rats were randomly allocated to one of three groups (n = 6): control, LPS-induced model of depression (LPS), or NBP involvement in the LPS-induced model of depression (LPS + NBP). Statistical analysis was used to identify differential hippocampus lipids in the LPS, NBP + LPS, and control groups. Our study demonstrated that most of the differentially expressed lipid metabolites were involved in glycerophospholipid metabolism, sphingolipid metabolism, glycerolipid metabolism, and glycosylphosphatidylinositol(GPI)-anchor biosynthesis, which may partially account for the pathophysiological process of depression. However, more pre-clinical and clinical evidence is warranted to determine the extent and consistency of the role of NBP and further elucidate the pathophysiological mechanisms underlying inflammation-induced depression.

Topics: Animals; Benzofurans; Depression; Hippocampus; Inflammation; Lipid Metabolism; Lipidomics; Lipopolysaccharides; Male; Neuroprotective Agents; Rats; Rats, Sprague-Dawley

Cerebral stroke occurs following ischemic and hemorrhagic lesions in the brain. Survival and recovery of stroke patients depend on the severity of the initial injury but also the therapeutic approaches applied for emergent lifesaving and continuing post-stroke management. Dl-3-n-Butylphthalide (NBP), an active compound derived from Chinese celery seeds, has shown clinical efficacy in the treatment of ischemic cerebral stroke.. In the present study we explored the therapeutic effect of NBP in a rat model of intracerebral hemorrhage (ICH), focusing on its potential role in promoting neovascularization in the perihemorrhagic zone. ICH was induced in male Sprague-Dawley rats by unilateral injection of autologous blood into the globus pallidus, with sham-operated (Sham group), vehicle-treated (ICH) and NBP-treated (at 10 and 25 mg/kg/Bid, p.o., ICH + NBP10 and ICH + NBP25, respectively) groups examined behaviorally, macroscopically, histologically and biochemically at 1, 3, 7 and 15 days (d) post operation. Rats in the ICH + NBP10 and ICH + NBP25 groups showed reduced Longa's motor scores relative to the ICH groups at the 3 and 7d time points, while the hematoma volume was comparable in the two NBP relative to the ICH groups as measured at 7d and 15d. In the perihemorrhagic zone, the numeric density of blood vessels immunolabeled by CD34, an angiogenic marker, was greater in the ICH + NBP10 and ICH + NBP25 than ICH groups, more so in the higher dosage group, at 1, 3, 7 and 15d. Levels of the vascular endothelial growth factor (VEGF) and angiopoietins-2 (Ang-2) proteins were elevated in the NBP groups relative to the sham and vehicle controls in immunoblotting of tissue lysates from the injection region.. These results suggest that NBP can alleviate neurological defects following experimentally induced local brain hemorrhage, which is associated with a potential role of this drug in promoting neovascularization surrounding the bleeding loci.

Topics: Animals; Benzofurans; Brain; Cerebral Hemorrhage; Disease Models, Animal; Male; Nervous System Diseases; Rats, Sprague-Dawley; Stroke; Vascular Endothelial Growth Factor A

Topics: Animals; Anti-Inflammatory Agents; Apium; Benzofurans; Blood-Brain Barrier; Brain Edema; Cerebral Hemorrhage; Disease Models, Animal; Humans; Infusions, Intraventricular; Male; Matrix Metalloproteinase 9; Neuroprotective Agents; Permeability; Rats; Signal Transduction; Tumor Necrosis Factor-alpha

To investigate whether dl-3-n-butylphthalide (NBP) affects cholinergic system function and ameliorates cognitive decline in a rat model of vascular dementia (VaD).. The VaD rat model was established by bilateral common carotid artery ligation (two-vessel occlusion, 2VO). Rats were divided into five groups: control, sham, 2VO, 2VO+NBP (80 mg/kg; intragastric), and 2VO+donepezil (1 mg/kg; intragastric). Treatments were administered once daily for 2 weeks from day 21 post-surgery. Spatial learning and memory were evaluated by Morris water maze performance. Hippocampal choline acetyltransferase (ChAT), acetylcholinesterase (AChE), vesicular acetylcholine transporter (VAChT), vascular endothelial growth factor (VEGF), and brain-derived neurotrophic factor (BDNF) expressions were detected using immunohistochemistry, immunofluorescence, and real-time polymerase chain reaction methods.. The daily escape latency was significantly longer in 2VO rats than in the sham or control groups, while the time spent in the target quadrant was significantly shorter. The daily escape latency of the 2VO+NBP group was significantly shorter compared with the 2VO group. Following NBP treatment, ChAT, AChE, VAChT, and BDNF expressions were significantly upregulated in the hippocampus.. Central cholinergic dysfunction may be involved in VaD pathogenesis. NBP treatment significantly improved spatial learning and memory in VaD rats, and may enhance cholinergic system function via BDNF-mediated neuroprotection.

Topics: Acetylcholinesterase; Animals; Benzofurans; Brain Ischemia; China; Choline O-Acetyltransferase; Cholinergic Agents; Cholinergic Neurons; Cognitive Dysfunction; Dementia, Vascular; Disease Models, Animal; Hippocampus; Male; Maze Learning; Memory; Oxidative Stress; Rats; Rats, Sprague-Dawley; Vascular Endothelial Growth Factor A

3-

Topics: Animals; Apium; Asthma; Benzofurans; Brain Ischemia; Humans; Neuroprotective Agents; Seeds; Stereoisomerism; Stroke

Dl-3-n-butylphthalide (NBP) has been demonstrated to exert neuroprotective effects in experimental models and human patients. This study was performed to assess the therapeutic effects and the underlying molecular mechanisms of NBP in a neonatal hypoxic-ischemic rat model. The results showed that NBP treatment significantly reduced the infarct volume, improved histological recovery, decreased neuronal cell loss, enhanced neuronal cell rehabilitation, promoted neurite growth and decreased white matter injury. In addition, NBP treatment effectively improved long-term neurobehavioral development and prognosis after HI injury. We further demonstrated an inhibitory effect of NBP on endoplasmic reticulum (ER) stress-induced apoptosis, evidenced by reduction in ER stress-related protein expressions (GRP78, XBP-1, PDI and CHOP), decrease in TUNEL-positive cells, down-regulation in pro-apoptosis protein (Bax and cleaved caspase-3), up-regulation in anti-apoptosis protein (Bcl-2). Moreover, NBP exerted a protective effect in blood-brain barrier disruption, which ameliorated brain edema and reduced the degeneration of the tight junction proteins (Occludin and Claudin-5) and adherens junction proteins (P120-Catenin, VE-Cadherin and β-Catenin). Overall, our findings demonstrated that NBP treatment attenuated HI brain injury through inhibiting ER stress-induced apoptosis and alleviating blood-brain barrier disruption in newborn rats. This work provides an effective therapeutic strategy to reduce brain damage and enhance recovery after neonatal HI brain injury.

Topics: Animals; Animals, Newborn; Apoptosis; Benzofurans; Blood-Brain Barrier; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Hypoxia-Ischemia, Brain; Neuroprotective Agents; Oxidative Stress; Rats; Rats, Sprague-Dawley; Tight Junction Proteins

To evaluate the effect of dl-3-N-butylphthalide (NBP) on new cerebral microbleeds (CMBs) in patients with acute ischemic stroke (AIS).. We will prospectively enroll patients with AIS admitted to the stroke center of Jingjiang People's Hospital. Qualified participants will be randomly assigned to either the NBP group (NBP injection) or the control group (NBP injection placebo) in a ratio of 1:1. Patients will complete the brain magnetic resonance imaging within 48 hours and 14 days after stroke onset to observe the CMBs through susceptibility weighted imaging, and evaluate whether the use of NBP will affect the new CMBs in AIS patients. SPSS 20.0 will be used for statistical analyses.. We will provide practical and targeted results assessing the safety of NBP for AIS patients, to provide reference for clinical use of NBP.. The stronger evidence about the effect of NBP on new CMBs in AIS patients will be provided for clinicians.

Topics: Benzofurans; Cerebral Hemorrhage; Clinical Protocols; Humans; Ischemia; Magnetic Resonance Imaging; Platelet Aggregation Inhibitors; Prospective Studies; Stroke

Observe the clinical efficacy of l-3-n-Butylphthalide (NBP) in acute ischemic stroke (AIS) patients within 24 h after intravenous thrombolysis using recombinant tissue plasminogen activator (hereafter termed "IT").. One-hundred and seventy-eight patients with AIS were divided randomly into two groups: NBP and control. The former was given a NBP injection within 24 h after IT. After intravenous injection of NBP for 8-10 days, patients switched to soft capsules of NBP before or during meals. NBP treatment was continued for ≥30 days after hospital discharge. In the control group, NBP was not injected within 24 h after IT, and NBP capsules were not given after 8-10 days. Both groups were reviewed for CT or MRI 24 h after IT. The National Institutes of Health Stroke Scale (NIHSS) score was calculated. The number of patients with a modified Rankin scale (mRS) 0-2 before, 24 h, and 90 days after IT was documented. Prevalence of cerebral hemorrhage and reocclusion of blood vessels after IT was calculated.. There were no differences in sex, age, blood pressure, blood glucose, or cerebral-infarction types between the two groups before treatment. The NIHSS score 24 h after IT and the percentage of mRS scores 0-2 were not significantly different between the two groups. Compared with the control group, the NIHSS score in the NBP group was significantly improved at 90 days, and the number of patients with a mRS score 0-2 increased significantly. There was no significant difference in hemorrhage prevalence after IT between the two groups. Prevalence of blood-vessel occlusion after IT was significantly lower in the NBP group than that in the control group.. Use of NBP within 24 h after IT can reduce the prevalence of reocclusion of blood vessels without increasing the risk of cerebral hemorrhage.

Topics: Administration, Intravenous; Adult; Aged; Aged, 80 and over; Benzofurans; Capsules; Cerebral Infarction; Female; Humans; Male; Middle Aged; Thrombolytic Therapy; Time Factors; Tissue Plasminogen Activator

Neuromyelitis optica spectrum disorders (NMOSDs) are inflammatory demyelinating disorders of the central nervous system; they are characterized by severe optic neuritis and transverse myelitis. Intravenous methylprednisolone pulse (IVMP) therapy is an effective treatment that is administered to patients in the acute phase of NMOSD; this therapy has achieved remarkable results in clinical practice. However, there are no reports on NMOSD patients who have experienced an acute bilateral cerebral infarction while undergoing IVMP treatment.. We report on a 62-yr-old woman who was undergoing IVMP therapy for the primary diagnosis of NMOSD. Unexpectedly, the patient's existing limb weakness worsened, and she developed motor aphasia on the second day of IVMP treatment. Additionally, brain magnetic resonance imaging revealed acute bilateral cerebral infarction.. The patient's clinical manifestations, medical imaging results, and laboratory test results were taken into consideration; the final diagnosis was acute bilateral cerebral infarction in the presence of NMOSD.. Subsequent to the onset of acute cerebral infarction, the patient was immediately treated with oral aspirin, atorvastatin, and intravenous butylphthalide. The hormone dose was adjusted to an oral 60-mg/d dose for maintenance; this was followed by immunoadsorption plasmapheresis for 3 days, and double-filtration plasmapheresis for 2 days.. Following treatment onset, the patient's ocular symptoms significantly improved, and her limb muscle strength gradually recovered. Two months after discharge, the patient's husband reported that she was able to walk with the help of others and take care of herself, and that there was no recurrence.. Medical professionals must be aware of the possibility of NMOSD patients with cerebrovascular risk factors suffering an acute cerebral infarction while undergoing high-dose IVMP therapy, as this therapy can exacerbate existing problems.

Topics: Acute Disease; Administration, Intravenous; Administration, Oral; Anticholesteremic Agents; Aphasia, Broca; Aspirin; Atorvastatin; Benzofurans; Cerebral Infarction; Female; Humans; Magnetic Resonance Imaging; Methylprednisolone; Middle Aged; Neuromyelitis Optica; Neuroprotective Agents; Plasmapheresis; Platelet Aggregation Inhibitors; Treatment Outcome

This experiment was aimed to investigate the effect of Dengzhan Shengmai capsule combined with butylphthalide soft capsule on oxidative stress indexes and serum homocysteine (Hcy) and C-reactive protein (CRP) levels in patients with vascular dementia (VD). From July 2017 to July 2019, 123 patients with VD in our hospital were selected as the research object, and each patient was assigned a random number according to the order of treatment. Among them, No. 1 to 41 were the control group A, No. 42 to 82 were the control group B, and No. 83 to 123 were the research group. Control group A was given butylphthalide soft capsules, control group B was Dengzhanshengmai capsules, and the research group was given Dengzhanshengmai capsules combined with butylphthalide softgels. Comparison of clinical efficacy, the incidence of adverse reactions, and improvement of symptoms [Montreal Cognitive Assessment Scale (MocA) score, Vascular Dementia TCM Syndrome Differentiation Scale (SDSVD) score], vascular endothelial function [NO, endothelin 1 (ET-1)], oxidative stress [lipid peroxide (LPO), superoxide dismutase (SOD), C Dialdehyde (MDA)], endoplasmic reticulum stress response (Hcy, CRP) related indicators before and after treatment in three groups. Results showed that the total effective rate of treatment in the study group was higher than that in the control groups A and B, the difference was statistically significant (p<0.05). In symptom improvement, the MoCA score of the study group was higher than that of the control groups A and B after the treatment course, and the SDSVD score was lower than that of the control groups A and B (p<0.05); In the vascular endothelial function section, after the course of treatment, the serum NO level in the study group was higher than that in the control groups A and B, and the ET-1 level was lower than that in the control groups A and B (p<0.05). In oxidative stress experiment, after the course of treatment in the study group, the serum LPO and MDA levels were lower than those in the control groups A and B, and the SOD levels were higher than those in the control groups A and B (p<0.05).  Endoplasmic reticulum stress response results showed that after the course of treatment in the study group, the serum Hcy and CRP levels were lower than those in the control groups A and B (p<0.05). In adverse reactions section, there was no significant difference in the incidence of adverse reactions among the three groups (p>0.05).

Topics: Aged; Benzofurans; C-Reactive Protein; Dementia, Vascular; Drugs, Chinese Herbal; Female; Homocysteine; Humans; Male; Malondialdehyde; Middle Aged; Oxidation-Reduction; Oxidative Stress; Reactive Oxygen Species; Superoxide Dismutase

Butylphthalide is widely used for the adjunctive treatment of vascular dementia; however, the clinical evidences are not well synthesized yet.. We proposed a systematic review and meta-analysis to evaluate the efficacy and safety of butylphthalide as adjunctive therapy for vascular dementia. Seven electronic databases (China National Knowledge Infrastructure, Wanfang database, Chongqing VIP database, China Biomedical Literature Database, Pubmed, EMBASE and Cochrane library) will be searched for eligible randomized controlled trials (RCTs). Required data of included studies will be collected. Quality of studies will be evaluated using Cochrane risk of bias assessment tool. Data synthesis will be performed using Review Manager software. Subgroup analysis and sensitivity analysis will also be carried.. Synthesis results of current available RCTs regarding the efficacy and safety of butylphthalide for the treatment vascular dementia will be provided by this systematic review and meta-analysis.. This systematic review and meta-analysis will provide high level evidence of butylphthalide clinical application.. PROSPERO CRD42020168947.

Topics: Benzofurans; Clinical Protocols; Dementia, Vascular; Humans; Meta-Analysis as Topic; Platelet Aggregation Inhibitors; Randomized Controlled Trials as Topic; Systematic Reviews as Topic; Treatment Outcome

dl-3-n-butylphthalide (dl-NBP) is a powerful antioxidant compound with profound neuroprotective effects in stroke and brain injury. However, its role in Parkinson's disease (PD) is not well known. Traumatic brain injury (TBI) is one of the key factors in precipitating PD like symptoms in civilians and particularly in military personnel. Thus, it would be interesting to explore the possible neuroprotective effects of NBP in PD following concussive head injury (CHI). In this chapter effect of nanowired delivery of NBP together with mesenchymal stem cells (MSCs) in PD with CHI is discussed based on our own investigations. It appears that CHI exacerbates PD pathophysiology in terms of p-tau, α-synuclein (ASNC) levels in the cerebrospinal fluid (CSF) and the loss of TH immunoreactivity in substantia niagra pars compacta (SNpc) and striatum (STr) along with dopamine (DA), dopamine decarboxylase (DOPAC). And homovanillic acid (HVA). Our observations are the first to show that a combination of NBP with MSCs when delivered using nanowired technology induces superior neuroprotective effects in PD brain pathology exacerbated by CHI, not reported earlier.

Topics: Benzofurans; Brain Injuries, Traumatic; Humans; Mesenchymal Stem Cells; Neuroprotection; Parkinson Disease; Titanium

Butylphthalide (NBP) is a phthalide compound contained in Angelicae Sinensis Radix which is one of the most widely used traditional Chinese medicines. This study aims to explore the therapeutic effect of NBP on airway inflammation, mucus hypersecretion and their possible mechanism in asthma mice. BALB/c mice were sensitized and challenged with ovalbumin (OVA) for establishment of asthma model and then treated with NBP during day 22-77. The pulmonary function of the mice was determined, and the pathology of lung tissue and goblet cell hyperplasia were observed through analyzing inflammation scores and goblet cell percentage, respectively. Cytokine IL-4, IL-8, IL-13 and tumor necrosis factor-alpha (TNF-α) in bronchoalveolar lavage fluid (BALF) and total immunogloblin E (T-IgE) and OVA-specific IgE in serum were examined by enzyme-linked immunosorbent assay (ELISA). The expressions of Mucin 5AC (Muc5ac) and nuclear transcription factor-kappa B (NF-κB) in lung tissues were evaluated by immunohistochemistry, western blot and real-time polymerase chain reaction (RT-PCR). The results show that 50 mg/kg NBP significantly reduced OVA-induced increase in inflammation scoring, goblet cell percentage and mucus secretion of airway tissue, and improved the pulmonary function. NBP could also decrease IL-4, IL-8 IL-13, and TNF-α in BALF and T-IgE and OVA-specific IgE in serum. The expression of Muc5ac and NF-κB in lung tissue was significantly down-regulated after NBP treatment. This study suggested that NBP may effectively inhibit airway inflammation and mucus hypersecretion in asthma by modulating NF-κB activation.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Benzofurans; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Leukocyte Count; Lung; Mice, Inbred BALB C; Mucin 5AC; Mucus; NF-kappa B; Ovalbumin

Rats were divided into six groups: sham/control , Dl-3-n-butylphthalide, P1 (low phenytoin, 100 mg/kg), P2 (high phenytoin, 200 mg/kg), NP1 (Dl-3-n-butylphthalide 80 mg/kg, phenytoin 100 mg/kg), NP2 (Dl-3-n-butylphthalide 80 mg/kg, phenytoin 200 mg/kg). Hematoxylin/eosin and Nissl staining showed that, compared to the sham/control group, the Dl-3-n-butylphthalide group had no obvious hippocampal and cerebellar neuron loss, but there was a significant neuron loss in the P1 and P2 groups (P < 0.05), which was more obvious in the P2 group (P < 0.05). The positive expression of Bax and Bcl-2 proteins in hippocampal and cerebellar neurons was not significantly different between sham and Dl-3-n-butylphthalide groups; however, compared to sham, Bax expression was significantly increased and Bcl-2 was significantly decreased in the hippocampal and cerebellar neurons of rats in both P1 and P2 groups (P < 0.05), being more obvious in the P2 group (P < 0.05). Furthermore, the administration of Dl-3-n-butylphthalide attenuated the deleterious effects of phenytoin (P < 0.05). Our results indicate that phenytoin causes apoptosis of hippocampal and cerebellar neurons in rats in a dose-dependent manner, with the effect of a higher dose being more obvious, whereas, Dl-3-n-butylphthalide inhibits the phenytoin-induced apoptosis of neurons and has a neuroprotective role.

Topics: Animals; Anticonvulsants; Apoptosis; Benzofurans; Cerebellum; Hippocampus; Male; Neurons; Neuroprotective Agents; Phenytoin; Rats; Rats, Sprague-Dawley

The aim of this study was to investigate the effects of butylphthalide on oxidative stress and inflammatory response in rats with myocardial infarction through the protein kinase B/nuclear factor E2-related factor 2 (Akt/Nrf2) signaling pathway.. A total of 36 Sprague-Dawley rats were randomly divided into three groups, including sham group (n=12), model group (n=12) and butylphthalide group (n=12). In the sham group, the heart was exposed, and normal saline was intraperitoneally injected after the operation. In the model group, acute myocardial infarction (AMI) model was established, and normal saline was intraperitoneally injected after the operation. In the butylphthalide group, AMI model was established, and butylphthalide was intraperitoneally injected after the operation. After intervention for 4 weeks, the rats were killed, and the samples were collected. The morphology of heart tissues was observed via hematoxylin-eosin (HE) staining. The expression of nicotinamide adenine dinucleotide phosphate oxidase-1 (NOX-1) was detected via immunohistochemistry. The protein expressions of phosphorylated Akt (p-Akt) and p-Nrf2 were detected via Western blotting. Moreover, the content of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) was detected via enzyme-linked immunosorbent assay (ELISA). The messenger ribonucleic acid (mRNA) expression levels of IL-1β, TNF-α and NOX-1 were detected via quantitative Polymerase Chain reaction (qPCR). Furthermore, the apoptosis rate of the cells was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.. The morphology of heart tissues was significantly damaged in the model group and butylphthalide group compared with the sham group. However, it was significantly improved in the butylphthalide group when compared with the model group. The expression level of NOX-1 increased markedly in the model group and butylphthalide group compared with the sham group (p<0.05). However, it was remarkably reduced in the butylphthalide group compared with the model group (p<0.05). Meanwhile, the protein expression levels of p-Akt and p-Nrf2 were significantly higher in the model group and butylphthalide group than those of the sham group (p<0.05). However, the protein expression levels of p-Akt and p-Nrf2 in the butylphthalide group were remarkably lower than the model group (p<0.05). The mRNA expression levels of IL-1β, TNF-α and NOX-1 were markedly higher in the model group and butylphthalide group than those of the sham group (p<0.05). However, they remarkably declined in the butylphthalide group compared with the model group (p<0.05). In addition, the content of IL-1β and TNF-α increased in the model group and butylphthalide group when compared with the sham group (p<0.05). The content of IL-1β and TNF-α in the butylphthalide group was significantly lower than the model group (p<0.05). Furthermore, the apoptosis rate was significantly higher in the model group and butylphthalide group than the sham group (p<0.05), which was significantly lower in the butylphthalide group than the model group (p<0.05).. Butylphthalide inhibits inflammatory and oxidative stress responses after AMI by regulating the Akt/Nrf2 signaling pathway, thereby inhibiting myocardial apoptosis and benefiting the morphological repair of myocardial tissues.

Topics: Animals; Benzofurans; Disease Models, Animal; Inflammation; Male; Myocardial Infarction; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidative Stress; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction

To investigate the alteration in Golgi and blood-brain barrier after cerebral hemorrhage in SD rats, and to evaluate the effect of butylphthalide on blood-brain barrier. 
 Methods: Sprague-Dawley rats were randomly distributed into 4 groups: a control group, a sham group, an intracerebral hemorrhage (ICH) group, and a butylphthalide group. Brain tissue was collected at 48 h after the blood brain barrier permeability was examined. Western blotting and real-time polymerase chain reaction (real-time PCR) were conducted to explore the change of GM130, Cdc42 and tight junction protein and mRNA expression in rat brain after ICH. Immunohistochemistry (IHC) was performed to explore the distribution of ZO-1 and Occludin in the cerebral vascular endothelial cells around the hematoma.
 Results: The Evans blue (EB) extravasation in the ICH group were much greater than that in the sham group (P<0.05). Butylphthalide treatment significantly decreased Evans blue extravasation compared to the ICH group (P<0.05). Results of Western blotting and real-time PCR showed that GM130, Cdc42, ZO-1/Occludin were decreased (P<0.05). The intervention of butylphthalide significantly upregulated the expressions of Cdc42 as well as ZO-1/Occludin (P<0.05), but exerted no effect on GM130 (P<0.05). Immunofluorescent staining showed that GM130 was co-localized with Cdc42 and administration of butylphthalide improved the expression of Cdc42 around the hematoma without affecting the expression of GM130. IHC showed that expressions of occludin and ZO-1 around the hematoma were significantly decreased in the ICH group (P<0.05), whereas butylphthalide treatment elevated the expressions of ZO-1 and occludin around the hematoma compared with the ICH group (P<0.05).
 Conclusion: Morphology of Golgi apparatus is altered and the blood-brain barrier is destroyed after ICH. The application of butylphthalide can alleviate neurological impairment and blood-brain barrier disruption, which is related to the up-regulation of Cdc42, but not GM130.. 目的：探讨SD大鼠脑出血后高尔基体和血脑屏障的变化及丁苯酞的干预作用和机制。方法：SD大鼠随机分为正常对照组、假手术组、脑出血组及丁苯酞组。造模48 h后测量各组的伊文思蓝含量，采用蛋白质印迹法及real-time PCR检测Cdc42，GM130，ZO-1和Occludin蛋白灰度比值及mRNA相对表达量，免疫荧光观察Cdc42和GM130是否存在共定位及脑出血后丁苯酞干预的荧光强度，免疫组织化学观察ZO-1和Occludin在血肿周围内皮细胞上的分布。结果：脑出血组的神经功能评分及伊文思蓝含量明显高于假手术组(P<0.05)；丁苯酞组的神经功能评分及伊文思蓝含量较脑出血组明显降低(P<0.05)。 脑出血组Occludin，ZO-1，GM130及Cdc42蛋白灰度比值及mRNA相对表达量较假手术组降低(P<0.05)；丁苯酞组中Occludin，ZO-1及Cdc42蛋白灰度比值及mRNA相对表达量较脑出血组升高(P<0.05)，而GM130 mRNA差异无统计学意义(P>0.05)。免疫组织化学发现脑出血组中Occludin和ZO-1在血肿周围的表达较假手术组降低(P<0.05)，而丁苯酞组则增加(P<0.05)。结论：脑出血后高尔基体形态发生改变，血脑屏障受到破坏；丁苯酞能减轻神经功能缺损及血脑屏障破坏程度，其机制与Cdc42的上调相关，而与GM130可能无关。.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Cerebral Hemorrhage; Endothelial Cells; Rats; Rats, Sprague-Dawley

Oxidative stress and neuroinflammation play important roles in the pathology of Alzheimer's disease (AD). Thioredoxin-interacting protein (TXNIP), an endogenous inhibitor of antioxidant thioredoxin, is suspected to be an important modulator of oxidative stress and inflammation. However, the underlying mechanism involved in the abnormal homeostasis of TXNIP-thioredoxin (TrX) in AD pathogenesis remains unclear.. Using the Swedish mutant form of APP (APPswe)/PSEN1dE9 transgenic mouse (APP/PS1) and human-derived neuronal cells as model systems, we disclosed the impairment of the nuclear factor erythroid 2-related factor 2 (Nrf2)-TXNIP-TrX signaling in Alzheimer's-like pathology. We observed that the immune staining of TXNIP was increased in postmortem AD brain. The chronic accumulation of inflammatory mediator in neuronal cells facilitates interactions of TXNIP-nucleotide binding oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3) and NLRP3-ASC, which increases β-amyloid (Aβ) secretion. The antioxidant Dl-3-n-butylphthalide (Dl-NBP) is commonly used for cerebral ischemia treatment. In our study, we elucidated for new mechanisms by which Dl-NBP enhanced TrX activity, suppressed TXNIP, and ameliorated neuronal apoptosis in the APP/PS1 mouse brains. In human glioblastoma A172 cells and neuroblastoma SH-SY5Y cells, we delineated the Dl-NBP-mediated signaling pathways by which Dl-NBP-dependent upregulation of Nrf2 mediated the reciprocal regulation of reducing proinflammatory cytokine and inhibiting Aβ production in the glial and neuronal cells overexpressing APPswe.. Our data provide a novel insight into the molecular mechanism that impairments of Nrf2-TXNIP-TrX system may be involved in the imbalance of cellular redox homeostasis and inflammatory damage in the AD brain.. Dl-NBP treatment could suppress TXNIP-NLRP3 interaction and inhibit NLRP3 inflammasome activation via upregulating Nrf2. These findings may provide an instrumental therapeutic approach for AD. Antioxid. Redox Signal. 00, 000-000.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Amyloid Precursor Protein Secretases; Animals; Antioxidants; Apoptosis; Benzofurans; Carrier Proteins; Cell Line, Tumor; HEK293 Cells; Humans; Inflammasomes; Interleukin-1beta; Male; Mice; Mice, Transgenic; NF-E2-Related Factor 2; NLR Family, Pyrin Domain-Containing 3 Protein; Protein Binding; Thioredoxins

Dl-3-N-butylphthalide (NBP) is approved in China for the treatment of ischemic stroke. Previous studies have shown that NBP promotes recovery after stroke via multiple mechanisms. However, the effect of NBP on vascular function and thrombosis remains unclear. Here, we aim to study the effect of NBP on vascular function using a rat model of transient middle cerebral artery occlusion (MCAO) and a state-of-the-art high-resolution synchrotron radiation angiography. Eighty SD rats underwent MCAO surgery. NBP (90 mg/kg) was administrated daily by gavage. Synchrotron radiation angiography was used to evaluate the cerebral vascular perfusion, vasoconstriction, and vasodilation in real-time. Neurological scores, brain infarction and atrophy were evaluated. Real-time PCR was used to assess the expression levels of thrombosis and vasoconstriction-related genes. Results revealed that NBP attenuated thrombosis after MCAO and reduced brain infarct and atrophy volume. NBP administrated at 1 and 4 h after MCAO prevented the vasoconstriction of the artery and maintained its diameter at normal level. Administrated at one week after surgery, NBP functioned as a vasodilator in rats after MCAO while displayed no vasodilating effect in sham group. Our results suggested that NBP attenuates brain injury via increasing the regional blood flow by reducing thrombosis and vasoconstriction.

Topics: Animals; Benzofurans; Cerebral Arteries; Cerebrovascular Circulation; Infarction, Middle Cerebral Artery; Male; Neuroprotective Agents; Platelet Aggregation Inhibitors; Rats, Sprague-Dawley; Reperfusion Injury; Vasodilator Agents

Drug-drug interactions (DDIs) are thought to be associated with the inhibition of cytochrome P450 activities. The cocktail method with analysis of the metabolism of two or more probe drugs is used to determine CYP450 activities. In this study, we established a UHPLC-MS/MS method for simultaneous quantitation of four CYP450 probe drugs (phenacetin, omeprazole, metoprolol and midazolam) and their metabolites (acetaminophen, 5'-hydroxy omeprazole, α-hydroxy metoprolol and 1'-hydroxy midazolam) in rat plasma. Sample preparation by plasma protein precipitation was combined with a liquid-liquid extraction method. The separation was carried out on a ZORBAX Eclipse Plus C18 Rapid Resolution High Definition column with a gradient elution, using water containing 0.1% formic acid (A) and acetonitrile (B) in a run time of only 3.0 min. Detection was conducted with a 6420 series triple-quadrupole tandem mass spectrometer, using ESI in positive ion mode with multiple reaction monitoring (MRM). The calibration curves were linear over the concentration range 10-5000 ng/mL for phenacetin, omeprazole, metoprolol and midazolam, and 1-500 ng/mL for their metabolites. Intra- and inter-day precisions were within 15%, and the accuracies were in the range of 87-112%. The method was successfully applied to the pharmacokinetic study of probe drugs/metabolites and DDIs with 3-n-butylphthalide (NBP) after administration of a single oral dose of phenacetin, omeprazole, metoprolol and midazolam in rats.

Topics: Administration, Oral; Animals; Benzofurans; Chromatography, High Pressure Liquid; Cytochrome P-450 Enzyme System; Drug Interactions; Liquid-Liquid Extraction; Male; Metoprolol; Midazolam; Omeprazole; Phenacetin; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Tandem Mass Spectrometry

Numerous epidemiological studies have shown that diabetes mellitus (DM) is associated with dementia and cognition decline. However, there is currently no effective treatment for diabetes-induced cognitive dysfunction. The neuroprotective effect of L-3-n-butylphthalide (L-NBP) has been demonstrated in vascular dementia animal models. The purpose of this study was to determine whether L-NBP can ameliorate cognitive deficits in db/db mice, a model of obesity and type 2 diabetes. The mice were administered with vehicle or L-NBP (120 mg/kg) by gavage daily for 6 weeks. Then, Morris water maze tasks were performed, and hippocampal LTP was recorded in vivo. Next, the synaptic structure of the CA1 hippocampus region was investigated via electron microscopy. Finally, the expression levels of MDA, SOD, 8-OHdG, and NADPH oxidase subunits gp91 and p67, as well as the expression of NF-κB p65, TNF-α, IL-1β and caspase-3 were measured by Western blot, RT-PCR and ELISA. Treatment with L-NBP significantly attenuated the learning and memory deficits in db/db mice. Concomitantly, L-NBP also increased hippocampus synaptic plasticity, characterized by an enhanced in vivo LTP, and suppressed oxidative stress, as indicated by increased SOD activity and decreased MDA, 8-OHdG, and NADPH oxidase subunits p67 and gp91. L-NBP also significantly decreased NF-κB p65, TNF-α, IL-1βand caspase-3 levels in the hippocampus. L-NBP significantly ameliorated cognitive decline in type 2 diabetic mice, and this effect was accompanied by an improvement in hippocampal plasticity and an amelioration of oxidative stress, inflammation and apoptosis cascades. Thus, L-NBP may be a promising therapeutic agent against DM-mediated cognitive dysfunction.

Topics: Animals; Benzofurans; Caspase 3; Cognition; Cognitive Dysfunction; Cytokines; Diabetes Mellitus, Type 2; Disease Models, Animal; Hippocampus; Maze Learning; Mice; Neuroprotective Agents; Oxidative Stress

Seven new butylphthalide derivatives, ligusticumolide A-G (1-7), together with two known butylphthalide derivatives (8-9) were isolated from an ethanol extract of Ligusticum chuanxiong Hort. The structures of these derivatives were elucidated from analysis of 1D/2D NMR, UV, IR and HRESIMS data. The absolute configurations of these derivatives were determined by electronic circular dichroism (ECD) calculations and Mosher's method. Ligusticumolide A (1) and ligusticumolide B (2) are enantiomers that were obtained by chiral separation. Ligusticumolide C (3) and ligusticumolide D (4) are diastereomers. All of the compounds were evaluated for their hepatoprotective activity against N-acetyl-p-aminophenol-induced HepG2 cell injury. Compounds 4, 5, and 7-9 showed more significant hepatoprotective activity than that of the positive control drug (bicyclol) at a concentration of 10 μM (p < 0.01).

Topics: Benzofurans; Cell Line, Tumor; Cell Survival; Circular Dichroism; Humans; Ligusticum; Magnetic Resonance Spectroscopy; Molecular Conformation; Plant Roots; Protective Agents; Stereoisomerism

In China, L-3-n-butylphthalide (L-NBP) showed promising pharmacological actions in stroke treatment. Analyzing the characteristics of L-NBP might provide valuable hints for new drug design. The current study is aimed to determine the effects of L-NBP on neuritogenesis and further to elucidate the neuronal protection against stroke impairment in vitro. L-NBP was applied to rat pheochromocytoma PC12 cells and cultured rat cortical neurons under the normoxic condition and the oxygen-glucose deprivation/reoxygenation (OGD/R) insults, respectively. Immunofluorescence staining, western blot analysis, Sholl analysis, lactate dehydrogenase (LDH) release assay, 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) reduction assay and enzyme-linked immunosorbent assay (ELISA) were performed. L-NBP could concentration-dependently stimulate the development of growth cones, enhance the neuritic branches and synapse formation. It indicated that L-NBP possibly promoted the neuritogenic activity in a stage-dependent manner. Further research proved that L-NBP could promptly activate epidermal growth factor (EGF) receptor, up-regulate the expressions of extracellular signal-regulated kinase1/2 (ERK1/2), cAMP response element-binding protein (CREB) and E-26-like protein 1 (ELK-1). In addition, L-NBP enhanced the sustained expressions of brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF). The inhibition to the receptors of EGF, NGF,BDNF could attenuate L-NBP induced neuritogenic and neuronal survival after the OGD/R toxicity. Basing on these investigations, we concluded that L-NBP might reconstruct the impaired neuronal network and improved the neuronal complexity after the ischemic insults through multiple pathways which at least were via the activations of EGF receptor, BDNF and NGF related signals.

Topics: Animals; Benzofurans; Cell Hypoxia; Cell Survival; Cells, Cultured; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Neurites; Neurogenesis; Neurons; Neuroprotective Agents; PC12 Cells; Rats; Rats, Sprague-Dawley

Postoperative neurological deficits impair the overall outcome of revascularization surgery for patients with moyamoya disease (MMD). dl-3-n-butylphthalide (NBP) is approved for the treatment of ischemic stroke in China. This pilot study evaluated the effect of NBP on perioperative stroke and neurological deficits in patients with MMD.. The authors studied cases in which patients underwent combined revascularization surgery for MMD at their institution, with or without NBP administration. The overall study group included 164 patients (213 surgically treated hemispheres), including 49 patients who received NBP (25 mg twice daily) for 7 postoperative days. The incidence of perioperative stroke and transient neurological deficit (TND) and the severity of neurological deficits were compared between 49 propensity score-matched case pairs with or without NBP treatment. Subgroup analyses by type of onset and preoperative neurological status were also performed to determine specific characteristics of patients who might benefit from NBP administration.. In the overall cohort, baseline characteristics differed with respect to preoperative stroke and modified Rankin Scale (mRS) score between patients who received NBP and those who did not receive it. In the 49 propensity score-matched pairs, postoperative stroke was observed in 11 patients and TND occurred in 21 patients, with no significant difference in incidence between the 2 groups. However, the TND was less severe in the NBP-treated group (p = 0.01). At 1 month after surgery, the neurological outcome was more favorable (p = 0.001) and the disability-free recovery rate was higher in patients with NBP treatment (p < 0.001). The number of patients who experienced an improved neurological function, compared to preoperative function, as measured by mRS, was greater in the NBP group than in the no-NBP group (p < 0.001). Multivariable analysis revealed that NBP administration was associated with decreased severity of TND (OR 0.28, p = 0.02), improved neurological function (OR 65.29, p = 0.04), and lower postoperative mRS score (OR 0.06, p < 0.001). These beneficial effects of NBP remained significant in ischemic type MMD and patients with preoperative mRS scores of 2 or greater.. Postoperative administration of NBP may alleviate perioperative neurological deficits after revascularization surgery for MMD, especially in patients with ischemic MMD and unfavorable preoperative status. The results of this study suggest that randomized controlled trials to assess the potential benefit of NBP in patients with MMD may be warranted.

Topics: Adult; Benzofurans; Cerebral Revascularization; Cohort Studies; Combined Modality Therapy; Female; Humans; Male; Middle Aged; Moyamoya Disease; Nervous System Diseases; Neuroprotective Agents; Pilot Projects; Postoperative Complications; Propensity Score; Prospective Studies

Topics: Aged; Benzofurans; Continuous Positive Airway Pressure; Endothelin-1; Humans; Neuroprotective Agents; Nitric Oxide; Sleep Apnea, Obstructive

Previous findings have demonstrated, in a rat model, that chronic cerebral hypoperfusion (CCH) decreases cortical cerebral blood flow (CBF) while Dl-3-n-Butylphthalide (DNB) accelerates the timely recovery of CBF. However, potential biomarkers, therapeutic targets, and underlying mechanisms for these processes are unclear. In this study, a solid-phase antibody microarray for simultaneously detecting multiple proteins was used to search cortex biotargets in CCH compared to a sham control group, and these results were further examined by biological functional analysis. After DNB treatment, western blot and immunostaining were used to verify candidate protein expression. Importantly, we identified seven proteins that may serve as novel biotargets contributing to CCH. The levels of Tie-2, CNTFRα, IL-4, IL-10, ITGAM, MDC, and TROY were uniquely altered in the CCH. The Tie-2 level was significantly decreased and identified in CCH 2 week (W), CCH 4 W and CCH 8 W. In addition, Ang-1 level and Ang-1/Ang-2 ratio were significantly decreased in CCH 2 W and CCH 4 W while Ang-2 level was increased in the CCH, whereas DNB treatment created the inverse effect to some extent. Moreover, the expression of VEGF and CD34 in the earlier stage of CCH and the diameters of bilateral vertebral arteries (VAs), were significantly enlarged by DNB treatment. Together, we found that the Ang-1/Ang-2/Tie-2 signaling axis was altered in the CCH rat cortex, and DNB treatment could timely regulate this angiopoietin/Tie signaling axis to promote neovascularization in early stages.

Topics: Angiopoietin-1; Angiopoietin-2; Animals; Benzofurans; Blotting, Western; Brain Ischemia; Cerebral Cortex; Cerebrovascular Circulation; Male; Neovascularization, Physiologic; Protein Array Analysis; Rats; Rats, Sprague-Dawley; Receptor, TIE-2; Signal Transduction

The increased permeability of the blood-brain barrier (BBB) induced by ischemia/hypoxia is generally correlated with alteration of tight junctions (TJs). DL-3-n-butylphthalide (NBP) has been shown to exert neuroprotective effects after ischemic injury. However, few studies have assessed the correlation between NBP and TJs. This study aimed to investigate the potential effect of NBP on the TJ proteins claudin-5, zonula occludens-1 (ZO-1), and occludin during brain ischemia.. A chronic cerebral hypoperfusion (CCH) Sprague-Dawley rat model was established, and NBP (20, 40, or 80 mg/kg, gavage, once a day) treatment was performed for 14 days. NBP (0.1 or 1.0 μmol/L) pre-treatment was applied to an in vitro hypoxia microvascular endothelial cell model (1% O2, 24 h). BBB permeability was assessed by performing the Evans blue assay. The expressions and localization of claudin-5, ZO-1, occludin, phosphorylated/total protein kinase B (p-Akt/Akt), phosphorylated/total glycogen synthase kinase 3β (GSK-3β)/GSK-3β, and β-catenin/β-actin were evaluated by Western blotting or immunofluorescence. Reactive oxygen species (ROS) generation was measured by flow cytometry analysis. TJ ultrastructure was observed by transmission electron microscopy.. In CCH rats, treatment with 40 and 80 mg/kg NBP decreased the Evans blue content in brain tissue (9.0 ± 0.9 μg/g vs. 12.3 ± 1.9 μg/g, P = 0.005; 6.7 ± 0.6 μg/g vs. 12.3 ± 1.9 μg/g, P < 0.01), increased the expression of claudin-5 (0.79 ± 0.08 vs. 0.41 ± 0.06, P < 0.01; 0.97 ± 0.07 vs. 0.41 ± 0.06, P < 0.01), and elevated the ZO-1 protein level (P < 0.05) in brain microvascular segments in a dose-dependent manner in comparison with the corresponding values in the model group. There was no significant difference in occludin expression (P > 0.05). In the hypoxia cell model, NBP pre-treatment improved TJ ultrastructure, decreased intracellular ROS level, and increased the expression of claudin-5 (P < 0.01) and ZO-1 (P < 0.01) in comparison with the corresponding values in the hypoxia group. NBP treatment also elevated the relative expression levels of p-Akt/Akt, p-GSK-3β/GSK-3β, and β-catenin/β-actin in comparison with the corresponding values in the hypoxia group (all P < 0.05).. NBP improves the barrier function of BBB against ischemic injury by upregulating the expression of TJ proteins, possibly by reducing oxidative stress and activating the Akt/GSK-3β/β-catenin signaling pathway.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Blotting, Western; Brain Ischemia; Cells, Cultured; Claudin-5; Immunohistochemistry; Male; Microscopy, Electron, Transmission; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Tight Junction Proteins; Zonula Occludens-1 Protein

Remyelination has been widely noticed as an important repair mechanism triggered after a stroke-induced white matter injury, but it often fails due to the lack of recruitment of the oligodendrocyte progenitor cells (OPCs) to the demyelinated area and the inadequate differentiation of OPCs. Racemic dl-3-n-butylphthalide (dl-NBP) has been reported to improve the functional recovery in animal models of vascular dementia, Alzheimer's disease (AD) and ischemic stroke. Dl-NBP (70 mg/kg) by oral gavage for two weeks from day 7 after a stroke was administered in the study, the treatment promoted differentiation and maturation of OPCs in perilesional white matter and enhanced the length of crossing corticospinal tract (CST) fibers into the denervated hemispheres. These effects could be linked to increased expression levels of brain-derived neurotrophic factor (BDNF) and the reduced expression of neurite outgrowth inhibitor (NogoA) in the perilesional area in dl-NBP group. However, dl-NBP did not increase the numbers of neuron/glia type 2 (NG2)-positive and oligodendrocyte lineage transcription factor 2 (Olig2)-positive cells in the subventricular zone. Our data highlight the effects of dl-NBP in the remyelination process and reveal the therapeutic potential of this approach in cerebral ischemia.

Topics: Animals; Benzofurans; Brain Ischemia; Brain-Derived Neurotrophic Factor; Cell Differentiation; Male; Myelin Sheath; Nogo Proteins; Oligodendrocyte Precursor Cells; Oligodendroglia; Rats; Rats, Wistar; Recovery of Function; Remyelination; Stroke; White Matter

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS). The disease mechanisms driving progressive MS remain unresolved. Without this information, current therapeutic strategies are unsatisfactory in preventing disease progression. Our previous work revealed that DL-3-n-butylphthalide (NBP) treatment reduced demyelination in an ethidium bromide mouse model of demyelination. Here, we examine the effect of NBP in the cuprizone model of demyelination by evaluating the pathologic, functional, and behavioral consequences of treatment with NBP.. Forty mice were divided randomly into 4 groups: a normal diet group, a cuprizone diet group, and two NBP groups (10 and 20 mg/kg). CNS infiltration by microglia, axon health and myelination were assessed using immunohistochemistry and electron microscopy, and the levels of cytoplasmic complexes were assessed by Western blotting.. The results showed the neuroprotective effects of the NBP included suppressing the microglia activation through inhibition of nuclear factor-κB (NF-κB) expression, thus decreasing activation of the NF-κB signaling pathway. In particular, myelin density was increased due to an increased mean number of mature oligodendrocytes (OLs) in the high-dose NBP (20 mg/kg) subgroup through reduced oligodendrocyte apoptosis. Meanwhile, increased expression of myelin sheath proteins, including proteolipid protein (PLP) and myelin basic protein (MBP), was observed in the same subgroup.. These data suggest that NBP may not only have anti-inflammatory properties but also promote the survival of OLs in a mouse cuprizone model of demyelination. NBP may have a potential role in the treatment of MS.

Topics: Animals; Astrocytes; Axons; Benzofurans; Corpus Callosum; Cuprizone; Demyelinating Diseases; Disease Models, Animal; Gene Expression Regulation; Male; Mice; Mice, Inbred C57BL; Microglia; Multiple Sclerosis; Myelin Sheath; Neuroprotective Agents; NF-kappa B; Oligodendroglia; Signal Transduction

The aim of this study was to investigate the effect of 3-n-butylphthalide (NBP) on the apoptosis of nerve cells in vascular dementia (VaD) model rats caused by cerebral small vessel disease (CSVD), and to explore its regulatory mechanism.. The model of VaD was successfully established in rats by carotid artery ligation. All rats were randomly divided into three groups, including the sham operation group, model group and NBP group. The neurobehavioral score was used to verify whether the model was successfully established. The changes in learning and memory abilities of rats were detected via water maze experiment. The levels of Bcl-2-associated X protein (Bax) and cysteinyl aspartate specific proteinase-3 (Caspase-3) in the serum of rats was detected by enzyme-linked immunosorbent assay (ELISA). Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was adopted to detect the apoptosis of nerve cells in brain tissues of rats. Moreover, the protein levels of phosphorylated phosphatidylinositol-3-kinase (PI3K) and phosphorylated protein kinase B (Akt) in brain tissues of rats were measured using Western blotting.. Compared with the sham operation group, the neurobehavioral score of rats increased significantly, whereas learning and memory abilities decreased markedly in the model group. The levels of Bax and Caspase-3 in rat serum were remarkably up-regulated, and the apoptosis rate of nerve cells in brain tissues of rats increased significantly in the model group as well. Meanwhile, the levels of phosphorylated PI3K and phosphorylated Akt were notably declined. Compared with the model group, the neurobehavioral score decreased markedly, while learning and memory abilities were remarkably improved in the NBP group. The levels of Bax and Caspase-3 in rat serum were significantly down-regulated, and the apoptosis rate of nerve cells in brain tissues of rats were reduced in the NBP group. Furthermore, the protein levels of phosphorylated PI3K and phosphorylated Akt were remarkably elevated in the NBP group.. NBP can improve the morphology of brain tissue cells and the learning and memory abilities, and inhibit the apoptosis of nerve cells in VaD model rats with CSVD. The possible underlying mechanism may be related to the activation of the PI3K/Akt signaling pathway.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Behavior, Animal; Benzofurans; Brain Chemistry; Carotid Artery Diseases; Caspase 3; Cerebral Small Vessel Diseases; Male; Maze Learning; Neurons; Neuroprotective Agents; Oncogene Protein v-akt; Phosphatidylinositol 3-Kinases; Rats; Rats, Sprague-Dawley; Signal Transduction

Animal experiments verified that dl-3-n-butylphthalide (NBP) can protect vascular endothelial cells from ischemic damage and promote vascular proliferation in ischemic stroke treatment, but the underlying mechanism has not been fully clarified. This study aimed to investigate the effects of NBP on peroxisome proliferators-activated receptor-γ coactivator-1α (PGC-1α) expression in endothelial cells exposed to oxygen-glucose deprivation (OGD) and to clarify the related molecular mechanism.. SV40-transformed aortic rat endothelial cell line was cultured and subjected to OGD in the presence or absence of NBP. The cell viability was evaluated by using thiazolyl blue tetrazolium bromide (MTT) method. The cellular endothelial nitric oxide synthase (eNOS) activity was measured by using eNOS activity assay. The nuclear changes were assessed with Hoechst 33342 fluorescent dye. The immunofluorescence analysis and Western blotting assay were conducted to evaluate the protein expression.. We found that NBP could significantly prevent endothelial cells from OGD-induced injuries, in terms of cell morphology and cell viability. Both immunofluorescence analysis and Western blot findings confirmed that the NBP treatment further enhanced PGC-1α expression during OGD, which was prevented in the presence of selective endothelial nitric oxide synthetase (eNOS) inhibitor N5-(1-Iminoethyl)-L-ornithine-HCL (L-NIO). Furthermore, we found that NBP could protect the eNOS activity about by 40% during OGD and did not influence the eNOS protein level in the spectrophotometric-based analysis.. NBP maintained the endothelial PGC-1α expression via regulating eNOS activity during the exposure to OGD; therefore, it presented its protective function to cell viability and vascular proliferation.

Topics: Benzofurans; Cell Hypoxia; Cell Line; Cell Shape; Cell Survival; Endothelial Cells; Enzyme Inhibitors; Glucose; Humans; Neuroprotective Agents; Nitric Oxide Synthase Type III; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha

The acute respiratory distress syndrome (ARDS), a devastating clinical syndrome, is one of the most severe complications of acute lung injury (ALI). Despite of decades of clinical trials and supportive ventilation strategies, the incidence and mortality of ALI/ARDS remain high. DL-3-n-butylphthalide (NBP) is a synthesized raceme of L-3-n-butylphthalide which has been approved to possess various activities. In the current study, we aimed to investigate the effect of NBP on ALI in lipopolysaccharide (LPS)-treated mice. We found that 10 mg/kg and 50 mg/kg NBP significantly prevented LPS-induced increase of W/D ratio of lung, histological injury of lung, infiltration of inflammatory cells, release of pro-inflammatory cytokines and chemokines, and oxidative damage. Sirtuin 1 (SIRT1) and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) expression was increased by NBP in lung of LPS-treated mice. Knockout of SIRT1 significantly aggravated LPS-induced ALI. Moreover, the absence of SIRT1 notably inhibited NBP-induced protective effects against LPS-induced increase of W/D ratio of lung, histological injury of lung, infiltration of inflammatory cells, release of pro-inflammatory cytokines and chemokines, and oxidative damage. However, knockout of SIRT1 did not completely inhibit NBP-induced upregulation of Nrf2 and attenuation of ALI. The results demonstrated that NBP could activate Nrf2 antioxidant signaling in a SIRT1-dependent and SIRT1-independent manner, resulting in the amelioration of oxidative stress, inflammation and pulmonary edema. The data highlights the importance of SIRT1/Nrf2 signaling in the protective effects of NBP.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Antioxidants; Benzofurans; Bronchoalveolar Lavage Fluid; Cytokines; Lipopolysaccharides; Lung; Male; Mice, Inbred C57BL; Mice, Knockout; NF-E2-Related Factor 2; Pulmonary Edema; Sirtuin 1

Topics: Apoptosis; Benzofurans; Blotting, Western; Cell Line; Cell Survival; Endoplasmic Reticulum; Ependymoglial Cells; Fluoresceins; Fluorescent Antibody Technique; Heme Oxygenase-1; Humans; Hydrogen Peroxide; Membrane Potential, Mitochondrial; Microscopy, Fluorescence; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidants; Oxidative Stress; Reactive Oxygen Species; Retinal Diseases; Signal Transduction

Topics: 4-Butyrolactone; Administration, Oral; Animals; Benzofurans; Drugs, Chinese Herbal; Gas Chromatography-Mass Spectrometry; Limit of Detection; Linear Models; Male; Rats; Rats, Sprague-Dawley; Reproducibility of Results

Ischemic stroke occurs following arterial occlusion and subsequent blood flow cease, and restoration of blood supply by thrombolytic therapy may cause cerebral ischemic reperfusion (IR) injury resulting in breakdowns of blood-brain barrier (BBB). Dl-3-n-butylphthalide (NBP) is an extraction from Chinese celery Apium graveolens Linn seeds and has neuroprotective effects in ischemic stroke. This study explored effects of NBP on BBB disruption caused by cerebral IR and transformation of tight junctions (TJs)-associated proteins and caveolae. Our results demonstrated that NBP alleviated cerebral IR-induced deterioration of vascular permeability by up-regulating TJ-associated proteins but down-regulating caveolin-1. NBP significantly improved neurological function and cerebral blood flow but reduced cerebral edema and infarct volume after IR. In conclusion, NBP exerts neuroprotective effects through attenuating cerebral infarct volume and neurological deficit score, reducing cerebral edema and BBB permeability. The neuroprotective effect of NBP is possibly related to its ability to improve blood flow in cerebral ischemic areas. NBP may turn into a novel treatment drug to prevent BBB dysfunction in ischemic stroke.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Brain Edema; Brain Ischemia; Capillary Permeability; Caveolin 1; Cerebrovascular Circulation; Disease Models, Animal; Male; Mice; Neuroprotective Agents; Random Allocation; Reperfusion Injury; Tight Junctions

Effects of dl-3-n-butylphthalide (NBP) on white matter damage and cognitive impairment in vascular cognitive impairment (VCI) have not been well studied. This study aimed to investigate the effects of NBP treatment on chronic cerebral hypoperfusion-induced white matter lesions and cognitive dysfunction in mice.. Mice were subjected to bilateral common carotid artery stenosis (BCAS) for over 30 days. The cerebral blood flow was detected using a laser Doppler flowmetry. Cognitive functions were assessed by several behavioral tests. We also evaluated the effects of NBP on the blood-brain barrier (BBB) disruption and reactive astrogliosis, using Evans Blue extravasation, Western blot, CBA, and immunofluorescence in BCAS mice and cultured astrocytes.. The results indicated that NBP treatment attenuated spatial memory dysfunction while promoted cerebral perfusion and white matter integrity in BCAS mice. Moreover, NBP treatment prevented BBB leakage and damage of endothelial cells, as well as disruption of endothelial tight junctions. Furthermore, NBP administration effectively decreased the number of activated astrocytes and pro-inflammatory cytokines, as well as the production of MMPs, in BCAS-induced mice and LPS-stimulated astrocytes.. Our results indicated that NBP represents a promising therapy for chronic cerebral hypoperfusion-induced white matter damage and cognitive impairment.

Topics: Animals; Benzofurans; Brain Ischemia; Carotid Stenosis; Cells, Cultured; Cerebrovascular Circulation; Chronic Disease; Cognitive Dysfunction; Male; Mice; Mice, Inbred C57BL; Neuroprotective Agents; White Matter

Spinal cord injury (SCI) is a devastating disease inflicting lifetime disability to the victims. Military personnel are quite often victims of SCI for which no suitable therapeutic strategies have been developed so far. The main reason for SCI induced disability is loss of neural connections below and above the lesion site causing motor paralysis and somatosensory disturbances Loss of neuronal connections thwart spinal cord conduction resulting in motor function disability. To enhance spinal cord conduction grafting of peripheral nerves, implant of hydrogels filled with neuroprotective drugs is used but so far, no satisfactory results re achieved. In this regards implants of microelectrode for enhancing tissue connectivity is suggested that is still under experimental state. We have used titanium implant with or without TiO

Topics: Animals; Benzofurans; Capillary Permeability; Drug Delivery Systems; Drug Implants; Edema; Evoked Potentials; Locomotion; Male; Nanostructures; Nanowires; Neuroprotective Agents; Rats; Spinal Cord Injuries; Titanium

To study the effect of 3-n-butylphthalide (NBP) on neuronal apoptosis in rats with cerebral infarction (CI) through the p38/mitogen-activated protein kinase (MAPK) pathway.. A total of 30 rats were divided into control group (healthy rats, n=10), model group (CI rat model, n=10), and NBP group (CI rat model + intraperitoneal injection of NBP, n=10). Then, the neurological function, degree of cerebral ischemia, apoptosis of brain tissues, the protein and messenger ribonucleic acid (mRNA) expressions of p-p38 and MAPK in brain tissues were detected using the neurological score, 2,3,5-triphenyltetrazolium chloride (TTC) staining, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and Western blotting, respectively.. In NBP group, the neurological score was significantly lower than in model group, and the difference was statistically significant (p<0.05). The results of TTC staining revealed that the area of the white region in brain slices was significantly larger in model group than in control group, indicating the successful establishment of middle cerebral artery occlusion (MCAO) model. Compared with model group, NBP group had a smaller area and lighter color of the white region in brain slices, suggesting that NBP markedly reduces the MCAO-induced CI. The apoptosis rate in NBP group was higher than in control group (p<0.05), but lower than in model group (p<0.05), while it was higher in model group than in control group (p<0.05). The protein expressions of p38 and MAPK in NBP group were higher than in control group (p<0.05), but lower than in model group (p<0.05), while they were higher in model group than in control group (p<0.05). Moreover, the mRNA expressions of p38 and MAPK were lower in control group than in model group (p<0.05), while they were higher in model group than in NBP group (p<0.05), but there was no significant difference in the mRNA expression of p38 between NBP group and control group (p>0.05).. NBP alleviates neuronal apoptosis in CI by down-regulating the p38 signal and inhibiting the expression of MAPK, thereby treating CI.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Benzofurans; Cerebral Infarction; Disease Models, Animal; Down-Regulation; Female; Gene Expression Regulation; Injections, Intraperitoneal; MAP Kinase Signaling System; Neurons; Neuroprotective Agents; Proto-Oncogene Proteins c-bcl-2; Rats

Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB) is a novel pro-drug of 3-n-butylphthalide (dl-NBP) that is used to treat ischemic stroke. Currently, dl-PHPB is in phase II-III clinical trials in China. In this study, we investigated the conversion and pharmacokinetics profiles of dl-PHPB in vitro and in vivo. The conversion of dl-PHPB to dl-NBP was pH- and calcium-dependent, and paraoxonase was identified as a major enzyme for the conversion in rat plasma. The pharmacokinetics, tissue distribution and excretion of dl-PHPB were studied and compared with equal-molar doses of dl-NBP in rats and dogs. The in vivo studies showed that dl-PHPB could be quickly and completely converted to dl-NBP. The plasma concentration-time course of converted dl-NBP after intravenous dl-PHPB administration was nearly the same as that after equal-molar dl-NBP. The C

Topics: Adipose Tissue; Animals; Aryldialkylphosphatase; Benzoates; Benzofurans; Brain; Dogs; Feces; Female; Gastric Mucosa; Hydrogen-Ion Concentration; Male; Pentanes; Prodrugs; Rats, Sprague-Dawley

Oxidative stress induced by chronic cerebral hypoperfusion (CCH) plays an important role in the pathogenesis of vascular cognitive impairment (VCI). The Akt/Nrf2 signaling pathway is one of the most important antioxidative stress pathways. To explore whether NBP (DL-3-n-butylphthalide) could alleviate VCI induced by CCH via activating the Akt/Nrf2 signaling pathway and modifying the levels of apoptosis-related proteins, adult male Sprague-Dawley rats were subjected to permanent occlusion of bilateral common carotid arteries (BCCAO) and treated either with vehicle or NBP (applied in two doses, 40 mg/kg and 80 mg/kg) while sham operated animals were treated with vehicle. Treatments were administered daily for 28 days. The obtained results indicate that both administrated doses of NBP significantly ameliorated the spatial learning and memory impairments as indicated by the Morris water maze test while Hematoxylin-Eosin staining revealed that morphological defects in the CA1 area of hippocampus were improved. Moreover, NBP reversed the BCCAO-induced downregulation of investigated oxidative stress-related proteins (p-Akt, t-Nrf2, n-Nrf2 and HO-1) along with the upregulation of pro-apoptotic molecule, Bax and reduction of the expression of anti-apoptotic protein, Bcl-2. According to presented results, NBP may have a protective effect against cognitive and morphological impairments induced by CCH via activation of Akt/Nrf2 signaling pathway and inhibition of apoptotic cascade.

Topics: Animals; Benzofurans; Brain Ischemia; Cognitive Dysfunction; Disease Models, Animal; Hippocampus; Male; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidative Stress; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction

As microglial activation is a key factor in the pathogenesis of Parkinson's disease (PD), drugs that target this process may help to prevent or delay the development of PD. The present study investigated the effects of dl‑3‑n‑butylphthalide (NBP) on microglia in a lipopolysaccharide (LPS)-induced PD mouse model. The mice were randomly divided into a blank control group, LPS control group and NBP + LPS treatment group. Mice in the treatment group were given an intragastric infusion of 120 mg/kg NBP daily for 30 days during the establishment of the PD mouse model. At 4 and 28 weeks post‑treatment, the motor behaviours of the mice in each group were observed using the rotarod test and the open field test. In addition, immunohistochemical staining was performed to determine the levels of activated microglia, tumour necrosis factor‑α and α‑synuclein, and the number of tyrosine hydroxylase (TH)‑positive cells in the substantia nigra. NBP significantly improved dyskinesia, reduced microglial activation, decreased nuclear α‑synuclein deposition and increased the survival of TH‑positive cells in the substantia nigra of LPS‑induced PD model mice. These findings suggested that NBP may exert its therapeutic effect by reducing microglial activation in a mouse model of PD.

Topics: alpha-Synuclein; Animals; Benzofurans; Disease Models, Animal; Dyskinesias; Gene Expression Regulation; Immunohistochemistry; Lipopolysaccharides; Male; Maze Learning; Mice; Mice, Inbred C57BL; Microglia; Motor Activity; Neuroprotective Agents; Parkinson Disease, Secondary; Rotarod Performance Test; Substantia Nigra; Tumor Necrosis Factor-alpha; Tyrosine 3-Monooxygenase

Agents against atrial structural remodeling (ASR) are thought to block the occurrence of atrial fibrillation (AF). The aim of this study was to investigate the effects of DL-3-n-butylphthalide (NBP) on ASR and AF formation in rats with heart failure (HF) induced by myocardial infarction. The heart failure rats established 1 week after ligating left anterior descending coronary artery were randomly treated with vehicle (HF group, n = 24), or treated with DL-3-n-butylphthalide (100 mg/kg body weight) (NBP group, n = 26) for 4 weeks. Eighteen rats that underwent the same surgery but without ligating artery treated with vehicle were used as sham group (n = 18). Echocardiography, AF inducibility test, atrial fibrosis, gap junction, cytokine expression and serum antioxidant capacity analysis were detected at follow-up. Treatment of NBP for 4 weeks significantly improved cardiac function (P < 0.05), reduced AF inducibility and duration time (P < 0.05), and attenuated atrial fibrosis (P < 0.05). NBP also up-regulated protein expression of both overall Cx43 and phosphorylated Cx43 (P < 0.05) and improved the distribution of Cx43. Furthermore, NBP significantly inhibited the expression of TNF-α, NF-κB, and TGF-β1 and up-regulated Nrf2 and HO-1 protein expression with an increased serum T-AOC, CAT, and SOD activities and a reduced serum MDA. Collectively, NBP prevented ASR and AF in rats with HF by inhibiting atrial fibrosis, resynchronizing gap junction remodeling through inhibiting TNF-α/NF-κB/TGF-β1-related inflammatory reactions, and up-regulating Nrf2/HO-1-mediated antioxidant effects. Therefore, NBP may be a promising agent as upstream therapy for the prevention of AF.

Topics: Animals; Atrial Remodeling; Benzofurans; Cardiotonic Agents; Heart Atria; Heart Diseases; Heme Oxygenase (Decyclizing); Male; NF-E2-Related Factor 2; NF-kappa B; Rats, Sprague-Dawley; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Multiple sclerosis (MS) is a long-lasting autoimmune disease of the central nervous system. Currently, the etiology of MS is not known. Experimental autoimmune encephalomyelitis (EAE), has been recognized as the most widely used animal models to study the molecular mechanisms underlying MS and the efficacy of potential drugs for treatment of MS. In the present study, we found that Dl-3-n-butylphthalide (NBP), a neuroprotective drug in ischemic brain injury, prevented development of disease in experimental autoimmune encephalomyelitis (EAE) and significantly reduced inflammatory factors and necroptosis-associated genes, including PGAM5 in the spinal cord tissues. Similarly, silence of PGAM5 in spinal cord also ameliorated the disease severity in the mice with EAE. Moreover, re-expression of PGAM5 counteracted the protective effect of NBP on the pathogenesis of EAE. Importantly, we found that both NBP and silence of PGAM5 inhibited cellular necroptosis and inflammation in microglia induced by TNFα plus zVAD-fmk. Meanwhile, overexpression of PGAM5 reactivated cellular necroptosis and inflammation suppressed by NBP in vitro. Taken together, our findings provide evidence that NBP can attenuate the progression of EAE by suppressing PGAM5-induced necroptosis and inflammation in microglia and represents a new therapeutic strategy for treating autoimmune diseases.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Apoptosis Regulatory Proteins; Benzofurans; Dose-Response Relationship, Drug; Encephalomyelitis, Autoimmune, Experimental; Male; Mice; Mice, Inbred C57BL; Microglia; Multiple Sclerosis; Necrosis; Neuroprotective Agents; Phosphoprotein Phosphatases; Treatment Outcome

DL-3-n-butylphthalide (NBP) is used in the treatment of ischemic stroke. It was demonstrated NBP also has a cardioprotective effect in acute myocardial infarction (MI) model. However, the chronic effects of NBP on ventricular function, remodeling, and arrhythmias in post-MI stage are unknown. This study was to investigate the effect of NBP on reducing ventricular remodeling and arrhythmias in post-MI stage. Post-MI rats were randomly treated with 100 mg/kg NBP daily (n = 21) or vehicle (n = 21) for 5 weeks. Sham-operated rats were treated with the same dose vehicle (n = 18). Echocardiographic assessment, ventricular arrhythmias inducibility test, morphological and collagen analysis, immunohistochemistry, and western blot were studied. NBP significantly improved cardiac function, inhibited the severity and inducibility of ventricular arrhythmias, reduced cardiac index, fibrosis and hypertrophy, improved the protein expression and distribution of Cx43 gap junction, and upregulated PI3k/Akt/Nrf2 pathway and the downstream antioxidant response elements (ARE), including heme oxygenase-1, Glutathione, Cu-Zn superoxide dismutase, and Fe/Mn superoxide dismutase. These results suggest NBP improves LV function and reduces ventricular arrhythmias by mitigating LV fibrosis, hypertrophy, and Cx43 gap junction remodeling. PI3k/Akt/Nrf2/ARE signaling pathway may contribute to its anti-ventricular remodeling effects.

Topics: Animals; Arrhythmias, Cardiac; Benzofurans; Cardiotonic Agents; Connexin 43; Fibrosis; Heart Ventricles; Myocardial Infarction; NF-E2-Related Factor 2; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats, Sprague-Dawley; Ventricular Function; Ventricular Remodeling

The aim of this study was to explore whether Dl-3-n-butylphthalide (DBT) could protect blood-brain barrier (BBB) of mice with experimental cerebral infarction and the relevant mechanism.. Adult male CD-1 mice were selected as the study objects. The permanent middle cerebral artery occlusion (MCAO) model was prepared by Longa's modified suture-occluded method. The mice were randomly divided into 3 groups: the sham operation group (Sham group), the cerebral infarction model group (CI group) and the DBT (120 mg/kg) intervention group (DBT group). Neurologic function deficits were evaluated by Longa's modified scoring method after 24 h of permanent MCAO. The wet and dry weight method was used for measuring water content in brain tissues. 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining method was applied to determine the volume of cerebral infarction. Changes in the protein and messenger ribonucleic acid (mRNA) expression levels of matrix metallopeptidase 9 (MMP-9), claudin-5, vascular endothelial growth factor (VEGF), glial fibrillary acidic protein (GFAP), NF-E2 related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1) in ischemic brain tissues were detected using immunohistochemistry, Western blotting and quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Ultrastructure changes in BBBs were observed under an electron microscope.. DBT improved the neurologic function deficits of mice and reduced the infarction volume of mice with cerebral infarction. DBT alleviated edema and decreased the permeability of BBBs of mice with cerebral infarction. DBT down-regulated the expression of MMP-9 and up-regulated the expression of claudin-5 in brain tissues of mice with cerebral infarction. DBT increased the expressions of VEGF and GFAP. DBT improved the ultrastructure in capillary endothelial cells of BBBs and increased the expressions of Nrf-2 and HO-1.. DBT may protect BBB by activating the Nrf-2/HO-1 signaling pathway, thus achieving its protective effect on the brain.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Brain; Cerebral Infarction; Heme Oxygenase-1; Male; Membrane Proteins; Mice; Neuroprotective Agents; NF-E2-Related Factor 2; Rats, Sprague-Dawley; Signal Transduction

The limited degree of functional recovery is closely associated with the condition of the periinfarct cortex after ischemic stroke. The model of oxygen-glucose deprivation in cultured neurons could partly simulate this condition. Proper dendritic morphology is crucial for the correct wiring of neuronal function. Hence, the question of how to facilitate the plasticity of neural dendrites is of great significance. DL-3-n-butylphthalide is an efficient medication for ischemic stroke. In this study, in addition to having neuroprotective effects, DL-3-n-butylphthalide could increase the number of primary and secondary dendrites and of dendritic tips as confirmed by Sholl analysis. This study further demonstrated that DL-NBP inactivates PI3K/AKT signaling to positively regulate dendritic branching.

Topics: Animals; Apoptosis; Benzofurans; Cell Hypoxia; Cells, Cultured; Chromones; Dendrites; Glucose; Morpholines; Neurons; Neuroprotective Agents; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction

Alzheimer's disease (AD) is characterized by extracellular accumulation of β-amyloid peptides (Aβ) and intracellular neurofibrillary tangles, along with cognitive decline and neurodegeneration. The cognitive deficit is considered to be due to the dysfunction of hippocampal neurogenesis. Although L-3-n-butylphthalide (L-NBP) has been shown beneficial effects in multiple AD animal models, the underlying molecular mechanisms are still elusive. In this study, we investigated the effects of L-NBP on neurogenesis both in vitro and in vivo. L-NBP promoted proliferation and migration of neural stem cells and induced neuronal differentiation in vitro. In APP/PS1 mice, L-NBP induced neurogenesis in the dentate gyrus and improved cognitive functions. In addition, L-NBP significantly increased the expressions of BDNF and NGF, tyrosine phosphorylation of its cognate receptor, and phosphorylation of Akt as well as CREB at Ser133 in the hippocampus of APP/PS1 mice. These results indicated that L-NBP might stimulate the proliferation, migration, and differentiation of hippocampal neural stem cells and reversed cognitive deficits in APP/PS1 mice. BDNF/TrkB/CREB/Akt signaling pathway might be involved.

Topics: Alzheimer Disease; Amyloid beta-Protein Precursor; Animals; Benzofurans; Cell Differentiation; Cell Movement; Cell Proliferation; Embryo, Mammalian; Humans; Intercellular Signaling Peptides and Proteins; Male; Maze Learning; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Nerve Tissue Proteins; Neural Stem Cells; Neurogenesis; Presenilin-1

To explore the effects of N-butylphthalide on the expressions of ZO-1 and claudin-5 in blood-brain barrier (BBB) in rats with acute carbon monoxide (CO) poisoning.. A total of 144 adult healthy male Sprague-Dawley (SD) rats were randomly divided into normal control group, CO poisoning group, and NBP treatment group, with 48 rats in each group. The acute CO poisoning model was reproduced in hyperbaric oxygen chamber, and all model rats were given hyperbaric oxygen therapy once daily. The rats in the normal control group were free to breathe fresh air. The rats in NBP treatment group were administered orally NBP 60 mg/kg twice a day at 2 hours after poisoning until death. The rats in normal control group and CO poisoning group were treated with equal amount of pure olive oil. Four rats were sacrificed from each group at 1, 3, 7, 14 days after model reproducing, respectively. The changes in ultrastructure of BBB were observed under transmission electron microscope. The expressions of ZO-1 and claudin-5 proteins were determined by immunofluorescence staining and Western Blot. The localization of the two target proteins was observed by immunofluorescence double staining. The correlation between the two proteins was analyzed by linear regression.. The ultrastructure of BBB was normal in normal control group, some ZO-1 and a large number of claudin-5 positive cells were observed. The ultrastructure of BBB was seriously injured, ZO-1 and claudin-5 positive cells in brain tissue were significantly decreased, and the expressions of ZO-1 and claudin-5 proteins in brain tissue at 1 day after poisoning in CO poisoning group were significantly lower than those of normal control group (ZO-1 protein: 3.38±0.30 vs. 24.50±5.62, claudin-5 protein: 11.38±0.93 vs. 46.35±6.88, both P < 0.05), and although gradually restored, they were maintained at relatively lower levels until 14 days as compared with those in normal control group (ZO-1 protein: 10.35±0.80 vs. 24.63±3.57, claudin-5 protein: 32.35±3.11 vs. 46.43±7.20, both P < 0.05). NBP treatment could significantly alleviate the ultrastructure injury of BBB induced by acute CO poisoning, the amount of ZO-1 and claudin-5 positive cells in brain tissue were significantly increased, as well as the expressions of ZO-1 and claudin-5 proteins were significantly increased, which were significantly higher than those of CO poisoning group from 1 day and 3 days on, respectively (1-day ZO-1 protein: 7.57±0.69 vs. 3.38±0.30, 3-day claudin-5 protein: 20.46±1.42 vs. 11.43±0.86, both P < 0.05), and which showed an increase tendency with time prolongation. The results of immunofluorescence double staining showed that ZO-1 and claudin-5 proteins could not only coexist in the same cell, but also could be expressed separately in different cells. Linear regression analysis showed the positive correlation between the expressions of ZO-1 and claudin-5 proteins in brain tissue of rats with acute CO poisoning (R. NBP could markedly improve the ultrastructure and functional integrity of BBB through up-regulating the expressions of ZO-1 and claudin-5 proteins, and then reduce brain damage caused by CO poisoning.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Carbon Monoxide Poisoning; Claudin-5; Male; Rats; Rats, Sprague-Dawley

Dl-3-n-Butylphthalide (NBP), a small molecule compound extracted from the seeds of Apium graveolens, possesses a large range of biological effects. Here, we attempted to explore the therapeutic effects of NBP on lipopolysaccharide (LPS)-induced major depressive disorder (MDD) and gain further insight into the underlying mechanisms of the antidepressant effects of NBP.. We evaluated the effect of NBP against LPS-induced behavioral changes in rats. We also examined the inflammation, oxidative stress, and apoptosis markers and analyzed the Nrf2 and NF-κB pathways in the hippocampus of rats following repeated peripheral immune challenge by LPS for 2 weeks (500 μg/kg every other day).. Our results indicated that repeated LPS administration induced the rats to a depressive-like state and activated inflammatory response, oxidative stress, and apoptosis reactions in the hippocampus. NBP treatment attenuated the LPS-induced abnormal behavior and ameliorated pathogenic processes in rats with MDD. NBP reduced the inflammatory response with inhibited expression of pro-inflammatory cytokines including IL-1β and IL-6 and downregulated the NF-κB signal pathway. Concurrent with the anti-inflammation action, NBP reduced LPS-induced oxidative reactions in the hippocampus and enhanced Nrf2-targeted signals, as evidenced by increased transcription of antioxidant enzymes and decreased malondialdehyde (MDA) production. In addition, NBP inhibited LPS-induced neuronal apoptosis in the rat brain, as evidenced by decreased apoptosis marker Caspase-3 production and TUNEL assay.. These results provide more insight into pathogenesis of MDD and firstly demonstrated the potential antidepressant actions of NBP.

Topics: Animals; Antioxidants; Benzofurans; Cytokines; Depression; Lipopolysaccharides; Male; Neuroprotective Agents; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Rats; Rats, Sprague-Dawley; Signal Transduction

The aim of this study was to explore the role of DL-3-n-butylphthalide (NBP) in cerebral ischemia-reperfusion injury (CIRI) mice model. The involvement of extracellular signal-regulated kinase (ERK) signaling pathway was also investigated.. All mice were divided into five groups: sham-operated group, CIRI group, NBP pretreatment group, NBP treatment group, and NBP pretreatment + treatment group. The CIRI mice model was established by the use of the Pulsinelli four-vessel occlusion method. Pretreatment mice received NBP (90 mg/kg/d) three times a day within four days before reperfusion by gavage. Treatment mice received NBP (90 mg/kg/d) three times a day within five days after reperfusion by gavage. We detected the infarction area, the neurological severity, and the superoxide dismutase and malondialdehyde levels. Furthermore, we observed the expressions of GRASP65, phosphorylation of GRASP65 (pGRASP65), ERK, and phosphorylation of ERK (pERK) by the use of Western blotting.. The result showed that the ERK pathway was activated in response to CIRI. NBP decreases the expressions of pERK and pGRASP65 following CIRI. Additionally, NBP could decrease MDA and increase SOD level in brain tissues. Decreased infarct volume was also observed in the NBP group. Thereby, NBP inhibited the activation of the ERK pathway induced by CIRI and reduced the GRASP65 phosphorylation.. The current finding suggested that NBP protected the cerebrum from CIRI mediated by inhibiting the ERK signaling pathway and subsequently reducing GRASP65 phosphorylation.

Topics: Animals; Apoptosis; Benzofurans; Brain Ischemia; Carrier Proteins; Disease Models, Animal; Female; Intracellular Signaling Peptides and Proteins; Male; MAP Kinase Signaling System; Membrane Proteins; Mice; Mice, Inbred ICR; Neuroprotective Agents; Phosphorylation; Reperfusion Injury; Signal Transduction

L-3-n-butylphthalide (L-NBP) is a type of anti-ischemic cranial nerve protective drug that may act on vascular dementia (VD). Phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT/PKB) signaling pathway can up-regulate B-cell lymphoma 2 (Bcl-2) expression, reduce reactive oxygen species (ROS) production, and alleviate cell apoptosis. This study aimed at investigating the role of L-NBP on neurological function and cell apoptosis in VD mouse through regulating PI3K/AKT signaling pathway.. The mice were divided into four groups, including Sham, VD, VD + solvent, and VD + L-NBP. HT22 cells were cultured in vitro and treated by ischemia/reperfusion (I/R). HT22 cells were divided into four groups, including I/R, VD + solvent, VD + L-NBP, and VD + L-NBP + LY294002 groups. Phosphorylated AKT (p-AKT) and Bcl-2 expressions were tested. ROS content in hippocampus tissue was detected by flow cytometry. Cell apoptosis was evaluated by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay.. ROS content and cell apoptosis increased, while p-AKT and Bcl-2 expressions reduced in hippocampus tissue from VD group compared with sham group. L-NBP significantly up-regulated p-AKT and Bcl-2 expressions and decreased ROS content and cell apoptosis in hippocampus tissue. I/R treatment markedly induced HT22 cell apoptosis and ROS production, and reduced p-AKT and Bcl-2 expressions. L-NBP treatment markedly up-regulated p-AKT and Bcl-2 levels, restrained cell apoptosis, and reduced ROS content in TH22 cells intervened by I/R. LY294002 apparently attenuated the protective effect of L-NBP on HT22 cells.. L-NBP protects VD by up-regulating PI3K/AKT signaling pathway, elevating Bcl-2 expression, reducing nerve cell apoptosis, and restraining ROS production.

Topics: Animals; Apoptosis; Benzofurans; Dementia, Vascular; Hippocampus; Male; Mice; Mice, Inbred C57BL; Phosphatidylinositol 3-Kinase; Phosphorylation; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Reperfusion Injury; Signal Transduction

Doxorubicin (DOX) is a broad-spectrum antitumor drug while its use is limited due to its neurobiological side effects associated with depression. We investigated the neuroprotective efficacy of dl-3-n-butylphthalide (dl-NBP) against DOX-induced anxiety- and depression-like behaviors in rats. dl-NBP was given (30 mg/kg) daily by gavage over three weeks starting seven days before DOX administration. Elevated plus maze (EPM) test, forced swimming test (FST), and sucrose preference test (SPT) were performed to assess anxiety- and depression-like behaviors. Our study showed that the supplementation of dl-NBP significantly mitigated the behavioral changes induced by DOX. To further explore the mechanism of neuroprotection induced by dl-NBP, several biomarkers including oxidative stress markers, endoplasmic reticulum (ER) stress markers, and neuroinflammatory cytokines in the hippocampus were quantified. The results showed that dl-NBP treatment alleviated DOX-induced neural apoptosis. Meanwhile, DOX-induced oxidative stress and ER stress in the hippocampus were significantly ameliorated in dl-NBP pretreatment group. Our study found that dl-NBP alleviated the upregulation of malondialdehyde (MDA), nitric oxide (NO), CHOP, glucose-regulated protein 78 kD (GRP-78), and caspase-12 and increased the levels of reduced glutathione (GSH) and activities of catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPx) in the hippocampus of rats exposed to DOX. Additionally, the gene expression of interleukin-6 (IL-6), interleukin-1

Topics: Antibiotics, Antineoplastic; Behavioral Symptoms; Benzofurans; Doxorubicin; Endoplasmic Reticulum Stress; Humans; Inflammation; Neuroprotective Agents; Oxidative Stress

This study examined whether the neuroprotective drug, 3-. PC12 neuronal cells were pretreated for 24 hours with NBP (10 μmol/L), then exposed to oxygen and glucose deprivation (OGD) for 8 hours as an in vitro model of ischemic stroke. Indices of anti-oxidative response, mitochondrial function and mitochondrial dynamics were evaluated.. OGD suppressed cell viability, induced apoptosis and increased caspase-3 activity. NBP significantly reversed these effects. NBP prevented oxidative damage by increasing the activity of superoxide dismutase and lowering levels of malondialdehyde (MDA) and reactive oxygen species (ROS). At the same time, it increased expression of Nrf2, HO-1 and AMPK. NBP attenuated mitochondrial dysfunction by enhancing mitochondrial membrane potential and increasing the activity of mitochondrial respiratory chain complexes I-IV and ATPase. NBP altered the balance of proteins regulating mitochondrial fusion and division.. NBP exerts neuroprotective actions by enhancing anti-oxidation and attenuating mitochondrial dysfunction. Our findings provide insight into how NBP may exert neuroprotective effects in ischemic stroke and raise the possibility that it may function similarly against other neurodegenerative diseases involving mitochondrial dysfunction.

Topics: Animals; Antioxidants; Apoptosis; Benzofurans; Brain Ischemia; Membrane Potential, Mitochondrial; Mitochondria; Mitochondrial Dynamics; Neurons; Neuroprotective Agents; Oxidative Stress; PC12 Cells; Rats; Signal Transduction; Stroke

To evaluate the neuroprotective effects of Dl-3-n-butylphthalide (NBP) on patients with radiation-induced brain injury, a hospital-based, clinical retrospective cohort study was conducted.. Data were collected on patients diagnosed with radiation-induced brain injury from January 2009 to January 2015 in Department of Neurology, Sun Yat-Sen Memorial Hospital. All patients enrolled have received cranial radiotherapy and were diagnosed with radiation-induced brain injury. Patients fulfilling certain eligibility criteria were recruited for analysis. The clinical therapeutic effects were observed and evaluated by LENT/SOMA scores before and one month after treatment in these two groups, respectively.. The therapeutic effects of headache (total efficiency 75.76%), eurologic deficit (total efficiency 81.58%), cognitive functions (total efficiency 77.78%) and MRI results (total efficiency 74.29%) were better in the experimental group than those in the control group (p < 0.05). Nevertheless, there was no significant difference in mood and personality changes between these two groups.. Administration of Dl-3-n-butylphthalide, in adjunct to corticosteroid therapy, might provide a better outcome in patients with radiation-induced brain injury.

Topics: Adrenal Cortex Hormones; Adult; Benzofurans; Brain Injuries; Cranial Irradiation; Drug Therapy, Combination; Female; Humans; Male; Middle Aged; Neuroprotective Agents; Outcome Assessment, Health Care; Radiation Injuries; Retrospective Studies

Previous investigations suggest that DL-3-n-butylphthalide (NBP) is a promising multifaceted drug for the treatment of stroke. It is not clear whether NBP can treat traumatic brain injury (TBI) and what could be the mechanisms of therapeutic benefits. To address these issues, TBI was induced by a controlled cortical impact in adult male mice. NBP (100 mg/kg) or saline was intraperitoneally administered within 5 min after TBI. One day after TBI, apoptotic events including caspase-3/9 activation, cytochrome c release from the mitochondria, and apoptosis-inducing factor (AIF) translocation into the nucleus in the pericontusion region were attenuated in NBP-treated mice compared to TBI-saline controls. In the assessment of the nuclear factor kappa-light-chain-enhancer of activated B (NF-κB) pathway, NBP ameliorated the p65 expression and the p-IκB-α/IκB-α ratio, indicating reduced NF-κB activation. Consistently, NBP reduced the upregulation of proinflammatory cytokines such as tumor necrotizing factor-alpha (TNF-α) and interleukin-1beta (IL-1β) after TBI. In sub-acute treatment experiments, NBP was intranasally delivered once daily for 3 days. At 3 days after TBI, this repeated NBP treatment significantly reduced the contusion volume and cell death in the pericontusion region. In chronic experiments up to 21 days after TBI, continues daily intranasal NBP treatment increased neurogenesis, angiogenesis, and arteriogenesis in the post-TBI brain, accompanied with upregulations of regenerative genes including brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), endothelial-derived nitric oxide synthase (eNOS), and matrix metallopeptidase 9 (MMP-9). The NBP treatment significantly improved sensorimotor functional recovery and reduced post-TBP depressive behavior. These new findings demonstrate that NBP shows multiple therapeutic benefits after TBI.

Topics: Animals; Behavior, Animal; Benzofurans; Brain Injuries, Traumatic; Cytokines; Disease Models, Animal; Interleukin-1beta; Male; Mice, Inbred C57BL; Neuroprotection; NF-kappa B; Recovery of Function; Signal Transduction; Up-Regulation

Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB) is a prodrug of 3-n-butylphthalide (dl-NBP) for treatment of cerebral ischemic stroke in China, which undergoes lactonization to form dl-NBP in plasma. And, the phase II-III clinical trial of dl-PHPB has been approved by China Food and Drug Administration (CFDA) in 2013. In this study, a toxicity and toxicokinetics evaluation of dl-PHPB was performed using beagle dogs at specially high-dose 108 mg/kg/day (65-fold higher than humans at MHRD) for 4 weeks by intravenous administration, with a 3-week recovery period. And the plasma concentrations of dl-PHPB along with its metabolite dl-NBP were determined by HPLC-UV method. The results showed that dl-PHPB was quickly metabolized into dl-NBP, and no significant accumulation was observed. A slight to moderate behavior-associated toxicity was revealed in the process of delivery; and recovered to normal at the end of administration. Changes in the blood hematological profiles included significantly increased NEUT levels and lower LYM% content. Meanwhile, a notable increase in TG content was also observed in the serum biochemical parameters at 4-week post-exposure. These findings were reversible during the recovery period. The information from these studies would be taken into consideration for the interpretation of toxicology findings and provide a reference for clinical safety assessment.

Topics: Animals; Benzoates; Benzofurans; China; Dogs; Female; Humans; Male; Molecular Structure; Pentanes; Potassium; Toxicokinetics

Racemic l-3-n-butylphthalide (dl-NBP), is able to achieve a functional recovery in animal models of cerebral ischemia, vascular dementia, and Alzheimer's disease. In this study, we investigated the effect of dl-NBP on axonal growth, neurogenesis and behavioral performances in rats with cerebral ischemia.. Focal cerebral ischemia in rats was produced by intracerebral injection of endothelin-1. Starting from postoperative day 7, the experimental rats were administered 70mg/kg dl-NBP by oral gavage for two weeks. Biotinylated dextran amine (BDA) was injected into the contralateral sensorimotor cortex on day 14 after ischemia to trace the sprouting of corticospinal tract (CST) fibers into the denervated cervical spinal cord. The expressions of Nogo-A, Nogo-R, Rho-A, and ROCK in the perilesional cortex, the expressions of BDA, PSD-95, and vGlut1 in the denervated spinal cord, 5-bromo-20-deoxyuridine (BrdU)/DCX-positive cells in the subventricular zone (SVZ) of the injured hemisphere were detected by immunofluorescence. The rats' behavioral abilities were measured on postoperative days 30-32 in the beam-walking, cylinder and sticky label tests.. dl-NBP treatment significantly increased the number and length of crossing CST fibers, enhanced significantly the expression levels of synapse-associated proteins including PSD95 and VGlut-1 in the denervated cervical spinal cord, elevated the number of BrdU+/DCX+ cells in SVZ, and reduced markedly those of Rho-A+, ROCK+, Nogo-A+ and Nogo-R+ cells in perilesional cortex. In addition, dl-NBP improved the behavioral performance of the ischemic rats.. dl-NBP enhanced the behavioral recovery after cerebral ischemia in rats, possibly by increasing axonal growth and neurogenesis.

Topics: Animals; Benzofurans; Biotin; Brain Infarction; Dextrans; Disease Models, Animal; Disks Large Homolog 4 Protein; Doublecortin Protein; Endothelin-1; Male; Motor Activity; Neuronal Plasticity; Nogo Proteins; Nogo Receptor 1; Platelet Aggregation Inhibitors; Psychomotor Performance; Rats; Rats, Wistar; Recovery of Function; Signal Transduction; Stroke; Vesicular Glutamate Transport Protein 1

Myocardial infarction (MI) is an acute and fatal condition that threatens human health. Dl-3-n-butylphthalide (NBP) has been used for the treatment of acute ischemic stroke. Mitochondria may play a protective role in MI injury. However, there are few reports on the cardioprotective effect of NBP or the potential mitochondrial mechanism for the NBP-induced protection against cardiac ischemia injury. We investigated the therapeutic effects of NBP in an in vivo MI model and an in vitro oxidative stress model, as well as the potential mitochondrial mechanism.. This study comprised two different experiments. The aim of experiment 1 was to determine the protective effects of NBP on MI and the underlying mechanisms in vivo. In part 1, myocardial infarct size was measured by staining with 2,3,5-triphenyltetrazoliumchloride (TTC). Myocardial enzymes and mitochondrial enzymes were assayed. The aim of experiment 2 was to investigate the role of NBP in H. NBP treatment significantly reduced the infarct ratio, as observed by TTC staining, decreased serum myocardial enzymes in MI, and restored heart mitochondrial enzymes (isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), and a-ketoglutarate dehydrogenase (a-KGDH) activities after MI. Moreover, in in vitro studies, NBP significantly increased the viability of H9c2 cells in a dose-dependent manner, reduced cell apoptosis, protected mitochondrial functions, elevated the cellular ATP levels, and promoted H. Collectively, the results from both the in vivo and in vitro experiments suggested that NBP exerted a cardioprotective effect on cardiac ischemic injury via the regulation of mitochondrial function and biogenesis.

Topics: Animals; Benzofurans; Cardiotonic Agents; Cell Line; Cell Survival; Dose-Response Relationship, Drug; Heart; Membrane Potential, Mitochondrial; Mitochondria; Myoblasts, Cardiac; Myocardial Infarction; Neuroprotective Agents; Organelle Biogenesis; Oxidative Stress; Rats

L-3-n-Butylphthalide (L-NBP) exerts neuroprotective effects in animal models of cerebral ischemia, but its potential benefits in repeated cerebral ischemia-reperfusion (RCIR) injury remain unknown. We investigated the effect of L-NBP on cognitive impairment induced by RCIR in mice. Male C57Bl/6 mice received sham surgery or bilateral common carotid artery occlusion (3 times, 20 min each) and were orally administered preoperative L-NBP (30 mg/kg/day, 7 days), postoperative L-NBP (30 or 60 mg/kg/day, 28 days) or postoperative vehicle (28 days). Learning and memory were assessed by the Morris water maze task and step-down passive avoidance test. Nissl staining was used to identify pathologic changes in the hippocampal CA1 region. The expressions of proteins associated with signaling, apoptosis and autophagy were assessed by quantitative PCR and western blot. RCIR induced deficits in learning and memory that were alleviated by preoperative or postoperative L-NBP administration. Pathologic lesions in the hippocampal CA1 region induced by RCIR were less severe in mice treated with L-NBP. Preoperative or postoperative L-NBP administration in mice receiving RCIR promoted hippocampal expression of phospho-Akt and phospho-mTOR (suggesting activation of Akt/mTOR signaling), increased the Bcl-2/Bax ratio (indicating suppression of apoptosis) and reduced the LC3-II/LC3-I ratio (implying inhibition of autophagy). Preoperative or postoperative L-NBP administration also depressed hippocampal levels of beclin-1 mRNA (indicating suppression of autophagy). These findings suggest that the effect of L-NBP to alleviate learning and memory deficits in mice following RCIR may involve activation of Akt/mTOR signaling and regulation of the expressions of proteins related to apoptosis and autophagy.

Topics: Animals; Apoptosis; Autophagy; Benzofurans; Brain Ischemia; Cognition Disorders; Cognitive Dysfunction; Disease Models, Animal; Male; Memory Disorders; Mice, Inbred C57BL; Neuroprotective Agents; Proto-Oncogene Proteins c-akt; Reperfusion; TOR Serine-Threonine Kinases

In this study, we examined the neuroprotective effects and anti-inflammatory properties of Dl-3-n-butylphthalide (NBP) in Sprague-Dawley (SD) rats following traumatic spinal cord injury (SCI) as well as microglia activation and inflammatory response both in vivo and in vitro. Our results showed that NBP improved the locomotor recovery of SD rats after SCI an significantly diminished the lesion cavity area of the spinal cord, apoptotic activity in neurons, and the number of TUNEL-positive cells at 7 days post-injury. NBP inhibited activation of microglia, diminished the release of inflammatory mediators, and reduced the upregulation of microglial TLR4/NF-κB expression at 1 day post-injury. In a co-culture system with BV-2 cells and PC12 cells, NBP significantly reduced the cytotoxicity of BV-2 cells following lipopolysaccharide (LPS) stimulation. In addition, NBP reduced the activation of BV-2 cells, diminished the release of inflammatory mediators, and inhibited microglial TLR4/NF-κB expression in BV-2 cells. Our findings demonstrate that NBP may have neuroprotective and anti-inflammatory properties in the treatment of SCI by inhibiting the activation of microglia via TLR4/NF-κB signalling.

Topics: Animals; Anti-Inflammatory Agents; Benzofurans; Cell Line; Coculture Techniques; Female; Gene Expression Regulation; Lipopolysaccharides; Microglia; Neuroprotective Agents; NF-kappa B; PC12 Cells; Rats; Rats, Sprague-Dawley; Signal Transduction; Spinal Cord; Spinal Cord Injuries; Toll-Like Receptor 4

DL‑3‑n‑butylphthalide (NBP) is extracted from rapeseed and exhibits multiple neuroprotective effects, exerted by inhibiting the inflammatory process, including reducing oxidative stress, improving mitochondrial function and reducing neuronal apoptosis. The present study aimed to investigate the neuroprotective effects of NBP in a lipopolysaccharide (LPS)‑induced mouse model of Parkinson's disease (PD). The behavior of mice was assessed using the rotarod test and open‑field test, the amount of tyrosine hydroxylase in the substantia nigra pars compacta was evaluated by immunohistochemistry, and the levels of phosphorylated c‑Jun N‑terminal kinase (JNK), mitogen‑activated protein kinase 14 (p38) and extracellular signal‑regulated kinase 1 were determined by western blotting. It was demonstrated that the LPS‑induced behavioral deficits were significantly improved. LPS‑induced dopaminergic neurodegeneration was relieved following treatment with NBP, as determined from tyrosine hydroxylase‑positive cells. Phosphorylation of JNK and p38 was significantly inhibited following treatment with NBP. Therefore in the present study, a role for NBP has been established in the treatment of a PD murine model, laying the experimental basis for the treatment of PD with this agent.

Topics: Animals; Apoptosis; Benzofurans; Disease Models, Animal; Dopamine; Dopaminergic Neurons; Lipopolysaccharides; Male; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Neuroprotective Agents; Oxidative Stress; Parkinson Disease; Rotarod Performance Test; Substantia Nigra; Tyrosine 3-Monooxygenase

The pathogenesis of vascular dementia (VD) is associated with neuronal degeneration, apoptosis or necrosis following ischemic brain injury. L‑butylphthalide (L‑NBP), has been demonstrated to exhibit potent anti‑ischemic and anti‑VD effects, however the associated specific mechanism remains to be elucidated. The present study generated a VD rat model, in which the effect of L‑NBP on neurological function and expression levels of brain‑derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB) were observed. A total of 90 male Sprague Dawley rats were randomly divided into sham, model and L‑NBP groups (n=30). The VD model was generated by ligation of bilateral common carotid artery. A Morris water maze was used to test learning and memory functions. Animals were then sacrificed and cortical and hippocampal tissues were extracted. Hematoxylin and Eosin staining was used to observe brain tissue injury, and reverse transcription‑quantitative polymerase chain reaction was employed to measure BDNF and TrkB mRNA levels. Western blotting was employed to measure BDNF, TrkB and serine‑threonine protein kinase (Akt) protein levels. Immunohistochemistry staining was used to detect the N‑methyl‑D‑aspartate receptor (NMDAR) levels. VD rats exhibited elongated escape latency and lower crossing times, with significant neuronal damage. L‑NBP treatment shortened escape latency, increased crossing times and improved cortical and hippocampal injury. BDNF, TrkB, Akt and NMDAR expressions in the treatment group were significantly increased compared with the model group (P<0.05). L‑NBP may therefore enhance hippocampal expression of BDNF, TrkB, Akt and NMDAR, decrease ischemic injury of VD rats, and improve learning and memory.

Topics: Animals; Benzofurans; Brain-Derived Neurotrophic Factor; Carotid Artery, Common; Cerebral Cortex; Dementia, Vascular; Disease Models, Animal; Gene Expression Regulation; Hippocampus; Ligation; Male; Maze Learning; Memory, Short-Term; Neuroprotective Agents; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Receptor, trkB; Receptors, N-Methyl-D-Aspartate; Signal Transduction

Butylphthalide, a component extracted from seeds of Chinese celery, is an effective neuroprotective agent used for the treatment of ischemic stroke and dementia. Diabetes may cause central nervous system damage, and diabetes is closely associated with dementia. The aim of the present study was to investigate the effects of butylphthalide on cognitive impairment in a streptozotocin‑induced diabetic rat model, and the underlying mechanisms of action. A total of 30 healthy male Sprague Dawley rats were randomly divided into the following 2 groups: Normal control (NC; n=10) and diabetes model (DM) groups (n=20). Diabetes was induced in rats in the DM group by intraperitoneal injection of streptozotocin, and these rats were further subdivided into the following 2 groups: Diabetic control (n=10) and butylphthalide‑treated groups (n=10). Following 8 consecutive weeks of treatment, a Morris water maze test was performed and the levels of blood fasting plasma glucose (FPG), superoxide dismutase (SOD), malondialdehyde (MDA) and tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑1β, and IL‑6 inflammatory cytokines in the hippocampus were measured. FPG levels were significantly decreased in the butylphthalide‑treated group when compared with the DM group. In addition, cognitive deficits in diabetic rats were improved following butylphthalide treatment. Furthermore, butylphthalide significantly increased the level of SOD, reduced MDA levels, and reduced TNF‑α, IL‑1β, and IL‑6 levels in the hippocampus when compared with the DM group. The results of the present study suggest that butylphthalide may be an effective neuroprotective agent to improve cognitive dysfunction during diabetes.

Topics: Animals; Benzofurans; Blood Glucose; Body Weight; Brain-Derived Neurotrophic Factor; Cognition; Cognitive Dysfunction; Cytokines; Diabetes Complications; Diabetes Mellitus, Experimental; Disease Models, Animal; Fasting; Gene Expression Regulation; Hippocampus; Inflammation Mediators; Maze Learning; Neuroprotective Agents; Rats

Topics: Adenosine Triphosphate; Animals; Benzofurans; Brain Ischemia; Citric Acid; Disease Models, Animal; Glucose; Infarction; Metabolic Networks and Pathways; Neuroprotective Agents; Platelet Aggregation Inhibitors; Rats; Severity of Illness Index; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

To investigate the potential effect of 3-n-butylphthalide (NBP) pretreatment on the cerebral ischemia/reperfusion injury in rats and the relevant mechanism.. A total of 90 rats was divided into three groups: Sham operation group (Sham group), ischemia-reperfusion group (I-R group), and NBP pretreatment group (NBP group 75 mg·kg-1·d-1 gavage). Pre-treatment was given once a day within 1 week before establishing the rat model of cerebral ischemia-reperfusion injury. The middle cerebral artery occlusion (MACO) rat models were established with the improved Longa-Zea method on the 7th day after ischemia for 2 h and reperfusion for 24 h in all the rats. We detected the cerebral infarction, the pathologic change of brain, the apoptosis of nerve cell, the production levels of reactive oxygen species (ROS), the content of malonaldehyde (MDA) and the activity of superoxide dismutase (SOD), the water content and the permeability of blood-brain barriers (BBB). In addition, we also observed the expressions of mitogen-activated protein kinase (MAPK, p-38, JNK, ERK1/2) and cleaved caspase-3 in the hippocampus tissues.. Compared with Sham group, we discovered that NBP significantly reduced infarction area, cell apoptosis, BBB damage and water content. Further, we found that NBP could also decrease ROS and MDA, and increase SOD activity in brain tissues of rats with a cerebral ischemia-reperfusion injury. Moreover, results showed that NBP also inhibited the levels p38 and JNK.. NBP protected the cerebral from I/R injury, providing ideas for the expansion of clinical adaptability of NBP and possible approaches for its application.

Topics: Animals; Apoptosis; Benzofurans; Blood-Brain Barrier; Body Water; Brain Ischemia; Caspase 3; Hippocampus; Infarction, Middle Cerebral Artery; Malondialdehyde; MAP Kinase Signaling System; Neurons; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion Injury; Superoxide Dismutase

After spinal cord injury (SCI), the destruction of blood-spinal cord barrier (BSCB) is shown to accelerate gathering of noxious blood-derived components in the nervous system, leading to secondary neurodegenerative damages. SCI activates endoplasmic reticulum stress (ER stress), which is considered to evoke secondary damages of neurons and glia. Recent evidence indicates that Dl-3-n-butylphthalide (NBP) has the neuroprotective effect in ischaemic brain injury, but whether it has protective effects on SCI or not is largely unclear. Here, we show that NBP prevented BSCB disruption after SCI via inhibition of ER stress. Following a moderate contusion injury of the T9 level of spinal cord, NBP was administered by oral gavage and further treated once a day. NBP significantly attenuated BSCB permeability and breakdown of adherens junction (AJ) and tight junction (TJ) proteins, then improved locomotion recovery following SCI. The protective role of NBP on BSCB disruption is associated with the restrain of ER stress caused by SCI. Furthermore, NBP considerably constrained the expression of ER stress-associated proteins and degradation of TJ and AJ in human brain microvascular endothelial cells (HBMECs) treated with TG. In conclusion, our results indicate that ER stress is associated with the disruption of BSCB integrity after injury, NBP attenuates BSCB disruption via inhibiting ER stress and improve functional recovery following SCI.

Topics: Adherens Junctions; Animals; Benzofurans; Blood-Brain Barrier; Cells, Cultured; Endoplasmic Reticulum Stress; Female; Humans; Locomotion; Rats; Rats, Sprague-Dawley; Recovery of Function; Spinal Cord; Spinal Cord Injuries; Tight Junctions

To study the anti-asthmatic effects of butylphthalide in guinea pig.. Butylphthalide at the concentrations of 1、10、100 mg/L had an-ti-spasmodic effects on spasmodical tracheal smooth muscle of guinea pig (15.08 ±7.68、42.41 ±13.54、77.56 ±24.82 to acetylcholine, 19.40 ±7.60、56.84 ±11.72、76.35 ±19.40 to histamine), which showed a certain dose-effect relationship. Butylphthalide could prolong asth-matic incubation period (53.3 ±13.2、33.1 ±13.0), improve asthmatic behaviors, reduce NO in serum (78.71 ±19.40、84.75 ±20.97) and ET-1 in bronchoalveolar lavage fluid (24.30 ±5.80、28.50 ±6.31) (. Butylphthalide has some effects of anti-asthma and one of the mechanisms is to relieve abnormal increase of NO and ET-1.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Benzofurans; Endothelin-1; Guinea Pigs; Muscle, Smooth; Nitric Oxide; Trachea

Considering the complex etiology of Alzheimer's disease (AD), multifunctional agents may be beneficial for the treatment of this disease. A series of DL-3-n-butylphthalide-Edaravone hybrids were designed, synthesized and evaluated as novel dual inhibitors of amyloid-β aggregation and monoamine oxidases. Among them, compounds 9a-d exhibited good inhibition of self-induced Aβ

Topics: Alzheimer Disease; Amyloid beta-Peptides; Antioxidants; Antipyrine; Benzofurans; Binding Sites; Blood-Brain Barrier; Edaravone; Humans; Hydrogen Bonding; Inhibitory Concentration 50; Molecular Docking Simulation; Monoamine Oxidase; Monoamine Oxidase Inhibitors; Protein Binding; Protein Structure, Tertiary

The present study investigated the effects of dl-3-n-butylphthalide on cognitive function of patients with acute ischemic stroke (AIS).. A total of 104 patients with AIS admitted between October 2012 and June 2013 were assigned to either the Treatment (standardized treatment plus dl-3-n-butylphthalide) or Control (standardized treatment alone) groups. Cognitive function was assessed by the Beijing version of the Montreal Cognitive Assessment (MoCA-BJ) and Mini-Mental State Examination (MMSE) before and 1 month after treatment, when high-sensitivity C-reactive protein (hs-CRP) and homocysteine (Hcy) were also detected. A multivariate logistic regression analysis was done for explore the independent risk factors for vascular dementia (VD).. The proportion of cognitive impairment was significantly lower after treatment than before in both the Treatment (88% vs. 64%, P = 0.023) and Control (87% vs. 70%, P = 0.047) groups. Vascular dementia dropped from 30 to 10% in the Treatment (P = 0.035) and from 25.9 to 16.7% in the Control (P = 0.027) groups. Total cognitive improvement was more significant in the Treatment Group (P = 0.018); naming, memory, attention, and linguistic abilities were significantly improved (all P < 0.05). Serum Hcy and hs-CRP levels were significantly lower in the Treatment Group than in the Control Group 1 month after treatment (P < 0.05).. Dl-3-n-butylphthalide could significantly improve the cognitive function of AIS patients 1 month after stroke. Hcy was involved in the incidence of VD 1 month after AIS. However, further studies are necessary because of differences between groups at baseline.

Topics: Benzofurans; Biomarkers; Brain Ischemia; C-Reactive Protein; Cognition; Dementia, Vascular; Female; Homocysteine; Humans; Logistic Models; Male; Mental Status Schedule; Middle Aged; Multivariate Analysis; Neuroprotective Agents; Neuropsychological Tests; Nootropic Agents; Prospective Studies; Risk Factors; Stroke; Treatment Outcome

The present study was aimed to evaluate the neuroprotective effects of NBP in the mice models of TBI, as well as the possible role of Nrf2-ARE pathways in the assumptive neuroprotection. In mice,a modified Marmarou's weight-drop model was employed to induce TBI. ICR mice were randomly assigned to four experimental groups: sham, TBI, TBI+vehicle(V) and TBI+NBP. NBP (100 mg/kg) was administered via an intraperitoneal (i.p.) injection at 1 h following TBI. The administration of NBP significantly ameliorated the effects of the brain injury, including neurological deficits, brain water content, and cortical neuronal apoptosis. Furthermore, the level of malondialdehyde and the activity of superoxide dismutase (SOD) paired with glutathione peroxidase (GPx) were restored in the NBP treatment group. NBP promoted the translocation of Nrf2 protein from the cytoplasm to the nucleus markedly, increased the expressions of Nrf2-ARE pathway-related downstream factors, including hemeoxygenase-1(HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1), and prevented the decline of antioxidant enzyme activities, including SOD and GPx. NBP enhanced the translocation of Nrf2 to the nucleus from the cytoplasm,verified by a western blot, immunofluorescence. Additionally, it upregulated the expression of the Nrf2 downstream factors such as HO-1 and NQO1 were also confirmed via a western blot and real-time quantitative polymerase chain reaction. In conclusion, NBP administration may increase the activities of antioxidant enzymes and attenuate brain injury in a TBI model, potentially via the mediation of the Nrf2-ARE pathway.

Topics: Animals; Benzofurans; Brain Injuries, Traumatic; Disease Models, Animal; Male; Mice; Mice, Inbred ICR; Neuroprotective Agents; NF-E2-Related Factor 2; Oxidative Stress; Random Allocation; Signal Transduction

To study the effects of butylphthalide on bronchial asthma in guinea pigs, and investigate the involvement of endothelin.. In guinea pigs, bronchial asthma was induced by injection of ovalbumin (OVA) and provoked by inhalation of OVA, and the effects of butylphthalide on asthma were evaluated through the changes it induced by OVA, pulmonary function, endothelin-1 (ET-1) contents and activity of endothelin converting enzyme-1 (ECE-1) in bronchoalveolar lavage fluid (BALF), serum and lung tissue, and the gene expression of ET-1 in lung tissue.. Butylphthalide significantly improved pulmonary function, lowered asthmatic behavior score, inhibited the activity of ECE-1, and reduced ET-1 gene expression level in lung tissue.. Butylphthalide has an anti-asthma effect and the mechanisms involve inhibition of ECE-1 activity and lowering of ET-1geng expression.

Topics: Animals; Asthma; Benzofurans; Bronchoalveolar Lavage Fluid; Endothelin-1; Guinea Pigs; Ovalbumin

Microglia activation-induced neuroinflammation contributes to neuronal damage in neurodegenerative diseases. Inhibition of microglia activation and reduction of major neurotoxic cytokines have been becoming a therapeutic strategy for neurodegenerative diseases. L-3-n-Butylphthalide (L-NBP) has shown the potent neuroprotective effects in stroke and Alzheimer's disease animal models. The present study investigated the immune modulatory effects of L-NBP on pro-inflammatory cytokines and microglia activation in brain tissue induced by systemic lipopolysaccharide (LPS) treatment in C57BL/6 mice. Our results showed that systemic LPS treatment induced microglia activation in the brain. L-NBP treatment significantly suppressed the expression of proinflammatory cytokines, such as tumor necrosis factor (TNFα), interlukin-1β (IL-1β), interlukin-6 (IL-6), and interlukin-10 (IL-10) in LPS-treated mice. At the meantime, L-NBP treatment decreased the morphological activation of microglia. In addition, the phosphorylation level of JNK MAP kinase-signaling pathway was also inhibited by L-NBP in LPS-treated mice. Furthermore, L-NBP upregulated the expression of heme oxygenase (HO)-1, a key element in the anti-inflammation and anti-oxidative stress. These results suggested that L-NBP might be a promising candidate in delaying and reversing the progress of neurodegenerative diseases by inhibiting microglia activation.

Topics: Animals; Benzofurans; Brain; Cytokines; Disease Models, Animal; Heme Oxygenase-1; Immunologic Factors; Inflammation; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Microglia; Mitogen-Activated Protein Kinases; Molecular Structure; Neuroprotective Agents; Phosphorylation; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

DL-3-n-butylphthalide (NBP) is a therapeutic drug used for ischemic stroke treatment. Here, we investigated the impact of NBP on the development of rat diabetic cataract induced by intraperitoneal injection of streptozotocin (STZ). NBP was then administrated by oral gavage for nine weeks. Cataract development was monitored through ophthalmoscope inspections. The levels of blood glucose and serum reactive oxygen species (ROS), malondialdehyde (MDA) and 8-Hydroxydeovexyguanosine (8-OHdG) were measured. Total and soluble protein and oxidative stress parameters, such as 2, 4- dinitrophenylhydrazone (DNP), 4-hydroxynonenal (4-HNE) and MDA in the lenses were determined by Western blot and thiobarbituric acid analyses. The expressions of NF-E2-related factor 2 (Nrf2) and its downstream antioxidant enzymes, thioredoxin (TRX), Catalase and nuclear accumulation of Nrf2 were determined by Western blot and immunohistochemistry analyses. We showed that NBP treatment significantly improved the cataract scores, the levels of DNP, 4-HNE, and MDA in the lens compared to the non-treated groups. NBP also enhanced the expressions of Nrf2, TRX and catalase in the lens of diabetic rats. In addition, NBP treatment also decreased levels of blood glucose, serum MDA and 8-OHdG. These results suggested that NBP treatment significantly delayed the onset and progression of diabetic cataract by inhibiting the oxidative stresses.

Topics: Animals; Benzofurans; Biomarkers; Blood Glucose; Body Weight; Catalase; Cataract; Diabetes Complications; Diabetes Mellitus, Experimental; Disease Models, Animal; Disease Progression; Immunohistochemistry; Lens, Crystalline; Male; NF-E2-Related Factor 2; Oxidative Stress; Rats; Thioredoxins

This study investigates the impacts of n-butylphthalide (NBP) on the expression of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) in rats with focal cerebral ischemia. The thread embolization method was used to prepare the rat model of cerebral ischemia-reperfusion (CIR). The animals were divided into a sham operation group, a model control group and NBP treatment group. The NBP group was orally administered 25 mg/kg NBP twice a day after the surgery. The immunohistochemistry and reverse transcription-polymerase chain reaction were performed to observe the protein and mRNA expressions of VEGF and TGF-β 16 hours, 1 day and 2 days after inducing CIR. The mRNA and protein expressions of VEGF and TGF-β1 in the model control group and the NBP treatment group were all increased after CIR, and those of the NBP treatment group at each post-CIR time point were higher than the model control group (p < 0.01). After CIR, the expressions of VEGF and TGF-β1 increased, suggesting that VEGF and TGF-β1 exhibited protective effects towards the ischemic brain injuries, and that NBP could upregulate the expressions of VEGF and TGF-β1 in the peri-infarcted area, thus possibly protecting the ischemic brain tissues through this mechanism.

Topics: Animals; Behavior, Animal; Benzofurans; Brain Ischemia; Cerebral Infarction; Intercellular Signaling Peptides and Proteins; Male; Neurologic Examination; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Reperfusion Injury; RNA, Messenger; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

The research aim was to investigate the effects of dl-3-n-butylphthalide (NBP) on the level of circulating endothelial progenitor cells (EPCs) and clinical outcome in patients with acute ischemic stroke (AIS).. A total of 170 patients were included and randomly assigned to NBP group and control group. All patients were administrated a basic antiplatelet and lipid-lowering therapy. Among the patients, 86 received additional NBP administration for 30 days, whereas 84 received only basic therapy (the control). The level of circulating EPCs (marked with CD34(+)/CD133(+)/KDR(+)) was determined by flow cytometry at baseline and days 7, 14, and 30 after therapy. Impairment of neurological function was evaluated by the National Institutes of Health Stroke Scale (NIHSS) on days 7, 14, 30, and 90 after therapy. The association between the increased level of circulating EPCs and improvement of NIHSS score was evaluated by Pearson analysis. The clinical outcome was evaluated by modified Rankin Scale (mRS) on day 90. During the observation period, any adverse events related to drugs were reported.. The levels of circulating EPCs on days 14 and 30 were significantly higher in the NBP group than in the control group. In contrast, NIHSS score was notably lower in NBP group on day 14, 30 and day 90. Pearson correlation analysis revealed a significant association between the increased level of EPCs and improvement of NIHSS score. Also, the mRS score in the NBP group was lower on day 90. Importantly, the reported adverse events in the 2 groups were comparable.. NBP significantly increases the circulating level and improves clinical outcome in patients with AIS.

Topics: Aged; Antigens, CD; Benzofurans; Brain Ischemia; Cell Movement; Diffusion Magnetic Resonance Imaging; Endothelial Progenitor Cells; Female; Follow-Up Studies; Humans; Male; Middle Aged; Platelet Aggregation Inhibitors; Retrospective Studies; Severity of Illness Index; Statistics as Topic; Stroke; Time Factors; Vascular Endothelial Growth Factor Receptor-2

Synapse impairment in the Alzheimer's disease (AD) brain is an early event leading to cognitive dysfunction. Most oxidative stress localizes to the synapse, and synapse loss is the basis of cognitive decline in AD. Dl-3-n-butylphthalide (Dl-NBP), a small molecule compound has been shown to ameliorate oxidative stress. We evaluated the effects of a 5-month oral delivery with Dl-NBP on oxidative stress and cognitive function in APP/PS1 transgenic mice. Dl-NBP treatment reduced oxidative stress in the APP/PS1 mouse brain and alleviated learning and memory deficits. Dl-NBP supplementation meliorated synaptic plasticity, diminished soluble amyloid beta and amyloid beta oligomer in the APP/PS1 mouse brain. Dl-NBP administration caused an increase of cyclic adenosine monophosphate-response element binding protein (CREB)-binding protein (CBP)-associated Ser133-phosphorylated CREB (p-CREB) protein. Chromatin immunoprecipitation analysis revealed that Dl-NBP increased the recruitment of CBP to the promoters of best-characterized genes downstream of nuclear factor erythroid 2-related factor 2, nicotinamide adenine dinucleotide phosphate (NADPH) quinone oxidoreductase 1, and γ-glutamylcysteine synthetase modifier subunit. We demonstrate that the Dl-NBP-triggered upregulation of antioxidant defenses is involved in the enhancement of cross talk between CREB and nuclear factor erythroid 2-related factor 2 via CBP. Our results suggest that Dl-NBP may be a useful agent for the treatment of AD.

Topics: Administration, Oral; Alzheimer Disease; Amyloid beta-Peptides; Animals; Antioxidants; Benzofurans; Brain; Cognition; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Male; Mice, Inbred C57BL; Mice, Transgenic; Neuronal Plasticity; NF-E2-Related Factor 2; Oxidative Stress; Up-Regulation

Vascular smooth muscle cells (VSMCs) played an important role in vascular remodeling. dl-3-n-butylphthalide (NBP) was extracted as a pure component from seeds of Apium graveolens Linn (Chinese celery) for protecting neurons activity, but the role of NBP on VSMCs was not clearly clarified.. Cell proliferation was measured by MTS and flow cytometry. Western blot analysis and transmission electron microscopy were performed to analyze the relative protein expression and autophagosome. Moreover, the autophagic inhibitor and β-catenin inhibitor were used to evaluate the effects of NBP on autophagy and the function of β-catenin on cell proliferation respectively.. NBP significantly suppressed platelet derived growth factor-BB (PDGF-BB)-stimulated VSMC proliferation, and the inhibitory effects of NBP on proliferation were caused by inducing autophagy. In addition, the inhibitory effects of NBP on proliferation were associated with the β-catenin signaling pathway. Moreover, β-catenin overexpression reversed the induction effect of NBP on autophagy and the β-catenin inhibitor JW74 enhanced these effects.. Our findings demonstrated that NBP protected VSMC from PDGF-BB-stimulated proliferation by inducing autophagy through suppression of the β-catenin signaling pathway, confirming the induction of autophagy might be a therapeutic strategy for use in the proliferative cardiovascular diseases.

Topics: Animals; Autophagosomes; Autophagy; Becaplermin; Benzofurans; beta Catenin; Cell Cycle; Cell Proliferation; Cells, Cultured; Male; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Oxadiazoles; Proto-Oncogene Proteins c-sis; Rats; Triazoles

Although Butylphthalide (BP) has protective effects that reduce ischemia-induced brain damage and neuronal cell death, little is known about the precise mechanisms occurring during cerebral ischemia/reperfusion (I/R). Therefore, the aim of this study was to investigate the neuroprotective mechanisms of BP against ischemic brain injury induced by cerebral I/R through inhibition of the c-Jun N-terminal kinase (JNK)-Caspase3 signaling pathway. BP in distilled non-genetically modified Soybean oil was administered intragastrically three times a day at a dosage of 15 mg/(kg day) beginning at 20 min after I/R in Sprague-Dawley rats. Immunohistochemical staining and Western blotting were performed to examine the expression of related proteins, and TUNEL-staining was used to detect the percentage of neuronal apoptosis in the hippocampal CA1 region. The results showed that BP could significantly protect neurons against cerebral I/R-induced damage. Furthermore, the expression of p-JNK, p-Bcl2, p-c-Jun, FasL, and cleaved-caspase3 was also decreased in the rats treated with BP. In summary, our results imply that BP could remarkably improve the survival of CA1 pyramidal neurons in I/R-induced brain injury and inhibit the JNK-Caspase3 signaling pathway.

Topics: Animals; Apoptosis; Benzofurans; Brain Ischemia; Caspase 3; JNK Mitogen-Activated Protein Kinases; Male; MAP Kinase Signaling System; Neurons; Rats, Sprague-Dawley; Reperfusion Injury; Signal Transduction

To search for novel anti-ischemic stroke agents with higher potency than a known drug 3-n-butylphthalide (NBP), a series of ring-opened derivatives of NBP bearing both nitric oxide (NO) and hydrogen sulfide (H2S)-donating moieties (NO/H2S-NBP) (8a-8o) were designed, synthesized, and biologically evaluated. The most active compound 8d was more potent than NBP and the corresponding H2S-NBP 10 or NO-NBP 13 in inhibition of the ADP-induced platelet aggregation in vitro. In addition, 8d produced moderate levels of NO and H2S, which could be beneficial for improving cardiovascular and cerebral circulation. More importantly, in a rat model of transient focal cerebral ischemia, oral treatment with 8d improved neurobehavioral function, reduced the infarct brain size and brain-water content, and enhanced the levels of brain antioxidant SOD, GSH and GSH-Px but diminished the level of oxidant MDA. These protective effects of 8d against the ischemia/reperfusion (I/R)-related brain damage were greater than that of NBP, suggesting that 8d may be a promising agent for further investigation.

Topics: Animals; Antioxidants; Benzofurans; Drug Discovery; Male; Rats; Stroke

5d, a novel analogue of the racemic 3-n-butylphthalide (NBP), has been reported for its free radical scavenging activity in vitro and preventive neuroprotection in vivo. Nevertheless, the mechanism by which 5d attenuated ischemia/reperfusion (I/R) injury is still unknown. Our results showed that 5d significantly increased CK2 activity as well as CK2α and 2α' protein levels after I/R injury. Besides, 5d suppressed the translocation of cytosolic p47phox and Rac1 to the membrane, decreased NOX4 expression and ROS generation. Furthermore, 5d blocked the dissociation between CK2α and Rac1 so as to decrease NADPH oxidase activity. Based on these findings, we propose that the neuroprotective effect of 5d is due to an increase of CK2 activity, which blocks I/R-induced dissociation between CK2α and Rac1, decreases NADPH oxidase activity, inhibits ROS production and finally realizes the neuroprotection of I/R. These findings point to that 5d might be considered an attractive candidate for further studies in ischemic stroke.

Topics: Animals; Benzofurans; Brain; Casein Kinase II; Cell Membrane; Cell Survival; Cytosol; Gene Expression Regulation, Enzymologic; Infarction, Middle Cerebral Artery; Male; NADPH Oxidases; Neurons; Neuroprotective Agents; Oxidative Stress; rac1 GTP-Binding Protein; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion Injury; RNA Interference

Angelica sinensis (Oliv.) hexane extract (AHE) has been reported as a proven and impressive repellent against laboratory-reared female Aedes aegypti mosquitoes. With the aim of promoting products of plant origin as a viable alternative to conventional synthetic substances, this study was designed to transform AHE-based repellents for exploitable commercial production by enhancing their efficacy and assessing their physical and biological stability as well as repellency against mosquitoes under laboratory and field conditions.. The chemical profile of AHE was analyzed by qualitative gas chromatography-mass spectrometry (GC-MS) technique. AHE was supplemented with vanillin, as a fixative, and then investigated for repellency and comparison to the standard synthetic repellent, DEET, under both laboratory and field conditions. Determination of physical and biological stability as a repellent was carried out after keeping AHE samples under varying temperatures and for different storage times.. GC-MS analysis revealed that AHE contained at least 21 phytochemical compounds, constituting 95.74 % of the total content, with the major constituent of 3-N-butylphthalide (66.67 %). Ethanolic formulations of AHE and DEET showed improvement of repellency in a dose-dependent manner when vanillin was added in laboratory assessment. While 5-25 % AHE alone provided median complete-protection times of 2.0-6.5 h against Ae. aegypti, these times were increased to 4.0-8.5 h with a combination of AHE and 5 % vanillin (AHEv). Protection times against Ae. aegypti were extended from 2.25 to 7.25 h to 4.25-8.25 h when 5-25 % DEET was combined with 5 % vanillin (DEETv). In determining stability, all stored AHE samples exhibited similar characteristics such as liquid phases with aromatic odor comparable to those of fresh preparations. Furthermore, repellent activity of stored AHE samples lasted for at least six months, with varied efficacy (4.5-10.0 h) against Ae. aegypti. Field trials revealed strong repellency from both 25 % AHEv and 25 % DEETv, with complete protection (100 %) against a wide range of local mosquito populations. A total of 5,718 adult female mosquitoes, with the most predominant being Culex quinquefasciatus (41.47 %), Armigeres subalbatus (41.13 %), and Culex vishnui (10.53 %), was collected during field applications. No local skin reaction or other allergic responses was observed during both laboratory and field study periods.. Angelica sinensis proved to have not only impressive repellency against both laboratory Ae. aegypti and a wide range of natural mosquito populations, but also relative stability in physical and biological performance.

Topics: Angelica sinensis; Animals; Benzofurans; Culicidae; Female; Humans; Insect Repellents; Linoleic Acid; Male; Molecular Structure; Phthalic Anhydrides; Plant Extracts; Thailand

Recent studies have shown that the two-pore-domain potassium channel TREK-1 is involved in the proliferation of neural stem cells, astrocytes and human osteoblasts. In this study, we investigated how TREK-1 affected the proliferation of Chinese hamster ovary (CHO) cells in vitro.. A CHO cell line stably expressing hTREK-1 (CHO/hTREK-1 cells) was generated. TREK-1 channel currents in the cells were recorded using whole-cell voltage-clamp recording. The cell cycle distribution was assessed using flow cytometry analysis. The expression of major signaling proteins involved was detected with Western blotting.. CHO/hTREK-1 cells had a high level of TREK-1 expression, reached up to 320%±16% compared to the control cells. Application of arachidonic acid (10 μmol/L), chloroform (1 mmol/L) or etomidate (10 μmol/L) substantially increased TREK-1 channel currents in CHO/hTREK-1 cells. Overexpression of TREK-1 caused CHO cells arresting at the G1 phase, and significantly decreased the expression of cyclin D1. The TREK-1 inhibitor l-butylphthalide (1-100 μmol/L) dose-dependently attenuated TREK-1-induced G1 phase cell arrest. Moreover, overexpression of TREK-1 significantly decreased the phosphorylation of Akt (S473), glycogen synthase kinase-3β (S9) and cAMP response element-binding protein (CREB, S133), enhanced the phosphorylation of p38 (T180/Y182), but did not alter the phosphorylation and expression of signal transducer and activator of transcription 3 (STAT3).. TREK-1 overexpression suppresses CHO cell proliferation by inhibiting the activity of PKA and p38/MAPK signaling pathways and subsequently inducing G1 phase cell arrest.

Topics: Animals; Arachidonic Acid; Benzofurans; Blotting, Western; Cell Proliferation; Chloroform; CHO Cells; Cricetinae; Cricetulus; Cyclic AMP-Dependent Protein Kinases; Etomidate; Flow Cytometry; G1 Phase Cell Cycle Checkpoints; Humans; Membrane Potentials; p38 Mitogen-Activated Protein Kinases; Patch-Clamp Techniques; Potassium Channels, Tandem Pore Domain; Signal Transduction; Transfection

Our previous studies showed that L-3-n-butylphthalide (L-NBP), an extract from seeds of Apium graveolens Linn (Chinese celery), improved cognitive ability in animal models of cerebral ischemia, vascular dementia, and Alzheimer's disease (AD). It is well known that cognitive deficit of AD is caused by synaptic dysfunction. In this study, we investigated the effect of L-NBP on hippocampal synaptic function in APP/PS1 AD transgenic mice and related mechanisms.. Eighteen-month-old APP/PS1 transgenic (Tg) mice were administrated 15 mg/kg L-NBP by oral gavage for 3 months. Synaptic morphology and the thickness of postsynaptic density (PSD) in hippocampal neurons were investigated by electron microscope. The dendritic spines, Aβ plaques, and glial activation were detected by staining. The expressions of synapse-related proteins were observed by Western blotting.. L-NBP treatment significantly increased the number of synapses and apical dendritic thorns and the thickness of PSD, increased the expression levels of synapse-associated proteins including PSD95, synaptophysin (SYN), β-catenin, and GSK-3β, and attenuated Aβ plaques and neuroinflammatory responses in aged APP/PS1 Tg mice.. L-NBP may restore synaptic and spine function in aged APP Tg mice through inhibiting Aβ plaques deposition and neuroinflammatory response. Wnt/β-catenin signaling pathway may be involved in L-NBP-related restoration of synaptic function.

Topics: Alzheimer Disease; Amyloid beta-Protein Precursor; Animals; Benzofurans; beta Catenin; Dendritic Spines; Disease Models, Animal; Disks Large Homolog 4 Protein; Gene Expression Regulation; Glycogen Synthase Kinase 3 beta; Hippocampus; Humans; Intracellular Signaling Peptides and Proteins; Male; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Neuroprotective Agents; Presenilin-1; Synapses

N-Butylphthalide (NBP) has been known to have potential neuroprotective effects in Alzheimer's disease and stroke animal models. Hepatocyte-growth factor (HGF), with strong angiogenic properties, exerted protective role in brain injury. The present study was aimed to investigate the possible anti-inflammatory effects of NBP on the brain injury of rats with cerebral ischemia reperfusion (IR) and astrocytes activation induced by lipopolysaccharide (LPS) treatment. Our results showed that cerebral IR induced brain damage with down-regulation of HGF and astrocytes activation. NBP treatment significantly increased HGF expression and activated cMet/PI3K/AKT signaling pathway, stimulating mTOR activity and suppressing apoptosis in brain tissues. Also NBP inhibited pro-inflammatory cytokines expression, including IL-6, IL-1β, and TNFα, via TLR4/NF-κB suppression. Anti-HGF treatment enhanced TLR4 expression while HGF could suppress TLR4 activation and its down-streaming signals, attenuating inflammation finally. Notably, NBP up-regulated HGF and down-regulated TLR4 expression significantly in the astrocytes combined with the treatment of TLR4 inhibitor than the cells only treated with TLR4 inhibitor, suggesting that NBP could further suppress TLR4 activation, suggesting that NBP might impede TLR4 through up-regulating HGF expression. These results suggested that NBP treatment significantly ameliorated cerebral IR-induced brain injury by inhibiting TLR4/NF-κB-associated inflammation regulated by HGF.

Topics: Animals; Astrocytes; Benzofurans; Brain Ischemia; Cell Survival; Hepatocyte Growth Factor; Humans; Inflammation; Lipopolysaccharides; Male; NF-kappa B; Rats, Sprague-Dawley; Reperfusion Injury; Signal Transduction; Toll-Like Receptor 4; Up-Regulation

In this study we investigated the effect of Dl-3-n-butylphthalide (NBP), a clinically used drug for stroke patients in China, on the recovery following cardiac arrest and resuscitation in rats. Male Wistar rats (3-month old) underwent cardiac arrest (12 min) and resuscitation. Rats were randomly assigned to the following groups: sham non-arrested group, vehicle group (vehicle-treated, 7 days before cardiac arrest and 4 days post-resuscitation), NBP pre-treated group (NBP-treated, 7 days before cardiac arrest), and NBP post-treated group (NBP-treated, 4 days post-resuscitation). Overall survival rates and hippocampal neuronal counts were determined in each group at 4 days post-resuscitation. Results showed that NBP pre-treated group (80 %) and NBP post-treated group (86 %) had significantly higher survival rates compared to that of the vehicle group (50 %). At 4 days of recovery, only about 20 % of hippocampal neurons were preserved in the vehicle group compared to the sham non-arrested group. The hippocampal CA1 cell counts in the NBP pre-treated group and NBP post-treated group were significantly higher than the counts in the vehicle group, about 50-60 % of the counts of non-arrested rats. The data suggest that NBP has both preventive and therapeutic effect on improving outcome following cardiac arrest and resuscitation, and NBP might be a potential early phase treatment for patients recovered from cardiac arrest and resuscitation.

Topics: Animals; Benzofurans; CA1 Region, Hippocampal; Disease Models, Animal; Heart Arrest; Male; Neurons; Neuroprotective Agents; Rats, Wistar; Recovery of Function; Resuscitation; Time Factors

Methamphetamine (Meth) abuse causes neural injury in the brain. There are no efficacious therapies available to treat Meth-induce neural injury. The present study intended to test the therapeutic potential of dl-3-n-butylphthalide (NBP), a chemical compound extracted originally from the seeds of Chinese Celery, in the amelioration and prevention of Meth-induced neural injury.. Experiments were carried out on SH-SY5Y cells. Neuronal injury and apoptotic cell death were detected by MTT assay and analysis of nuclear morphology. Intracellular reactive oxygen species (ROS) was evaluated by dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay and protein expression levels of the apoptosis-related cleaved caspase-3, bcl2 and Bax were determined by western blotting.. Treatment of SH-SY5Y cells with Meth induced cell injury and apoptosis. NBP attenuated Meth-associated cell injury and apoptosis via blockage of Meth-mediated upregulation of intracellular ROS production and inhibition of Meth-induced decrease of cleaved caspase-3/caspase-3 and Bcl-2/Bax ratios.. The results presented in this study indicate that NBP may have therapeutic benefits in the treatment of Meth-induced neuronal injury in the central nervous system.

Topics: Benzofurans; Cell Line, Tumor; Humans; Methamphetamine; Neuroblastoma; Neurotoxicity Syndromes; Reactive Oxygen Species; Signal Transduction

In the present study, a series of novel nitric oxide-hydrogen sulfide releasing derivatives of (S)-3-n-butylphthalide ((S)-NBP) were designed, synthesized, and evaluated as potential antiplatelet agents. Compound NOSH-NBP-5 displayed the strongest activity in inhibiting the arachidonic acid (AA)- and adenosine diphosphate (ADP)-induced platelet aggregation in vitro, with 3.8- and 7.0-fold more effectiveness than (S)-NBP, respectively. Furthermore, NOSH-NBP-5 could release moderate levels of NO and H

Topics: Animals; Benzofurans; Humans; Hydrogen Sulfide; Male; Molecular Structure; Nitric Oxide; Platelet Aggregation; Platelet Aggregation Inhibitors; Rabbits; Rats; Rats, Sprague-Dawley; Thrombosis

Diabetes mellitus is associated with rapid cognitive decline. Currently, there is no effective treatment for cognitive dysfunction induced by diabetes. L-3-n-Butylphthalide (L-NBP) is a nerve protective drug extracted from seeds of celery, which has been proved to improve learning and memory in vascular dementia animal models by improving microcirculation, protecting mitochondria and increasing long-term potentiation (LTP). NR2B, one of the subunits of N-methyl-D-aspartate receptor, has been proved to be an important factor for the formation of LTP. The study aimed to investigate the role of NR2B in cognitive dysfunction in the rats with type 1 diabetes and define the protective effects of L-NBP on cognition. A rat model of type 1 diabetes was established by a single intraperitoneal injection of streptozotocin at 60 mg/kg. Animals were randomly allocated to four groups: normal control (NC); diabetic control (DC); diabetic + low L-NBP (DL, administered L-NBP 60 mg/kg per day for 12 weeks); and diabetic + high L-NBP (DH, administered L-NBP 120 mg/kg per day, for 12 weeks). After 12 weeks, cognitive and memory changes were investigated in the Morris water maze. The expression of NR2B was assessed by real-time polymerase chain reaction, Western blotting, and immunohistochemistry. Our results indicated that the escape latency was significantly increased and the number of crossing platform was significantly decreased in DC group compared to NC group. Also, the expression of NR2B was significantly declined in DC group. However, compared to DC group, the expression of NR2B of the L-NBP-treated groups was significantly increased and the escape latency was shortened with the DH group being the most obvious. Therefore, L-NBP can improve the cognitive function by up-regulating the expression of NR2B in STZ-diabetic rats, which may provide the direction for future diabetic encephalopathy therapy.

Topics: Animals; Benzofurans; Blood Glucose; Body Weight; Cognition; Diabetes Mellitus, Experimental; Gene Expression Regulation; Hippocampus; Male; Maze Learning; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; RNA, Messenger

Previous studies have demonstrated that a natural product of celery seeds, 3‑n‑butylphthalide (NBP), has significant antihypertensive effects that are widely utilized in Chinese traditional medicine. The present study aimed to investigate the effects of NBP on hypertensive nephropathy, as well as the mechanisms underlying this disease in spontaneously hypertensive rats (SHRs). SHRs were treated orally with saline, NBP (15 or 30 mg/kg) or losartan (10 mg/kg) daily for 20 weeks, during which time blood pressure was measured every four weeks. At the end of the 20‑week treatment, blood and urine samples were collected for biochemical analysis, and kidney tissues were obtained for histopathological analysis and immunohistochemistry. Enzyme‑linked immunosorbent assays and western blotting were used to analyze the expression of transforming growth factor (TGF)‑β1 in blood and kidney tissues, respectively. The results showed that NBP effectively attenuated progression of hypertensive nephropathy by decreasing urinary albumin excretion and blood urea nitrogen levels. It significantly decreased blood pressure (although less markedly than losartan) and the incidence of glomerulosclerosis. In addition, it alleviated tubular impairment and significantly decreased oxidative stress, as well as the expression of pro‑inflammatory cytokines and TGF-‑β1 in kidney tissues. In conclusion, the results suggested that NBP may slow the progression of hypertensive nephropathy by a variety of mechanisms.

Topics: Albumins; Animals; Antihypertensive Agents; Benzofurans; Blood Pressure; Blood Urea Nitrogen; Drugs, Chinese Herbal; Enzyme-Linked Immunosorbent Assay; Hypertension, Renal; Immunohistochemistry; Interleukin-6; Kidney Glomerulus; Losartan; Male; NADPH Oxidases; Nephritis; Neuroprotective Agents; NF-kappa B; Rats; Rats, Inbred SHR; Transforming Growth Factor beta1

A high-throughput, rapid, sensitive, environmentally friendly, and economical supercritical fluid chromatography with triple quadruple mass spectrometry method was established and validated for the first time to determine a cerebral stroke treatment drug named 3-n-butylphthalide in dog plasma. Plasma samples were prepared by protein precipitation with methanol and the analytes were eluted on an ACQUITY UPC(2TM) HSS-C(18) SB column (3 × 100 mm, 1.8 μm) maintained at 50°C. The mobile phase comprised supercritical carbon dioxide/methanol (90:10, v/v) at a flow rate of 1.5 mL/min, the compensation solvent was methanol at a flow rate of 0.2 mL/min and the total run time was 1.5 min per sample. The detection was carried out on a tandem mass spectrometer with an electrospray ionization source. Calibration curves were linear over the concentration range of 1.02-1021.00 ng/mL (r(2) ≥ 0.993) with the lower limit of quantification of 1.02 ng/mL. The intra- and inter-day precision values were below 15% and the accuracy was from 97.90 to 103.70% at all quality control levels. The method was suitable for a pharmacokinetic study of 3-n-butylphthalide in beagle dogs.

Topics: Animals; Benzofurans; Biological Availability; Chromatography, Supercritical Fluid; Dogs; Drugs, Chinese Herbal; Male; Mass Spectrometry

Aberrant platelet activation plays a critical role in the pathogenesis of heart attack and stroke. DL-3-n-butylphthalide (NBP) has been approved in China to treat stroke with multiple mechanisms. The anti-stroke effects of NBP may be related to its antiplatelet effects reported in rats in addition to its antioxidative, antiapoptotic, and angiogenic effects. However, the effects and the underlying mechanisms of NBP on human platelets are not yet clear. In this study, we found that NBP concentration-dependently inhibited human platelet aggregation and ATP release induced by ADP, thrombin, U46619, arachidonic acid, or collagen. NBP also inhibited PAC-1 binding induced by ADP or thrombin and platelet spreading on immobilized fibrinogen. NBP reduced TXA2 synthesis induced by thrombin or collagen via inhibiting cPLA2 phosphorylation, concomitantly with a marked decrease in intracellular calcium mobilization. Moreover, NBP also inhibited human platelet phosphodiesterase (PDE) and elevated 3,5-cyclic adenosine monophosphate level in platelets. In conclusion, NBP significantly inhibits human platelet activation via inhibition of cPLA2-mediated TXA2 synthesis and PDE, and may be effective as an antiplatelet drug to treat other arterial thrombotic diseases.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Adenosine Diphosphate; Benzofurans; Blood Platelets; Calcium; Collagen; Cyclic AMP; Humans; Phospholipases A2, Cytosolic; Phosphoric Diester Hydrolases; Phosphorylation; Platelet Activation; Platelet Aggregation Inhibitors; Thrombin; Thromboxane A2

Carbon monoxide (CO) intoxication is one of the most common types of poisoning worldwide, and may result in neuropathologic sequelae, yet its pathogenesis is not clear and there is no optimal management strategy for patients with CO poisoning. In this study, the rat model of CO poisoning was established in a hyperbaric chamber by CO exposure. Rats were administered orally N-Butylphthalide (NBP) at a dose of 1 ml/100g. Neuronal apoptosis was assessed by TUNEL stain and flow cytometry. The expressions of neurite outgrowth inhibitor (Nogo), myelin-associated glycoprotein (MAG) and Nogo receptor-1 (NgR1) were observed in rat brain tissue by immunohistochemistry and double immunofluorescence staining. As we expected, CO poisoning could start the mechanism of apoptosis. The number of apoptotic cells and the early neuronal apoptosis percentage (EAR) were significantly increased at 1 day, 3 day after CO exposure. NBP treatment obviously reduce neuronal apoptosis and the EAR (P<0.05). CO poisoning could induce Nogo, MAG and NgR1 expressions. The increased Nogo, MAG and NgR1 proteins were still observed at 4 week after CO poisoning. NBP could significantly reduce the levels of Nogo and NgR1 proteins. Then we suspected that the expressions of Nogo, MAG and NGR1 proteins might be associated with brain injury and demyelination induced by CO poisoning. NBP might inhibit neuronal apoptosis and the EAR, down-regulate the expressions of Nogo and NgR1 proteins (but not MAG), and play a neuro-protective role in brain damage after acute CO poisoning.

Topics: Animals; Apoptosis; Benzofurans; Brain; Carbon Monoxide Poisoning; GPI-Linked Proteins; Male; Myelin Proteins; Myelin-Associated Glycoprotein; Neuroprotective Agents; Nogo Proteins; Nogo Receptor 1; Rats, Sprague-Dawley; Receptors, Cell Surface

L-3-n-butylphthalide molecularly imprinted polymers (MIPs) were synthesized using l-3-n-butylphthalide as template molecule, acrylamide as functional monomer, ethylene glycol dimethacrylate as cross-linking agent, and acetone as the porogenic solvent through precipitation polymerization. The non-imprinted polymers (NIPs) were prepared with the same procedure, but with the absence of template molecule. The optimum preparation conditions of the MIPs such as the functional monomer, the porogenic solvent, the molar ratio of the template to the functional monomer and the molar ratio of the template to the cross-linker were investigated in detail. Prior to the polymerization, the molecular simulation with the computer-aided design was used to help choose a suitable polymerization porogen for the molecularly imprinted pre-assembled system and study the interactions between l-NBP and the functional monomers. The synthesized polymers were characterized with FTIR and SEM to observe their structures as well as the morphologies, and their adsorption properties were respectively evaluated by static and dynamic adsorption as well as selectivity experiments. Scatchard analyses revealed that there were high and low affinity sites formed in the MIPs, which elucidated good affinity to l-NBP in the ethanol system. The adsorption capacity of the MIPs for l-NBP was 3.561 mg g(-1), with an imprinting factor (α) of 2.321 when compared with that of the NIPs. Scatchard analysis illustrated that the binding sites with affinity for l-3-n-butylphthalide molecules were formed in the prepared MIPs.

Topics: Adsorption; Benzofurans; Chemical Precipitation; Molecular Imprinting; Polymerization; Solid Phase Extraction

Carbon monoxide (CO) is the leading cause of death by poisoning all over the world and may result in neuropathologic changes and cognitive and neurologic sequelae, yet little is known regarding its outcomes. The present study aimed to evaluate the neuroprotective effects of N-butylphthalide (NBP) against brain damage after acute CO poisoning. The animal model of CO poisoning was established by exposed to 1000 ppm CO in air for 40 min and then to 3000 ppm for another 20 min. RT-PCR was used to assess the expressions of apoptosis-associated genes Bcl-2 mRNA and Bax mRNA. Mitochondrial membrane potential (MMP) was detected by fluorescent probe JC-1. Immunohistochemistry stain and Western blot assay were used to evaluate the expression levels of Kelch-like ECH-associated protein 1 (Keapl), nuclear factor erythroid 2-related factor 2 (Nrf-2) and. quinone oxidoreductase 1(NQO-1). CO poisoning could increase the levels of Bcl-2 mRNA and Bax mRNA expressions, and obviously decrease the MMP of cells. NBP treatment could maintain the high MMP, significantly up-regulate Bcl-2 mRNA and down-regulate Bax mRNA expression, and the ratio of Bcl-2 mRNA/Bax mRNA expressions was higher than that in the CO poisoning group (P<0.05). CO poisoning could start oxidative stress response. The expressions of Keap1, Nrf-2 and NQO-1 proteins significantly increased at 1, 3 and 7 day after NBP administration as compared with the CO poisoning group (P<0.01). These findings suggest that N-butylphthalide may protect mitochondrial function, balance the expressions of anti-apoptosis genes and pro-apoptosis genes, be in part associated with activation of Keap1-Nrf-2/antioxidant response element (ARE) signaling pathway, and play a neuroprotective role in brain damage after acute CO poisoning.

Topics: Animals; Apoptosis; Benzofurans; Brain; Carbon Monoxide Poisoning; Intracellular Signaling Peptides and Proteins; Kelch-Like ECH-Associated Protein 1; Membrane Potential, Mitochondrial; NF-E2-Related Factor 2; Rats; Signal Transduction

Compound 10b is a hybrid molecule of edaravone and a ring-opening derivative of 3-n-butylphthalide (NBP). The aim of this study was to examine the effects of compound 10b on brain damage in rats after focal cerebral ischemia.. SD rats were subjected to 2-h-middle cerebral artery occlusion (MCAO). At the onset of reperfusion, the rats were orally treated with NBP (60 mg/kg), edaravone (3 mg/kg), NBP (60 mg/kg)+edaravone (3 mg/kg), or compound 10b (70, 140 mg/kg). The infarct volume, motor behavior deficits, brain water content, histopathological alterations, and activity of GSH, SOD, and MDA were analyzed 24 h after reperfusion. The levels of relevant proteins in the ipsilateral striatum were examined using immunoblotting.. Administration of compound 10b (70 or 140 mg/kg) significantly reduced the infarct volume and neurological deficits in MCAO rats. The neuroprotective effects of compound 10b were more pronounced compared to NBP, edaravone or NBP+edaravone. Furthermore, compound 10b significantly upregulated the protein levels of the cytoprotective molecules Bcl-2, HO-1, Nrf2, Trx, P-NF-κB p65, and IκB-α, while decreasing the expression of Bax, caspase 3, caspase 9, Txnip, NF-κB p65, and P-IκB-α.. Oral administration of compound 10b effectively attenuates rat cerebral ischemia injury.

Topics: Animals; Antioxidants; Antipyrine; Apoptosis; Benzofurans; Brain; Edaravone; Infarction, Middle Cerebral Artery; Male; Neuroprotective Agents; NF-kappa B; Rats, Sprague-Dawley; Signal Transduction; Water

Demyelinating diseases is common in neurology department, but its treatment is still not clear. A 40-year-old male who was addicted in heroin was hospitalized and presented worsening altered mental status in hospital. Brain magnetic resonance imaging (MRI) revealed a symmetrical diffuse long T1 and long T2 time abnormalities in the white matter of cerebral hemispheres which indicated demyelination. High-dose intravenous methylprednisolone was not effective as expected. However, after treatment with intravenous Dl-3-n-butylphthalide for 7 days, the patient's symptoms and features of electroencephalogram (EEG) and MRI improved. Hence, ethidium bromide was used in a rat model to evaluate the positive effects of Dl-3-n-butylphthalide in demyelinating diseases. The results indicated that Dl-3-n-butylphthalide prevented white matter from demyelinating by regulating apoptosis through multiple approaches.

Topics: Adult; Animals; Benzofurans; Brain; Demyelinating Diseases; Disease Models, Animal; Electroencephalography; Humans; Magnetic Resonance Imaging; Male; Methylprednisolone; Radiography; Rats, Sprague-Dawley; Treatment Outcome

To develop novel anti-ischemic stroke agents with better therapeutic efficacy and bioavailability, we designed and synthesized a series of 3-alkyl-2,3-dihydro-1H-isoindol-1-ones compounds (3a-i) derivatives, one of which (3d) exhibited the strongest inhibitory activity for the adenosine diphosphate-induced and arachidonic acid-induced platelet aggregation. This activity is superior to that of 3-n-butylphthalide and comparable with aspirin and edaravone. Meanwhile, 3d not only exhibited a potent activity in scavenging free radicals and improving the survival of HT22 cells against the reactive oxygen species-mediated cytotoxicity in vitro but also significantly attenuated the ischemia/reperfusion-induced oxidative stress in ischemic rat brains. Results from transient middle cerebral artery occlusion and permanent middle cerebral artery occlusion model, indicated that 3d could significantly reduce infarct size, improve neurobehavioral deficits, and prominently decrease attenuation of cerebral damage. Most importantly, 3d possessed a very high absolute bioavailability and was rapidly distributed in brain tissue to keep high plasma drug concentration for the treatment of ischemic strokes. In conclusion, our findings suggest that 3-alkyl-2,3-dihydro-1H-isoindol-1-ones, a novel series of compounds, might be candidate drugs for the treatment of acute ischemic strokes, and 3d may be a promising therapeutic agent for the primary and secondary prevention of ischemic stroke.

Topics: Animals; Antioxidants; Antipyrine; Behavior, Animal; Benzofurans; Biological Availability; Brain; Carrageenan; Cell Line; Disease Models, Animal; Edaravone; Fibrinolytic Agents; Infarction, Middle Cerebral Artery; Isoindoles; Male; Mice; Molecular Structure; Neuroprotective Agents; Oxidative Stress; Platelet Aggregation; Platelet Aggregation Inhibitors; Rats, Sprague-Dawley; Reperfusion Injury; Structure-Activity Relationship; Thrombosis; Tissue Distribution

Fourteen hybrids (10a-g, 11a-g) of 3-n-butylphthalide (NBP) and edaravone (Eda) analogues have been designed and synthesized as potential anti-ischemic stroke agents. In vitro biological studies showed that compounds 10d and 10g exhibited more potent anti-platelet aggregation than ticlopidine (Ticlid), aspirin (ASP) and NBP. Compound 10g more significantly prevented H2O2-mediated neuronal cell (PC12) death than NBP, Eda or NBP together with Eda. Meanwhile, 10g also possessed potent radical scavenging effects on hydroxyl radical (˙OH) and superoxide anion radical (˙O2(-)). Our findings may provide new insights into the development of these hybrids, like 10g, for the intervention of ischemic stroke.

Topics: Animals; Antipyrine; Benzofurans; Cell Line; Drug Design; Edaravone; Free Radical Scavengers; Humans; Hydrogen Peroxide; Neurons; Neuroprotective Agents; Platelet Aggregation; Platelet Aggregation Inhibitors; Rats; Stroke

This study investigated whether anticerebral ischemia new drug, l-3-n-butylphthalide (l-NBP), improved behavioral recovery and enhanced hippocampal neurogenesis after cerebral ischemia in rats.. The middle cerebral artery of rats was blocked for 2 h. The daily oral administrations of 30 mg/kg l-NBP or vehicle were begun from the second day until the rats were sacrificed. L-NBP treatment markedly increased 5-bromo-2'-deoxyuridine (BrdU)-positive cells in the hippocampal dentate gyrus (DG) of injured hemisphere on day 28 after ischemia. The amount of newborn cells and newly mature neurons was also increased. The expressions of growth-associated protein-43 and synaptophysin were significantly elevated in l-NBP-treated rats. However, l-NBP markedly reduced the percentage of BrdU(+) /GFAP(+) cells. Additionally, the levels of catalytical subunit of protein kinase A (PKA), protein kinase B (Akt), and cAMP response element-binding protein (CREB) were significantly increased, and the activation of the signal transducer and activation of transcription 3 (STAT3) and the expressions of cleaved caspase-3 and Bax were obviously inhibited by l-NBP. Consequently, l-NBP attenuated the behavioral dysfunction.. It first demonstrates that l-NBP may improve the behavioral outcome of cerebral ischemia by promoting neurogenesis and neuroplasticity. Activation of CREB and Akt and inhibition of STAT3 signaling might be involved in.

Topics: Administration, Oral; Animals; Apoptosis; Benzofurans; Brain Ischemia; Cell Survival; Cognition; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Hippocampus; Infarction, Middle Cerebral Artery; Male; Neural Stem Cells; Neurogenesis; Neuronal Plasticity; Neuroprotective Agents; Proto-Oncogene Proteins c-akt; Rats, Sprague-Dawley; Recovery of Function; STAT3 Transcription Factor; Synapses

The work aims to study the drug metabolizing enzymes involved in the metabolism of butylphthalide and evaluate the induction and inhibition activities of butylphthalide on CYP450 isoenzymes by using in vitro (liver microsome incubation system of rats and human) and in vivo (CYP induced model of rats) method. Butylphthalide was incubated with selective inhibitors of CYP450, and its metabolic rate was determined to identify the metabolizing isoenzymes of NBP in rat (normal and induced rats) and human liver microsomes. The in vitro inhibition effect of butylphthalide on 6 main liver microsomal CYP450 isoenzymes was evaluated by using probe drugs; the induction and inhibition activities in vivo of butylphthalide on CYP450 isoenzymes were evaluated by NBP ig dosing (160 mg x kg(-1)) and iv dosing (20 mg x kg(-1)) in rats. After adding the specific inhibitors of CYP2C11, 2E1 and 3A 1/2 for rat, CYP2C19, 2E1 and 3A4/5 for human, the metabolism of NBP in rat and human liver microsomes were reduced 38.8%, 86.2%, 78.4% and 51.0%, 92.0%, 58.9% of control, respectively. The metabolic rates of NBP in CYP2E1 and 3A 1/2 induced rat liver microsomes were increased 25.5% and 68.9%. High concentration of NBP (≥ 200 μmol x L(-1), in vitro) could inhibit the activities of CYP1A2, 2C6, 2C11 and 2D2 in rats, and high concentration of NBP ( ≥ 15 μmol x L(-1), in vitro) could inhibit the activity of CYP2C19 in human. All the results indicated that NBP should be mainly metabolized by CYP2E1, 2C11 and 3A 1/2 in rats and CYP2E1, 2C19 and 3A4/5 in human. High concentration of NBP could inhibit human CYP2C19 in vitro. No significant induction/inhibition effects of NBP were observed on rat liver CYP450 isoforms after ig 160 mg x kg(-1) NBP or iv 20 mg x kg(-1) NBP.

Topics: Animals; Benzofurans; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Humans; Isoenzymes; Liver; Microsomes, Liver; Rats

To explore the effect of 3-n-butylphthalide pretreatment on the delayed neuronal death(DND) and the expreesion of heat shock protein70 (HSP70) in rat hippocampus after ischemia/ reperfusion.. All rats were randomly divided into sham group (n = 36), total cerebral ischemia (TCI) group (n = 36), butylphthalide (NBP) group (n = 6), NBP + TCI group( n = 36), quercetin + NBP + TCI group (n = 6), dimethyl sulfoxide (DMSO) + NBP + TCI group (n = 6). The model of total cerebral ischemia/reperfusion was established by blocking vertebral arteries and carotid arteries. In sham group, TCI group and NBP group, the animals were further divided into instantly, 6 h, 12 h, 1 d, 3 d, 5 d groups according to the time interval after sham operation or TCI. Histological changes of the hippocampus were evaluated using thionin staining under light microscope by determining the delayed neuronal death (DND) and the expression of HSP70 was assayed using immunohistochemistry.. NBP pretreatment could reduce delayed neuronal death in CA1 of hippocampus induced by TCI-reperfusion injury in rats, and up-regulated the expression of HSP70 in CA1 hippocampus of brain ischemic/reperfusion for 5 days. Quercetin blocked the acquirement of the brain ischemic tolerance induced by NBP preconditioning.. 3-n-butylphthalide (NBP) prevents the neurons from ischemia/reperfusion injury through upregulating the expression of HSP70.

Topics: Animals; Benzofurans; CA1 Region, Hippocampal; Cell Death; Cerebral Infarction; HSP70 Heat-Shock Proteins; Ischemic Preconditioning; Neurons; Rats; Rats, Wistar; Reperfusion Injury

Titanium superoxide efficiently catalysed the oxidative esterification of aldehydes with alkylarenes or alcohols, under truly heterogeneous conditions, to afford the corresponding benzyl and alkyl esters in excellent yields. Mechanistic studies have established that this "one pot" direct oxidative esterification process proceeds through a radical pathway, proven by a FTIR spectral study of a titanium superoxide-aldehyde complex as well as spin trapping experiments with TEMPO. The intramolecular version of this protocol has been successfully demonstrated in the concise synthesis of 3-butylphthalide, an anti-convulsant drug.

Topics: Alcohols; Aldehydes; Anticonvulsants; Benzofurans; Benzyl Compounds; Esterification; Oxidants; Oxidation-Reduction; Peroxides; Superoxides; Titanium

To investigate the mechanisms underlying the isomer-selective distribution of 3-n-butylphthalide (NBP) hydroxylated metabolites, 3-hydroxy-NBP (3-OH-NBP) and 10-hydroxy-NBP (10-OH-NBP), across the blood brain barrier (BBB).. After oral administration of NBP (20 mg/kg) to rats, the pharmacokinetics of two major hydroxylated metabolites, 3-OH-NBP and 10-OH-NBP, in plasma and brains were investigated. Plasma and brain protein binding of 3-OH-NBP and 10-OH-NBP was also assessed. To evaluate the influences of major efflux transporters, rats were pretreated with the P-gp inhibitor tariquidar (10 mg/kg, iv) and BCRP inhibitor pantoprazole (40 mg/kg, iv), then received 3-OH-NBP (12 mg/kg, iv) or 10-OH-NBP (3 mg/kg, iv). The metabolic profile of NBP was investigated in rat brain homogenate.. After NBP administration, the plasma exposure of 3-OH-NBP was 4.64 times that of 10-OH-NBP, whereas the brain exposure of 3-OH-NBP was only 11.8% of 10-OH-NBP. In the rat plasma, 60%±5.2% of 10-OH-NBP was unbound to proteins versus only 22%±2.3% of 3-OH-NBP being unbound, whereas in the rat brain, free fractions of 3-OH-NBP and 10-OH-NBP were 100%±9.7% and 49.9%±14.1%, respectively. In the rats pretreated with tariquidar and pantoprazole, the unbound partition coefficient Kp,uu of 3-OH-NBP was significantly increased, while that of 10-OH-NBP showed a slight but not statistically significant increase. Incubation of rat brain homogenate with NBP yielded 3-OH-NBP but not 10-OH-NBP.. The isomer-selective distribution of 10-OH-NBP and 3-OH-NBP across the BBB of rats is mainly attributed to the differences in plasma and brain protein binding and the efflux transport of 3-OH-NBP. The abundant 10-OH-NBP is not generated in rat brains.

Topics: Administration, Oral; Animals; Benzofurans; Biological Transport; Blood-Brain Barrier; Drugs, Chinese Herbal; Isomerism; Male; Rats; Rats, Sprague-Dawley

A series of butylphthalide derivatives (BPDs) 1-8 were isolated from the extract of the dried rhizome of Ligusticum chuanxiong Hort. (Umbelliferae). The cytotoxic activities of BPDs 1-8 were evaluated using a panel of human cancer cell lines. In addition, the SAR analysis and potential anti-invasion activities were investigated. The sp² carbons at C-7 and C-7a appeared to be essential for the cytotoxic activities of BPDs. BPDs 5 and 6 remarkably inhibited the migration and invasion of cancer cells. The anti-invasion activity of dimer 6 was demonstrated to be significantly higher than monomer 5.

Topics: Antineoplastic Agents; Benzofurans; Carcinoma, Hepatocellular; Cell Adhesion; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Humans; Inhibitory Concentration 50; Liver Neoplasms; Molecular Structure; Plant Extracts

1. The aim was to evaluate the prodrug hypothesis by investigating the pharmacokinetics of ZJM-289 and its pharmacological metabolite 3-n-butylphthalide (NBP) in Sprague-Dawley rats and Beagle dogs following intravenous and intragastric administration of ZJM-289. The in vitro metabolic patterns in plasma and microsomal system were assessed to elucidate PK properties. 2. In rats, ZJM-289 was eliminated rapidly (t1/2 = 19.2 ± 3.85 min), along with the fast formation of NBP (formation rate constant ka = 0.29 ± 0.092 min(-1) for intravenous group, and ka = 0.16 ± 0.064 min(-1) for intragastric group), accounting for about 47.4 ± 4.0% of ZJM-289. Both ZJM-289 (t1/2 = 239 ± 70.4 min) and NBP (t1/2 = 249 ± 39.0 min) were depleted slowly in Beagle dogs, with NBP formation rate constant at 0.12 ± 0.052 min(-1) (ka = 0.15 ± 0.040 min(-1) for intragastric group). 3. In rat plasma, ZJM-289 was degraded rapidly (t1/2 = 24.3 ± 0.93 min) at 37 °C, but remained stable with almost no cleavage in dog and human plasma. In hepatic microsomes from rat, dog and human, the hydrolysis metabolites including the active metabolite NBP (M5), and their subsequent hydroxylation and conjugate metabolite, were all detected but varied greatly in the quantities. 4. The findings testified the prodrug design hypothesis that ZJM-289 could be hydrolyzed to NBP. The pharmacokinetic profiles in both rats and dogs brought useful information in the pharmacokinetics prediction in human.

Topics: Administration, Intravenous; Animals; Benzofurans; Chromatography, Liquid; Cinnamates; Dogs; Drug Discovery; Molecular Structure; Nitrates; Nitric Oxide; Prodrugs; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Topics: Adult; Antioxidants; Antipyrine; Benzofurans; Drug Therapy, Combination; Edaravone; Emergency Service, Hospital; Humans; Male; Paraquat

This study investigated the role of L-3-n-Butylphthalide (NBP) in cardiac protection.. The left anterior descending coronary arteries (LAD) of the rats were occluded for 30 min following by 2-h reperfusion to make the ischaemia/reperfusion models. Neonatal cardiomyocytes were cultured and subjected to hypoxia. L-3-n-Butylphthalide was administered intraperitoneally 2 h before the surgery and right after the reperfusion in the in vivo experiments or added to the culture medium in vitro. Haemodynamic parameters were recorded to evaluate the cardiac functions, triphenyltetrazolium chloride (TTC) and Evens blue staining were used to determine the area of risk and infarct area, apoptotic cell numbers were counted with terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining. Western blotting was used to determine the apoptotic protein levels and immune staining to determine the translocation of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein.. Our research showed for the first time that L-3-n-Butylphthalide had great effects in improving cardiac hemodynamic function and decreasing cardiac infarct areas and apoptotic cell numbers in the peri-infarct areas. The apoptotic signals investigation showed that L-3-n-Butylphthalide affected the mitochondrial pathway including Bcl-2 protein expression, inhibition of caspase 3 activation and cytochrome C releasing. Besides, Glyceraldehyde-3-phosphate dehydrogenase protein translocation was inhibited by L-3-n-Butylphthalide treatment, and this effect was mediated by endogenous reactive oxygen species (ROS).. L-3-n-Butylphthalide protects cardiomyocytes from ischaemia/reperfusion-induced apoptosis by antioxidant effect and affecting mitochondrial apoptotic pathway.

Topics: Animals; Antioxidants; Apoptosis; Benzofurans; Blotting, Western; In Situ Nick-End Labeling; Male; Mitochondria; Myocardial Reperfusion Injury; Myocytes, Cardiac; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species

To explore the effects of modulating autophagy on neuroamyloidogenesis in an ischemic stroke model of cultured neuroblastoma 2a (N2a)/Amyloid precursor protein (APP)695 cells.. The ischemic stroke model of N2a/APP695 cells was made by 6h oxygen-glucose deprivation/12h reperfusion (OGDR). Drug administration of 3-methyladenine (3-MA), rapamycin or dl-3-n-butylphthalide (NBP) was started at the beginning of the OGDR and lasted until the end of reperfusion, in order to explore their effects on N2a/APP695 cells under OGDR conditions. Then the cells were incubated in the drug-free and full culture medium under normoxic conditions for 12h. Cell viability and injury were investigated. The key proteins of nuclear factor kappa B (NF-κB) pathway and a key component of autophagy Beclin 1 were detected by Western blotting; immunofluorescence double-staining of amyloid-β (Aβ)1-42 with Beclin 1 was performed to investigate their cellular co-localization relationship; β-secretase and γ-secretase activity assay and Aβ1-42 enzyme-linked immunosorbent assay were performed to investigate the amyloidogenesis.. The results showed that, OGDR enhanced cell injury, autophagy activity, neuroinflammation and Aβ generation in N2a/APP695 cells; down-regulating autophagy by 3-MA and NBP increased cell viability, decreased lactate dehydrogenase (LDH) production, inhibited the activation of NF-κB pathway, suppressed β- and γ-secretase activities and Aβ generation; while up-regulating autophagy by rapamycin got the opposite results; immunofluorescence double-staining results showed elevated Aβ1-42(+) signal was co-localized with Beclin 1(+) signal.. Our data suggested that down-regulating autophagy may inhibit ischemia-induced neuroamyloidogenesis via suppressing the activation of NF-κB pathway. This study might help us to find a new therapeutic strategy to prevent brain ischemic damage and depress the risk of post-stroke dementia.

Topics: Adenine; Amyloid beta-Protein Precursor; Amyloid Precursor Protein Secretases; Animals; Autophagy; Benzofurans; Brain Ischemia; Cell Survival; Mice; Neuroblastoma; Sirolimus; Stroke; Tumor Cells, Cultured

The aim of this study was to explore the effect of butylphthalide on the brain edema, blood-brain barrier of rats of rats after focal cerebral infarction and the expression of Rho A. A total of 195 sprague-dawley male rats were randomly divided into control group, model group, and butylphthalide group (40 mg/kg, once a day, by gavage). The model was made by photochemical method. After surgery 3, 12, 24, 72, and 144 h, brain water content was done to see the effect of butylphthalide for the cerebral edema. Evans blue extravasation method was done to see the changes in blood-brain barrier immunohistochemistry, and Western blot was done to see the expression of Rho A around the infarction. Compared with the control group, the brain water content of model group and butylphthalide group rats was increased, the permeability of blood-brain barrier of model group and butylphthalide group rats was increased, and the Rho A protein of model group and butylphthalide group rats was increased. Compared with the model group, the brain water content of butylphthalide group rats was induced (73.67 ± 0.67 vs 74.14 ± 0.46; 74.89 ± 0.57 vs 75.61 ± 0.52; 77.49 ± 0.34 vs 79.33 ± 0.49; 76.31 ± 0.56 vs 78.01 ± 0.48; 72.36 ± 0.44 vs 73.12 ± 0.73; P < 0.05), the permeability of blood-brain barrier of butylphthalide group rats was induced (319.20 ± 8.11 vs 394.60 ± 6.19; 210.40 ± 9.56 vs 266.40 ± 7.99; 188.00 ± 9.22 vs 232.40 ± 7.89; 288.40 ± 7.86 vs 336.00 ± 6.71; 166.60 ± 6.23 vs 213.60 ± 13.79; P < 0.05), and the Rho A protein of butylphthalide group rats was decreased (western blot result: 1.2230 ± 0.0254 vs 1.3970 ± 0.0276; 1.5985 ± 0.0206 vs 2.0368 ± 0.0179; 1.4229 ± 0.0167 vs 1.7930 ± 0.0158;1.3126 ± 0.0236 vs 1.5471 ± 0.0158; P < 0.05). The butylphthalide could reduce the brain edema, protect the blood-brain barrier, and decrease the expression of Rho A around the infarction.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Brain; Brain Edema; Cerebral Cortex; Cerebral Infarction; Disease Models, Animal; Immunohistochemistry; Male; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; rhoA GTP-Binding Protein; Time Factors; Water

3-n-Butylphthalide (NBP) [(±)-3-butyl-1(3H)-isobenzofuranone] is an anti-cerebral-ischemia drug. Moderate hepatotoxicity has been observed in clinical applications. One of the major metabolites, 3-N-acetylcysteine-NBP, has been detected in human urine, indicating the formation of a reactive metabolite. We elucidated the formation mechanism of the reactive metabolite and its association with the hepatotoxicity of NBP. The in vitro incubations revealed that 3-glutathione-NBP (3-GSH-NBP) was observed only in fresh rat liver homogenate rather than in liver microsomes, liver cytosol, or liver 9,000g supernatant supplemented with NADPH and GSH. We also detected 3-GSH-NBP when 3'-phosphoadenosine-5'-phosphosulfate was added in GSH-fortified human liver cytosol (HLC). The formation of 3-GSH-NBP was 39.3-fold higher using 3-hydroxy-NBP (3-OH-NBP) as the substrate than NBP. The sulfotransferase (SULT) inhibitors DCNP (2,6-dichloro-4-nitrophenol) and quercetin suppressed 3-GSH-NBP formation in HLC by 75 and 82%, respectively, suggesting that 3-OH-NBP sulfation was involved in 3-GSH-NBP formation. Further SULT phenotyping revealed that SULT1A1 is the major isoform responsible for the sulfation. Dose-dependent toxicity was observed in primary rat hepatocytes exposed to 3-OH-NBP, with an IC50 of approximately 168 μM. Addition of DCNP and quercetin significantly increased cell viability, whereas l-buthionine-sulfoximine (a GSH depleter) decreased cell viability. Overall, our study revealed the underlying mechanism for the bioactivation of NBP is as follows. NBP is first oxidized to 3-OH-NBP and further undergoes sulfation to form 3-OH-NBP sulfate. The sulfate spontaneously cleaves off, generating highly reactive electrophilic cations, which can bind either to GSH to detoxify or to hepatocellular proteins to cause undesirable side effects.

Topics: Acetylcysteine; Animals; Arylsulfotransferase; Benzofurans; Biotransformation; Cell Survival; Cells, Cultured; Cytosol; Dose-Response Relationship, Drug; Glutathione; Hepatocytes; Humans; Hydroxylation; Kidney; Liver; Male; Microsomes, Liver; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Sulfur Compounds

The herbal extract 3-n-butylphthalide (NBP) is used in clinical practice for ischemic patients in China. It has been shown to have various neuroprotective effects both in vitro and in vivo. In the present study, the effects of NBP on learning and memory decline in the senescence-accelerated mouse prone-8 (SAMP8) animal model were investigated. Intragastric NBP administration to 4-month-old SAMP8 mice for 2 months significantly improved spatial learning and memory ability. Moreover, the loss of choline acetyltransferase (ChAT)-positive neurons in the medial septal nucleus and the vertical limb of the diagonal band in SAMP8 mice was slowed down, as was the decline in the protein and mRNA expression of ChAT in the hippocampus, cerebral cortex, and forebrain. These results demonstrated that NBP treatment starting at the age of 4 months protected from the learning/memory deficits with aging of SAMP8 mice, and that this effect might be mediated by preventing the decline of the central cholinergic system.

Topics: Aging; Animals; Benzofurans; Cerebral Cortex; Choline O-Acetyltransferase; Cognition; Hippocampus; Male; Maze Learning; Mice; Prosencephalon; Septal Nuclei

In the present study, a series of hydrogen sulfide (H2S) releasing derivatives (8a–g and 9a–f) of 3-n-butylphthalide (NBP) were designed, synthesized and biologically evaluated. The most promising compound 8e significantly inhibited the adenosine diphosphate (ADP) and arachidonic acid (AA)-induced platelet aggregation in vitro, superior to NBP, ticlopidine hydrochloride and aspirin. Furthermore, 8e could slowly produce moderate levels of H2S in vitro, which could be beneficial for improving cardiovascular and cerebral circulation. Most importantly, 8e protected against the collagen and adrenaline induced thrombosis in mice, and exhibited greater antithrombotic activity than NBP and aspirin in rats. Overall, 8e could warrant further investigation for the treatment of thrombosis-related ischemic stroke.

Topics: Adenosine Diphosphate; Administration, Oral; Animals; Benzofurans; Drug Design; Fibrinolytic Agents; Hydrogen Sulfide; Inhibitory Concentration 50; Male; Mice, Inbred ICR; Platelet Aggregation; Platelet Aggregation Inhibitors; Rabbits; Rats, Sprague-Dawley; Survival Analysis

Cognitive impairment is a common finding in patients with chronic obstructive pulmonary disease (COPD), but little attention has been focused on therapeutic intervention for this complication. Chronic intermittent hypoxia hypercapnia (CIHH) exposure is considered to be responsible for the pathogenesis of COPD. Dl-3n-Butylphthalide (NBP), extracted from Apium graveolens Linn, has displayed a broad spectrum of neuroprotective properties. Our study aimed to investigate the potential of NBP on CIHH-induced cognitive deficits. The cognitive function of rats after CIHH exposure was evaluated by the Morris water maze, which showed that the NBP treated group performed better in the navigation test. NBP activated BDNF and phosphorylated CREB, the both are responsible for neuroprotection. Additionally, NBP decreased CIHH induced apoptosis. Moreover, NBP further induced the expression of HIF-1α, accompanied by the up-regulation of the autophagy proteins Bnip3, Beclin-1 and LC3-II. Finally, NBP also reversed the decreased expression of SIRT1 and PGC-1α, but the expression of Tfam, Cox II and mtDNA remained unchanged. These results suggested that the neuroprotective effects of NBP under CIHH condition possibly occurred through the inhibition of apoptosis, promotion of hypoxia-induced autophagy, and activation of the SIRT1/PGC-1α signalling pathway, while stimulation of mitochondrial biogenesis may not be a characteristic response.

Topics: Animals; Apoptosis; Benzofurans; Brain-Derived Neurotrophic Factor; Drug Evaluation, Preclinical; Hypercapnia; Hypoxia, Brain; Male; Maze Learning; Memory; Mitochondrial Turnover; Neuroprotective Agents; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; Pulmonary Disease, Chronic Obstructive; Rats; Rats, Sprague-Dawley; Sirtuin 1; Transcription Factors

Alzheimer's disease (AD) is a progressive neurodegenerative disease. Amyloid-β protein (Aβ), the hallmark of AD, invokes a cascade of mitochondrial dysfunction and eventually leads to neuronal death. l-3-n-Butylphthalide (l-NBP) has shown the potent neuroprotective effects in stroke and AD animal models. The present study is to evaluate the neuroprotective effect of l-NBP on Aβ25-35-induced neuronal injury and the possible mechanism in the human neuroblastoma SH-SY5Y cells. Our results showed that l-NBP significantly attenuated Aβ25-35-induced cell death and reduced neuronal apoptosis. l-NBP significantly inhibited Aβ25-35-induced mitochondrial dysfunction, including mitochondrial membrane potential reduction, and reactive oxygen species production. Furthermore, l-NBP could partially reverse the elevations of Aβ25-35-induced active caspase-3, caspase-9, and cytochrome c expressions, and the downregulation of anti-apoptosis protein Bcl-2. Moreover, l-NBP markedly inhibited the activations of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase/stress-activated protein kinase signaling pathway. These results demonstrated that l-NBP was capable of protecting neuronal cells from Aβ25-35-induced toxicity through a mitochondrial-dependent apoptotic pathway. Thus, l-NBP shows promising candidate of multi-target neuronal protective agent for the treatment of AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Apoptosis; Benzofurans; Caspase 3; Caspase 9; Humans; MAP Kinase Signaling System; Membrane Potential, Mitochondrial; Mitochondria; Molecular Structure; Neuroblastoma; Neuroprotective Agents; p38 Mitogen-Activated Protein Kinases; Peptide Fragments; Reactive Oxygen Species; Signal Transduction

3-n-Butylphthalide (NBP) is a cardiovascular drug widely used in China for the treatment of cerebral ischemic stroke. Our previous study showed that NBP underwent extensive metabolism in humans and that 10-keto-NBP (M2), 3-hydroxy-NBP (M3-1), 10-hydroxy-NBP (M3-2) and NBP-11-oic acid (M5-2) were the major circulating metabolites. A better understanding of the plasma exposures of NBP and its four major metabolites is crucial to supporting the safety evaluation, good clinic practice and discovery of new antistroke drugs. Herein, a liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of NBP, M2, M3-1, M3-2, and M5-2 in human plasma. To improve assay sensitivity and achieve simultaneous analysis, M3-1 and M5-2 were monitored in (-)ESI (electrospray) mode within the first 3.5 min and NBP, M2, and M3-2 thereafter in (+)ESI mode. Deuterated internal standards for all analytes were synthesized to compensate for the impact of matrix effects. Based on the vast differences in physicochemical properties among the five analytes and the necessary baseline separation of two isomers (i.e., M3-1 and M3-2), gradient elution comprising a mobile phase of methanol-acetonitrile-5 mM ammonium acetate was used after methanol-induced protein precipitation of plasma samples. The developed method was linear in the concentration range of 3.00-800 ng/ml for NBP and M2, and 3.00-2400 ng/ml for M3-1, M3-2, and M5-2. The lower limit of quantitation was 3.00 ng/ml for each analyte. The intra- and inter-day accuracy and precision were within acceptable limits (±15%) at all concentrations. The method was successfully applied to characterize the pharmacokinetic profiles of NBP and its major metabolites following a single oral administration of 200mg NBP to healthy volunteers.

Topics: Benzofurans; Chromatography, Liquid; Deuterium; Humans; Reference Standards; Tandem Mass Spectrometry

In search of novel anti-ischemic stroke agents with higher potency than a known drug 3-n-butylphthalide (NBP), a series of hybrids ((S)- and (R)-5a-f) from optically active ring-opened NBP derivative and isosorbide were synthesized for evaluating their anti-ischemic stroke activity. Compound (S)-5e displayed the strongest activity in inhibiting the adenosine diphosphate (ADP) and arachidonic acid (AA)-induced platelet aggregation in vitro, with 10.0- and 8.4-fold more effectiveness than (S)-NBP, respectively. Furthermore, (S)-5e was stable in artificial gastrointestinal fluids and could penetrate the blood-brain barrier (BBB) with an appreciate lipid/water partition coefficient relative to (S)-NBP. More importantly, oral treatment with (S)-5e protected from acute thrombosis and inhibited the ischemia/reperfusion-related brain injury in animals. Our findings suggest that (S)-5e may be promising for further evaluation for the intervention of ischemic stroke.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Isosorbide; Platelet Aggregation Inhibitors; Rats; Stereoisomerism; Stroke

This study investigated the effects of different celery (Apium graveolens) seed extracts on blood pressure (BP) in normotensive and deoxycorticosterone acetate-induced hypertensive rats. The hexanic, methanolic, and aqueous-ethanolic extracts were administered intraperitoneally and their effects on BP and heart rate (HR) were evaluated in comparison with spirnolactone as a diuretic and positive control. Also, the amount of n-butylphthalide (NBP), as an antihypertensive constituent, in each extract was determined by HPLC. The results indicated that all extracts decreased BP and increased the HR in hypertensive rats, but had no effect on normotensive rats. The data showed that administration of 300 mg/kg of hexanic, methanolic, and aqueous-ethanolic (20/80, v/v) extracts of the celery seed caused 38, 24, and 23 mmHg reduction in BP and 60, 25, and 27 beats per minute increase in the HR, respectively. Also, the HPLC analysis data revealed that the content of NBP in the hexanic extract was 3.7 and 4 times greater than methanolic and aqueous-ethanolic extracts. It can be concluded that celery seed extracts have antihypertensive properties, which appears to be attributable to the actions of its active hydrophobic constitutes such as NBP and can be considered as an antihypertensive agent in chronic treatment of elevated BP.

Topics: Animals; Antihypertensive Agents; Apium; Benzofurans; Blood Pressure; Heart Rate; Humans; Hypertension; Male; Plant Extracts; Rats; Rats, Wistar; Seeds

Activation of astrocytes is a common feature of Alzheimer's disease (AD). Proinflammatory molecules produced by activated astrocytes contribute to neuronal damage in AD. Moreover, dl-3-n-butylphthalide (NBP) has been reported to attenuate astroglial activation and exert neuroprotective effects in AD transgenic mice. However, the mechanism by which NBP inhibits activated astrocytes is poorly understood.. In this study, the primary astrocytes were obtained from the cerebral cortices of 1-day-old Sprague-Dawley rats. The levels of GFAP, COX-2, NF-κB, and IκBα were examined by Western blotting and the levels of TNF-α and IL-6 were determined by ELISA.. NBP inhibited the amyloid-β (Aβ)-induced activation of astrocytes and the up-regulation of proinflammatory molecules. Importantly, NBP markedly suppressed Aβ-induced IκBα degradation and nuclear factor-κB (NF-κB) translocation.. Our results suggest that NBP attenuates Aβ-induced activation of astrocytes and neuroinflammation via inhibition of the NF-κB signaling pathway.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Astrocytes; Benzofurans; Cells, Cultured; Cerebral Cortex; Cyclooxygenase 2; I-kappa B Proteins; Interleukin-6; Mice; Mice, Transgenic; Nerve Tissue Proteins; Neuroprotective Agents; NF-kappa B; NF-KappaB Inhibitor alpha; Rats; Rats, Sprague-Dawley; Signal Transduction; Transcription Factor RelA; Tumor Necrosis Factor-alpha; Up-Regulation

Topics: Benzofurans; China; Fibrinolytic Agents; Humans; Stroke

A novel and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to determine the exposure of 3-n-butylphthalide (NBP) enantiomers in human plasma. The NBP enantiomers were extracted from human plasma using methyl tert-butyl ether. The baseline separation of R-(+)-NBP and S-(-)-NBP was achieved within 11.0min using a teicoplanin-based Astec Chirobiotic T column (250mm×4.6mm i.d., 5μm) under isocratic conditions at a flow rate of 0.6mL/min. The selection of the chiral stationary phase and the effect of the mobile phase composition on the resolution of the enantiomers were discussed. The selectivity, linearity, precision, accuracy, matrix effect, recovery, and stability were evaluated under optimized conditions. The LC-MS/MS method using 200μL of human plasma was linear over the concentration range of 5.00-400ng/mL for each enantiomer. The lower limit of quantification (LLOQ) for both enantiomers was 5.00ng/mL. The intra- and inter-assay precision values of the replicated quality control samples were within 8.0% for each enantiomer. The mean accuracy values for the quality control samples were within ±6.1% of the nominal values for R-(+)-NBP and S-(-)-NBP. No chiral inversion was observed during sample storage, preparation, and analysis. The method proved suitable for enantioselective pharmacokinetic studies of NBP after an oral administration of a therapeutic dose of racemic NBP.

Topics: Benzofurans; Chromatography, Liquid; Drug Stability; Humans; Least-Squares Analysis; Male; Reproducibility of Results; Sensitivity and Specificity; Stereoisomerism; Tandem Mass Spectrometry; Teicoplanin

To elucidate the expression and identify the functional changes of 2 pore domain potassium channel TREK-1 during cardiac hypertrophy in rats, left ventricular hypertrophy was induced by subcutaneous injection with isoproterenol. Western blot was used to detect the expression of TREK-1 channel protein, and inside-out and whole-cell recordings were used to record TREK-1 currents. The results showed that TREK-1 protein expression in endocardium was slightly higher than that in epicardium in control left ventricles. However, it was obviously upregulated by 89.8% during hypertrophy, 2.3-fold higher than in epicardium. Mechanical stretch, intracellular acidification, and arachidonic acid could activate a TREK-1-like current in cardiomyocytes. The slope conductances of cardiac TREK-1 and CHO/TREK-1 channels were 123 ± 7 and 113 ± 17 pS, respectively. The TREK-1 inhibitor L-3-n-butylphthalide (10 μM) reduced the currents in CHO/TREK-1 cells, normal cardiomyocytes, and hypertrophic cardiomyocytes by 48.5%, 54.3%, and 55.5%, respectively. The percentage of L-3-n-butylphthalide-inhibited outward whole-cell current in hypertrophic cardiomyocytes (23.7%) was larger than that in normal cardiomyocytes (14.2%). The percentage of chloroform-activated outward whole-cell current in hypertrophic cardiomyocytes (58.3%) was also larger than normal control (40.2%). Our results demonstrated that in hypertrophic rats, TREK-1 protein expression in endocardium was specifically increased and the ratio of TREK-1 channel current in cardiac outward currents was also enhanced. TREK-1 might balance potassium ion flow during hypertrophy and might be a potential drug target for heart protection.

Topics: Animals; Benzofurans; Blotting, Western; CHO Cells; Cricetinae; Cricetulus; Disease Models, Animal; Gene Expression Regulation; Heart Ventricles; Hypertrophy, Left Ventricular; Isoproterenol; Male; Myocytes, Cardiac; Patch-Clamp Techniques; Potassium Channels, Tandem Pore Domain; Rats; Rats, Sprague-Dawley

In the absence of a cure for Parkinson's disease, development of preventive medications for this devastating disease is particularly encouraged. Dl-3-n-butylphthalide (NBP), an established natural antioxidant for clinical stroke treatment in China, can reportedly reduce beta-amyloid-induced neuronal toxicity in cultured neuronal cells, and attenuate neurodegenerative changes in aged rats. However, whether or not NBP confers neuroprotection in parkinsonian models is still unknown. In this study, we investigated the effects of NBP in rotenone models for Parkinson's diseases. In a cellular model, pretreatment with NBP enhanced cell viability by decreasing nuclear fragmentation, retaining mitochondrial membrane potential, and preventing reactive oxygen species (ROS) from generation. In a rodent model, 2-week treatment with NBP was able to ameliorate apomorphine-evoked rotations by 48% and rescue dopaminergic (DA) neurons by 30% and striatal DA terminal by 49%. Furthermore, NBP upregulated the vesicular monoamine transporter 2 gene expression in vitro and in vivo. Together, NBP protects DA neurons likely by reducing oxidative stress, offering an alternative neuroprotective medication for Parkinson's disease.

Topics: Animals; Antioxidants; Antiparkinson Agents; Benzofurans; Cell Line; Cell Survival; Disease Models, Animal; Dopaminergic Neurons; Humans; Neuroprotective Agents; Parkinsonian Disorders; Rats; Rotenone; Treatment Outcome

Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative disease characterized by progressive muscular atrophy, paralysis and bulbar symptoms. Transgenic mice over-expressing human mutant Cu/Zn superoxide dismutase-1 (SOD1) mimicked the pathological phenotype of ALS. dl-3-n-butylphthalide (dl-NBP) has been demonstrated to play a neuroprotective role in cerebral ischemia, vascular dementia, and Alzheimer's disease. In the current study, we examined the effect of dl-NBP in Tg (SOD1-G93A) transgenic mice, a well-studied model of ALS. Following the symptomatic onset of disease, oral administration of dl-NBP significantly improved motor performance, extended the survival interval, attenuated motor neuron loss, and delayed motor unit reduction compared to vehicle controls. These observations were further corroborated by the significant reduction in immunoreactivity of CD11b and glial fibrillary acidic protein (GFAP), markers for microglia and astrocytes, respectively. Additionally, downregulation of nuclear factor κB (NF-κB) p65 and tumor necrosis factor-α (TNF-α) protein levels and a slight upregulation of NF-E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were found in the spinal cord of Tg (SOD1-G93A) mice treated by dl-NBP. These results suggest that dl-NBP might be a promising compound in the treatment of ALS. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.

Topics: Amyotrophic Lateral Sclerosis; Animals; Benzofurans; Disease Models, Animal; Mice; Mice, Transgenic; Motor Neurons; Neuroglia; Neuroprotective Agents; Spinal Cord; Superoxide Dismutase

Pharmacological compounds that release nitric oxide (NO) have been recognized as the potential therapeutic agents for acute stroke. (S)-ZJM-289 is a novel NO-releasing derivative of 3-n-butylphthalide (NBP) with enhanced anti-platelet and anti-thrombotic actions. The present study was performed to investigate the neuroprotective effects and related mechanisms of (S)-ZJM-289 on ischemic neuronal injury in vitro and in vivo. Primary cortical neuronal cultures were exposured to oxygen-glucose deprivation followed by recovery (OGD/R), a model of ischemia-like injury, and treated with (S)-ZJM-289 before OGD. In vitro results showed that (S)-ZJM-289 attenuated OGD/R-induced neuronal injury, which was associated with the maintenance of mitochondrial integrity and function by alleviating intracellular calcium overload and reactive oxygen species (ROS) accumulation, preventing mitochondrial membrane depolarization and preserving respiratory chain complexes activities. Moreover, (S)-ZJM-289 treatment suppressed mitochondrial release of cytochrome c (cyt c) and nuclear translocation of apoptosis-inducing factor (AIF), thereby blocking mitochondria-mediated cell death, which may be partially mediated by up-regulation of Hsp70. The neuroprotection by (S)-ZJM-289 was also studied using a model of middle cerebral artery occlusion (MCAO). Oral administration of (S)-ZJM-289 at the onset of reperfusion for 3d significantly reduced the brain infarct size, improved neurological deficit and prevented neuronal loss and apoptosis. In current study, (S)-ZJM-289 appears to be more potent in ischemic neuroprotection than NBP, in particular at the lower doses, which may be due to the synergistic action of NBP and NO. These findings point to that (S)-ZJM-289 could be an attractive alternative to NBP in preventing the process of ischemia/reperfusion (I/R) injury.

Topics: Animals; Benzofurans; Cell Death; Cinnamates; Disease Models, Animal; Hypoxia-Ischemia, Brain; Male; Mitochondrial Diseases; Nerve Degeneration; Neuroprotective Agents; Nitrates; Primary Cell Culture; Rats; Rats, Sprague-Dawley; Reperfusion Injury

L-3-n-butylphthalide (L-NBP), an extract from seeds of Apium graveolens Linn (Chinese celery), has been shown to have neuroprotective effects on cerebral ischemic, vascular dementia and amyloid-β (Aβ)-induced animal models by inhibiting oxidative injury, neuronal apoptosis and glial activation, regulating amyloid-β protein precursor (AβPP) processing and reducing Aβ generation. The aim of the present study was to examine the effect of L-NBP on learning and memory in AβPP and presenilin 1 (PS1) double-transgenic AD mouse model (AβPP/PS1) and the mechanisms of L-NBP in reducing Aβ accumulation and tau phosphorylation. Twelve-month old AβPP/PS1 mice were given 15 mg/kg L-NBP by oral gavage for 3 months. L-NBP treatment significantly improved the spatial learning and memory deficits compared to the vehicle-treated AβPP/PS1 mice, whereas L-NBP treatment had no effect on cerebral Aβ plaque deposition and Aβ levels in brain homogenates. However, we found an L-NBP-induced reduction of tau hyperphosphorylation at Ser199, Thr205, Ser396, and Ser404 sites in AβPP/PS1 mice. Additionally, the expressions of cyclin-dependent kinase and glycogen synthase kinase 3β, the most important kinases involved in tau phosphorylation, were markedly decreased by L-NBP treatment. The effects of L-NBP on decreasing tau phosphorylation and kinases activations were further confirmed in neuroblastoma SK-N-SH cells overexpressing wild-type human AβPP695 (SK-N-SH AβPPwt). L-NBP shows promising candidate of multi-target neuronal protective agent for the treatment of Alzheimer's disease.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Amyloid beta-Protein Precursor; Analysis of Variance; Animals; Benzofurans; Brain; Cell Line, Tumor; Cognition Disorders; Cyclin-Dependent Kinase 5; Disease Models, Animal; Gene Expression Regulation; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Male; Maze Learning; Mice; Mice, Transgenic; Neuroblastoma; Neuroprotective Agents; Phosphorylation; Presenilin-1; Reaction Time; Spatial Behavior; tau Proteins; Time Factors

3-n-Butylphthalide (NBP) is a compound extracted from Chinese celery and is used as an anti-hypertensive herbal medicine for treating stroke patients. The aim of this study is to demonstrate the effects and mechanisms of this compound through in vitro and in vivo experiments. Culture experiments were performed by adding hydrogen peroxide (H(2)O(2)) to SH-SY5Y cells. From the MTT assay result, enhanced cell survival was observed with DL-NBP treatment, regardless of whether they are added before, simultaneously with or after the addition of H(2)O(2). For the in vivo experiment, Spontaneously Hypertensive rats and Wistar Kyoto control rats with chronic cerebral ischemia, which were induced by bilateral transection of the common carotid arteries, were given DL-NBP. Their performances in the place navigation test and spatial probe test in the Morris Water Maze have significantly improved compared with the DL-NBP untreated animals, indicating an improvement in spatial learning and memory in the ischemic-animals. In addition, in the chick embryonic chorioallantoic membrane assay, angiogenesis was more vigorous under the effects of DL-NBP, together with increased expression of growth factors, VEGF, VEGF-receptor and bFGF. All these suggested that one of the mechanisms of DL-NBP might be ameliorating vascular dementia and promoting angiogenesis.

Topics: Animals; Benzofurans; Cell Line, Tumor; Chick Embryo; Dementia, Vascular; Drugs, Chinese Herbal; Humans; Immunohistochemistry; Male; Maze Learning; Neovascularization, Physiologic; Neuroprotective Agents; Rats; Rats, Inbred SHR; Rats, Inbred WKY

We have previously demonstrated that dl-3n-butylphthalide (NBP) has a potential angiogenic activity. In this study, we investigated the angiogenic effect of NBP and the molecular mechanisms underlying NBP-mediated angiogenesis. Zebrafish embryos and human umbilical vein endothelial cells were treated with various doses of NBP and several signaling pathway inhibitors. NBP induced ectopic subintestinal vessel production in zebrafish embryos and induced invasion, migration, and endothelial cell tube formation of human umbilical vein endothelial cells in a dose-dependent manner. These NBP-induced angiogenic effects were partially suppressed by SU5402, a fibroblast growth factor receptor 1 inhibitor; U0126, an extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor; LY294002, a phosphatidylinositol 3-kinase inhibitor; 1L6-hydroxymethyl-chiro-inositol-2-(R)-2-O-methyl-3-O-octadecyl-sn-glycerocarbonate, an Akt inhibitor; cavtratin, an endothelial nitric oxide synthase (eNOS) inhibitor and completely inhibited by a combination of U0126 and LY294002. NBP enhanced phosphorylation of ERK1/2 and fibroblast growth factor receptor 2 expression, which were inhibited by U0126. NBP increased the phosphorylation of Akt and eNOS at serine 1177, which was blocked by LY294002. NBP-stimulated nitric oxide production, which was reduced by LY294002. Our data demonstrated that (1) NBP promoted angiogenesis and (2) the angiogenic effects of NBP were mediated by the ERK1/2 and phosphatidylinositol 3-kinase/Akt-eNOS signaling pathways. Our findings suggest that NBP could be a novel agent for therapeutic angiogenesis in ischemic diseases.

Topics: Animals; Benzofurans; Dose-Response Relationship, Drug; Human Umbilical Vein Endothelial Cells; Humans; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neovascularization, Physiologic; Neuroprotective Agents; Nitric Oxide Synthase Type III; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction; Zebrafish

3-n-Butylphthalide (NBP) has been shown to have protective effects against ischemic stroke. In the present study, we investigated effects of l-3-n-butylphthalide (l-NBP) on the learning and memory impairment induced by chronic cerebral ischemia in rats. Male Wistar rats were administered 20 mg/kg l-NBP by gavage daily for 30 days after the bilateral common carotid artery clamping (two-vessel occlusion, 2-VO). Results showed that daily treatments of 20 mg/kg l-NBP significantly attenuated spatial learning deficits in Morris water maze (MWM) task. Results of long-term potentiation (LTP) indicated that treatment with 20 mg/kg l-NBP attenuated the inhibition of LTP in rat model of 2-VO. Moreover, l-NBP reduced glial fibrillary acidic protein (GFAP)-positive astrocytes induced by chronic cerebral ischemia. The present findings demonstrate the protective effect of l-NBP on chronic cerebral ischemia-induced hippocampus injury, which supports using l-NBP for therapy of cerebral ischemia in the future.

Topics: Animals; Benzofurans; Brain Ischemia; Chronic Disease; Cognition Disorders; Male; Maze Learning; Neuroprotective Agents; Random Allocation; Rats; Rats, Wistar

The objective of the present study was to investigate the effects of 3-n-butylphthalide (NBP) on a 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of Parkinson's disease (PD) and to illustrate the potential mechanism of autophagy in this process. For this purpose, rat PC12 pheochromocytoma cells were treated with MPP+ (1 mM) for 24 h following pretreatment with NBP (0.1 mM). Cell metabolic viability was determined by the MTT assay and cell ultrastructure was examined by transmission electron microscopy. The intracellular distribution and expression of α-synuclein and microtubule-associated protein light chain 3 (LC3) were detected by immunocytochemistry and Western blotting. Our results demonstrated that: 1) NBP prevented MPP+-induced cytotoxicity in PC12 cells by promoting metabolic viability. 2) NBP induced the accumulation of autophagosomes in MPP+-treated PC12 cells. 3) Further study of the molecular mechanism demonstrated that NBP enhanced the colocalization of α-synuclein and LC3 and up-regulated the protein level of LC3-II. These results demonstrate that NBP protects PC12 cells against MPP+-induced neurotoxicity by activating autophagy-mediated α-synuclein degradation, implying that it may be a potential effective therapeutic agent for the treatment of PD.

Topics: 1-Methyl-4-phenylpyridinium; Animals; Apium; Autophagy; Benzofurans; Blotting, Western; Cell Survival; Immunohistochemistry; Microscopy, Electron, Transmission; Neuroprotective Agents; Parkinson Disease, Secondary; PC12 Cells; Rats; Seeds

Topics: Alcohol-Related Disorders; Animals; Benzofurans; Glutamic Acid; Hippocampus; Male; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate

Bone marrow stem cells (BMSCs) have been one of the most important cell sources for cell replacement therapy (CRT) in cerebral infarction. However, long-lasting oxidative stress during stroke, which plays an important role in neuron damage, deteriorates the microenvironment for cell survival, differentiation and removal. Thus the outcome of CRT in ischemic diseases was poor. DL-3-n-Butylphthalide (NBP) has protective effects on ischemic brain tissue through multiple mechanisms and has been used for stroke treatment in China for several years. In this study, hydrogen peroxide (H(2)O(2)) was used to induce oxidative stress injury to rat bone marrow stem cells (rBMSCs), imitating the microenvironment surrounding transplanted cells in the ischemic brain in vitro. The protective effects of NBP on rBMSCs against apoptosis induced by oxidative stress were investigated. Our results indicated that NBP could protect rBMSCs against apoptosis due to antioxidative properties and modulation of PI3K/Akt pathway. NBP could be used in combination with BMSCs for the treatment of cerebral infarction by improving the oxidative stress microenvironments and cell survival, however, further studies remain warranted.

Topics: Animals; Antioxidants; Apoptosis; Benzofurans; Bone Marrow Cells; Enzyme Activation; Hematopoietic Stem Cells; Hydrogen Peroxide; Oxidants; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Signal Transduction

DL-3-n-Butylphthalide (NBP) is a synthetic compound based on L-3-n-Butylphthalide which was isolated from seeds of Apium graveolens. The present study aims at evaluating the outcome of NBP given prior to and after the onset of ischemic stroke in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Stroke was induced by the middle cerebral artery occlusion (MCAO) in SHR and WKY. For pre-treatment, NBP was administered to SHR and WKY daily for two months prior to MCAO. For post-treatment, NBP was given daily for seven consecutive days after MCAO. Seven days post-surgery, rats were tested for the presence of neurological deficits. Magnetic resonance imaging (MRI) and 2,3,5-triphenyltetrazolium chloride (TTC) staining were employed to calculate the infarct volume. The cerebral cortex and corpus striatum in the ischemic penumbra area were examined microscopically for pathological changes. In SHR, NBP pre- and post-treatment significantly lowered neurological deficit scores, reduced infarct volume, and minimized pathological changes in the penumbra area when compared to oil-vehicle treated controls. In WKY, these beneficial effects were observed only in the post-treatment group. The beneficial effects of NBP post-treatment were greater in WKY than in SHR. Results indicated that NBP could exert both preventive and therapeutic effects on ischemic stroke in SHR, but only exerted therapeutic effect in WKY.

Topics: Analysis of Variance; Animals; Antioxidants; Benzofurans; Brain Infarction; Brain Injuries; Cerebral Cortex; Disease Models, Animal; Infarction, Middle Cerebral Artery; Magnetic Resonance Imaging; Male; Nervous System Diseases; Neurologic Examination; Rats; Rats, Inbred SHR; Rats, Inbred WKY; Tetrazolium Salts

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by progressive death of the upper and lower motor neurons. Transgenic mice over-expressing a mutant form of the human SOD1 gene develop an ALS-like phenotype. Currently, there is no effective treatment or drug for the fatal disease. Previous studies reported potent efficacy of dl-3-n-butylphthalide (DL-NBP) for several neurodegenerative disorders and cerebral ischemia. SOD1-G93A mice are a mouse model of ALS. In this study, we investigated the efficacy of DL-NBP on this ALS mouse model.. Sixty SOD1-G93A female mice were divided into four groups. The vehicle control group received 0 mg×kg(-1)×d(-1) DL-NBP. The experimental groups received DL-NBP with doses of 30, 60 or 120 mg×kg(-1)×d(-1), respectively. For measurement of motor activity, the hanging wire test and rotarod test were performed. Survival statistics were analyzed by Kaplan-Meier survival curves. The body weight of each mouse was recorded twice per week. The statistical motor unit number estimation (MUNE) technique was used to estimate the number of functioning motor units in gastrocnemius muscle. Muscle morphology was evaluated by hematoxylin and eosin staining. Motor neuron quantitation was performed by Nissl staining and microglia activation was observed by immunohistochemistry.. Oral administration of 60 mg×kg(-1)×d(-1)1 DL-NBP significantly prolonged survival ((164.78 ± 16.67) days) of SOD1-G93A mice compared with vehicle control ((140.00 ± 16.89) days). Treating mice with DL-NBP (60 mg×kg(-1)×d(-1)) significantly decreased the progression rate of motor deficits and suppressed body weight reduction. Furthermore, we found that treating SOD1-G93A mice with DL-NBP (60 mg×kg(-1)×d(-1)) slowed the rate of MUNE reduction (P < 0.01). Motor neurons were remarkably preserved in the anterior horns in mice treated with DL-NBP (60 mg×kg(-1)×d(-1)) at the stage of 19 weeks (P < 0.01). Treating mice with DL-NBP (60 mg×kg(-1)×d(-1)) significantly reduced CD11b immunoreactivity compared with vehicle control mice (P < 0.05). No significant effect was observed in mice treated with DL-NBP of 30 or 120 mg×kg(-1)×d(-1).. The post-disease-onset administration of DL-NBP significantly prolonged survival and improved motor performance in SOD1-G93A mice. DL-NBP may be a potential therapeutic agent for ALS.

Topics: Administration, Oral; Amyotrophic Lateral Sclerosis; Animals; Benzofurans; Disease Models, Animal; Female; Immunohistochemistry; Mice; Mice, Transgenic

To investigate the therapeutic effect of DL-3-n-butylphthalide (DL-NBP) in rats with chronic cerebral hypoperfusion.. Chronic cerebral hypoperfusion was modelled by bilateral permanent occlusion of common carotid arteries in Wistar rats. The therapeutic effect of DL-NBP in hypoperfused rats was evaluated using the Morris water maze task. The levels and deposition of matrix metalloproteinase (MMP) and the amyloid precursor protein β-amyloid 40 (Aβ40) were measured by Western blot analysis and immunohistochemistry in the cerebral cortex and hippocampus.. Treatment with DL-NBP significantly improved the learning and memory ability of hypoperfused rats. Western blot analysis indicated that, in comparison with the sham-operated control group, protein levels of Aβ40 and MMP-2 were significantly increased in the cerebral cortex of hypoperfused rats, and treatment with DL-NBP prevented this hypoperfusion-induced increase in Aβ40 and MMP-2. Immunohistochemical analysis showed that Aβ40 and MMP-2 were deposited in venous endothelial cells at day 3 and in arterial endothelial cells at day 14 after hypoperfusion.. This study indicated that DL-NBP has therapeutic effects on chronic cerebral hypoperfusion and provided a useful insight into the potential molecular mechanisms underlying the therapeutic effect of DL-NBP in chronic cerebral hypoperfusion.

Topics: Amyloid beta-Protein Precursor; Animals; Benzofurans; Blotting, Western; Brain Ischemia; Chronic Disease; Down-Regulation; Immunohistochemistry; Male; Matrix Metalloproteinase 2; Maze Learning; Rats; Rats, Wistar

Topics: Benzofurans; Humans; Neuroprotective Agents; Oxidative Stress

Amyloid precursor protein (APP) is cleaved by α-secretase, within the amyloid-β (Aβ) sequence, resulting in the release of a secreted fragment (αAPPs) and precluding Aβ production. We investigated the effects of a promising anti-AD new drug, l-3-n-butylphthalide (L-NBP), on APP processing and Aβ generation in neuroblastoma SK-N-SH cells overexpressing wild-type human APP695. L-NBP significantly increased αAPPs release, and reduced Aβ generation. The steady-state full-length APP levels were unaffected by L-NBP. It suggested that L-NBP regulated APP processing towards to the non-amyloidogenic α-secretase pathway. Protein kinase C (PKC) and mitogen activated protein (MAP) kinase might be involved in L-NBP-induced αAPPs secretion. L-NBP significantly increased PKCα and ɛ activations, lowered PKCγ activation and increased the phosphorylation of p44/p42 MAPK. Furthermore, PKC and MAPK inhibitors partially reduced L-NBP-induced αAPPs secretion. The results suggested alternative pharmacological mechanisms of L-NBP regarding the treatment of Alzheimer's disease (AD).

Topics: Amyloid beta-Protein Precursor; Benzofurans; Cell Line, Tumor; Cell Survival; Drugs, Chinese Herbal; Gene Expression Regulation, Enzymologic; Humans; MAP Kinase Signaling System; Mitogen-Activated Protein Kinases; Peptide Fragments; Protein Kinase C; Protein Processing, Post-Translational; Seeds

Alzheimer's disease (AD) is the most common form of dementia. Oxidative stress is one of the earliest events in the neurological and pathological changes of AD. L-3-n-butyl-phthalide (L-NBP), an anti-cerebral ischemia agent, has been shown a potential in AD treatment. In this study, we investigated the neuroprotective effect of L-NBP on hydrogen peroxide (H₂O₂)-induced apoptosis in human neuroblastoma SK-N-SH cells. H₂O₂ significantly reduced cell viability and increased the number of apoptotic-like cells, indicating that H₂O₂ induced neurotoxicity. In addition, real-time PCR and western blot studies showed that Bcl-2 and Bcl-w expressions were decreased, and Bax expression was increased with H₂O₂ treatment. Moreover, protein kinase C (PKC) α expression was down-regulated after H₂O₂ treatment. All of these phenotypes induced by H₂O₂ were markedly reversed by L-NBP. Pretreatment with L-NBP significantly increased cell viability of H₂O₂-damaged cells, and reduced H₂O₂-induced neuronal apoptosis. L-NBP treatment at dose of 10 μM inhibited H₂O₂-induced down-regulation of Bcl-2, Bcl-w, and PKCα but also attenuated the overexpression of Bax. PKC inhibitor, calphostin C, significantly attenuated the protective effects of L-NBP. Our findings suggest that L-NBP may protect neurons against H₂O₂-induced apoptosis by modulating apoptosis-related genes and activating PKCα pathway.

Topics: Apoptosis; Apoptosis Regulatory Proteins; bcl-2-Associated X Protein; Benzofurans; Cell Line, Tumor; Cell Survival; Gene Expression; Humans; Hydrogen Peroxide; Naphthalenes; Neuroblastoma; Neuroprotective Agents; Protein Kinase C; Protein Kinase C-alpha; Protein Kinase C-epsilon; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-bcl-2; Signal Transduction

To study the effects of 3-n-butylphthalide (NBP) on the TREK-1 channel expressed in Chinese hamster ovary (CHO) cells.. Whole-cell patch-clamp recording was used to record TREK-1 channel currents. The effects of varying doses of l-NBP on TREK-1 currents were also observed. Current-clamp recordings were performed to measure the resting membrane potential in TREK-1-transfected CHO (TREK-1/CHO) and wild-type CHO (Wt/CHO) cells.. l-NBP (0.01-10 μmol/L) showed concentration-dependent inhibition on TREK-1 currents (IC(50)=0.06±0.03 μmol/L), with a maximum current reduction of 70% at a concentration of 10 μmol/L. l-NBP showed a more potent inhibition on TREK-1 current than d-NBP or dl-NBP. This effect was partially reversed upon washout and was not voltage-dependent. l-NBP 10 μmol/L elevated the membrane potential in TREK-1/CHO cells from -55.3 mV to -42.9 mV. However, it had no effect on the membrane potential of Wt/CHO cells.. 1-NBP potently inhibited TREK-1 current and elevated the membrane potential, which may contribute to its neuroprotective activity.

Topics: Animals; Benzofurans; CHO Cells; Cricetinae; Cricetulus; Dose-Response Relationship, Drug; Inhibitory Concentration 50; Membrane Potentials; Neuroprotective Agents; Patch-Clamp Techniques; Potassium Channels, Tandem Pore Domain; Rats; Stereoisomerism

The aim of this study was to investigate the protective effect of dl-3-n-butylphthalide (NBP) on brain damage in streptozotocin (STZ)-induced diabetic mice subjected to cerebral ischemia.. we pretreated diabetic mice with NBP orally for 4 weeks prior and 2 days after transient common carotid artery occlusion (CCAO) operation. Immunohistochemistry and transmission electron microscopy were performed to investigate the neuronal loss, astrocytes activation, amyloid-beta (Abeta) protein expression, and autophagy activation.. The results showed that diabetes increased stroke-induced neuronal loss, astrocytes activation, Abeta generation, and autophagy activity, while NBP administration attenuated these changes. Immunofluorescence double staining of Abeta with autophagosome-specific antibody LC3 showed that most elevated Abeta(+) signal was co-localized with LC3(+) signal.. Our finding suggests that NBP attenuates Abeta generation promoted by diabetes in ischemia might act through inhibiting abnormally activated neuronal autophagy. Therefore, treatment with NBP to modulate autophagy might provide a novel therapeutic strategy for diabetes by preventing ischemic brain damage and depressing the risk of post-stroke dementia.

Topics: Amyloid beta-Peptides; Animals; Autophagy; Benzofurans; Brain Ischemia; Dementia; Diabetes Mellitus, Experimental; Disease Models, Animal; Male; Mice; Mice, Inbred C57BL; Nerve Degeneration; Neuroprotective Agents; Random Allocation; Treatment Outcome

Most ischemic stroke results from brain blood vessel blockage by platelet-mediated thrombus, and anti-platelet therapy has been demonstrated clinical benefits in the treatment of this disease. In the present work, novel nitric oxide (NO)-releasing derivatives of an anti-ischemic stroke drug 3-n-butylphthalide (NBP) were synthesized. Compounds 7a and 7c exhibited more potent anti-platelet activity than NBP and aspirin, and released a moderate amount of NO, which is beneficial in improving cardiovascular and cerebral circulation. These findings provide an alternative approach to the development of drugs more potent than NBP for the intervention of ischemic stroke.

Topics: Adenosine Diphosphate; Animals; Benzoates; Benzofurans; Blood Platelets; Caffeic Acids; Nitrates; Nitric Oxide; Platelet Aggregation Inhibitors; Rabbits

Novel nitric oxide (NO) releasing derivatives (7a-7l) of 3-n-butylphthalide (NBP) were designed and synthesized. Compound 7e inhibited the adenosine diphosphate (ADP), thrombin (TH) and arachidonic acid (AA)-induced in vitro platelet aggregation, superior to NBP and aspirin, released moderate levels of NO, and improved aqueous solubility relative to NBP. Furthermore, 7e exhibited greater antithrombotic activity than NBP and aspirin in rats, and protected against collagen and adrenaline-induced thrombosis in mice. Therefore, NO-releasing NBP derivatives possessed potent antiplatelet aggregation and antithrombotic activity. Our findings may aid in the design of new therapeutic agents for the treatment of thrombosis-related ischemic stroke.

Topics: Animals; Benzofurans; Blood Platelets; Drug Synergism; Fibrinolytic Agents; Male; Mice; Nitric Oxide; Platelet Aggregation Inhibitors; Rats; Rats, Sprague-Dawley; Thrombosis

The synthesis and sensory evaluation of enantiomeric sets of sedanenolide (1) and 3-butylphthalide (3) are described. The asymmetric synthesis was achieved via the intramolecular Diels-Alder reaction of chiral propargylester (5) which was prepared from optically active propargyl alcohol (4) and 2,4-pentadienoic acid. The sensory evaluation of these enantiomers revealed that there were distinct differences between their aroma character and odor threshold.

Topics: Alkynes; Apium; Benzofurans; Cyclization; Food Additives; Humans; Magnetic Resonance Spectroscopy; Molecular Structure; Odorants; Propanols; Sensory Thresholds; Stereoisomerism; Taste; Taste Perception

To study the chemical constituents of Ligusticum sinense.. The chemical constituents were isolated and purified by column chromatography on silica gel and Sephadex LH-20; Using spectroscopy methods to elucidate their structures.. Nine compounds were isolated and identified as levistolide A (1), (Z)-3-butylidene-7-hydroxyphthalide (2), senkyunolide B (3), 3-butylphthalide(4), (Z)-ligustilide (5), riligustilide (6), neocnidilide (7), senkyunolide A (8), beta-sitostesol (9).. Compounds 2 and 3 are obtained from this plant for the first time.

Topics: Aldehydes; Benzofurans; Ligusticum; Mass Spectrometry; Molecular Structure; Plant Extracts; Plants, Medicinal; Rhizome

The development of efficient methods for the facile construction of important molecular frameworks is an important goal in organic synthesis. Chiral 3-substituted phthalides are widely distributed in a large collection of natural products with broad, potent, and potentially path-pointing biological activities. In this investigation, we have uncovered an unprecedented organocatalytic asymmetric aldol-lactonization reaction of 2-formylbenzoic esters with ketones/aldehydes for convenient construction of the enantioenriched "privileged" scaffold. As a result of the sensitive nature of substrate structures of an organocatalytic enantioselective aldol reaction, after extensive optimization of reaction conditions, catalyst L-prolinamide alcohol IV is identified as the best promoter. Interestingly, it is found that in this reaction, addition of an acid additive PhCO(2)H can significantly enhance reaction efficiency with use of only as low as 2.5 mol % IV for the process. Moreover, due to the sensitivity of reaction conditions toward a sequential aldol-lactonization process without affecting enantioselectivity and racemization, it is essential to remove the catalyst for the subsequent facile lactonization reaction in the presence of K(2)CO(3). The aldol-lactonization processes serve as a powerful approach to the preparation of synthetically and biologically important 3-substituted phthalides with a high level of enantioselectivities. A 3-step catalytic asymmetric synthesis of the natural product of 3-butylphthalide is reported.

Topics: Aldehydes; Benzofurans; Catalysis; Cyclization; Ketones; Magnetic Resonance Spectroscopy; Molecular Structure; Stereoisomerism; Structure-Activity Relationship

A series of n-butylphthalide derivatives were designed and synthesized. The in vitro activities of these compounds were evaluated by a resting tension of isolated rat thoracic aorta ring assay. Compounds 4g and 4i were found to be more active than n-butylphthalide.

Topics: Animals; Aorta, Thoracic; Benzofurans; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Mice; Molecular Structure; Rats; Stereoisomerism; Vasodilator Agents

To determine the expression of glial fibrillary acidic protein(GFAP) and vascular endothelial growth factor(VEGF) in the hippocampus of rats with Alzheimer's disease(AD), and to determine the effect of butylphthalide on them and its significance.. Sixty male adult rats were randomly divided into a model group, a Butylphthalide group, and a control group. AD models were established by injecting beta-amyloid protein 1-42 into the hippocampus of rats. Sixty days later,the rats were sacrificed and both sides of the hippocampus were sectioned for immunohistochemistry.. Positive cells of GFAP in the hippocampus of the model group increased and the expression of VEGF decreased statistically, compared with the control group(P<0.01). The positive cells of GFAP in the hippocampus of the butylphthalide group decreased and the expression of VEGF increased significantly, compared with the model group(P<0.05).. Butylphthalide may protect the neuron-vascular unit of the hippocampus of Alzheimer model rats by inhibiting the expression of GFAP and increasing the expression of VEGF.

Topics: Alzheimer Disease; Animals; Benzofurans; Disease Models, Animal; Drugs, Chinese Herbal; Glial Fibrillary Acidic Protein; Hippocampus; Male; Random Allocation; Rats; Rats, Sprague-Dawley; Vascular Endothelial Growth Factor A

dl-NBP is a potentially beneficial and promising drug for treatment of ischemic stroke with multiple actions that affect different pathophysiologic processes, such as improving microcirculation, decreasing brain infarct volume, regulating energy metabolism, and especially inhibiting platelet aggregation and reducing thrombus formation. However, NBP is limited to use in the clinic by other side effects, such as elevated aminotransferase, abnormal liver function and digestive response. Some derivates of NBP were synthesized with the halides (F, Cl and Br) on the 6-position, and their IR and Raman spectra were measured. They proved the complemental information for deducing their structure. By comparing the spectra of the NBP, the band of disubstituted benzene disappeared in the derivatives, and the band of trisubstituted benzenes were observed. The stretching vibrational band of C--H was detected in the Raman spectra, but was not observed in IR. In the low frequency region, the deformation vibration band of --C--C--C--C was also observed in the Raman spectra.

Topics: Benzofurans; Spectrophotometry, Infrared; Spectrum Analysis, Raman

The aim of the present study was to explore the neuroprotective effects and mechanisms of action of dl-3-n-butylphthalide (NBP) in a 1-methyl-4-phenylpyridiniumion (MPP(+))-induced cellular model of Parkinson's disease (PD). NBP was extracted from seeds of Apium graveolens Linn. (Chinese celery). MPP(+) treatment of PC12 cells caused reduced viability, formation of reactive oxygen, and disruption of mitochondrial membrane potential. Our results indicated that NBP reduced the cytotoxicity of MPP(+) by suppressing the mitochondrial permeability transition, reducing oxidative stress, and increasing the cellular GSH content. NBP also reduced the accumulation of alpha-synuclein, the main component of Lewy bodies. Given that NBP is safe and currently used in clinical trials for stroke patients, NBP will likely be a promising chemical for the treatment of PD.

Topics: 1-Methyl-4-phenylpyridinium; alpha-Synuclein; Animals; Antioxidants; Apium; Benzofurans; Cell Survival; Cytoprotection; Glutathione; Membrane Potential, Mitochondrial; Mitochondria; Oxidative Stress; Parkinson Disease, Secondary; PC12 Cells; Rats; Reactive Oxygen Species

The aim of this study was to investigate the protective effect of dl-3-n-butylphthalide (NBP) on chronic brain injury caused by diabetes.. A group of diabetic Sprague-Dawley rats was orally treated with NBP for 6 weeks. In this study, we examined glial reactivity in hippocampus of streptozotocin (STZ)-induced diabetic rats by determining the expression of glial fibrillary acidic protein (GFAP) and CD11b. We also examined anti-apoptosis protein, vascular endothelial growth factor (VEGF) and key apoptosis enzyme, caspase-3, expression by immunohistochemistry.. We found that GFAP, CD11b, VEGF (685.1 +/- 35.5 cells/mm(2) in diabetic rats versus 320.6 +/- 21.9 cells/mm(2) in control rats, p<0.05, n=5) and VEGF(+)-caspase-3(+) (393.4 +/- 24.2 cells/mm(2) versus 135.8 +/- 12.0 cells/mm(2) in control rats, p<0.05, n=5) immunostaining increased in the hippocampus of diabetic rats; However, treatment with NBP resulted in an obvious reduction of GFAP and CD11b-immunoreactive gliocytes in hippocampus. VEGF expression was up-regulated (837.2 +/- 20.1 cells/mm(2), n=5), while the caspase-3 expression was reduced (240.0 +/- 15.1 cells/mm(2), n=5) in the NBP-treated diabetes mellitus-NBP rats.. These results suggest that diabetes causes increased glial reactivity, apoptosis and compensatory VEGF expression, and NBP may have a protective effect for diabetic brain damage through enhancing VEGF expression to inhibit caspase-3 mediated apoptosis.

Topics: Animals; Apoptosis; Benzofurans; Cerebral Arteries; Diabetes Mellitus; Diabetes Mellitus, Experimental; Disease Models, Animal; Gliosis; Hippocampus; Male; Neurons; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Stroke; Up-Regulation; Vascular Endothelial Growth Factor A

Alzheimer's disease (AD) is an age-related, progressive neurodegenerative disorder that occurs gradually and results in memory, behavior, and personality changes. L-3-n-butylphthalide (L-NBP), an extract from seeds of Apium graveolens Linn (Chinese celery), has been demonstrated to have neuroprotective effects on ischemic, vascular dementia, and amyloid-beta (Abeta)-infused animal models. In the current study, we examined the effects of L-NBP on learning and memory in a triple-transgenic AD mouse model (3xTg-AD) that develops both plaques and tangles with aging, as well as cognitive deficits. Ten-month-old 3xTg-AD mice were given 15 mg/kg L-NBP by oral gavage for 18 weeks. L-NBP treatment significantly improved learning deficits, as well as long-term spatial memory, compared with vehicle control treatment. L-NBP treatment significantly reduced total cerebral Abeta plaque deposition and lowered Abeta levels in brain homogenates but had no effect on fibrillar Abeta plaques, suggesting preferential removal of diffuse Abeta deposits. Furthermore, we found that L-NBP markedly enhanced soluble amyloid precursor protein secretion (alphaAPPs), alpha-secretase, and PKCalpha expression but had no effect on steady-state full-length APP. Thus, L-NBP may direct APP processing toward a non-amyloidogenic pathway and preclude Abeta formation in the 3xTg-AD mice. The effect of l-NBP on regulating APP processing was further confirmed in neuroblastoma SK-N-SH cells overexpressing wild-type human APP(695) (SK-N-SH APPwt). L-NBP treatment in 3xTg-AD mice also reduced glial activation and oxidative stress compared with control treatment. L-NBP shows promising preclinical potential as a multitarget drug for the prevention and/or treatment of Alzheimer's disease.

Topics: ADAM Proteins; Alzheimer Disease; Amyloid beta-Peptides; Amyloid beta-Protein Precursor; Animals; Benzofurans; Cell Line, Tumor; Cell Survival; Cognition Disorders; Disease Models, Animal; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Female; Humans; Indoles; Leukocyte Common Antigens; Male; Maleimides; Malondialdehyde; Maze Learning; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Nerve Tissue Proteins; Neuroblastoma; Neuroglia; Neuroprotective Agents; Oxidative Stress; Presenilin-1; Reaction Time; Spatial Behavior; Statistics, Nonparametric; tau Proteins; Time Factors; Transfection

Oxidative stress, an imbalance toward the prooxidant side of the prooxidant/antioxidant homeostasis, occurs in several brain neurodegenerative disorders. The protective effect of 3-butyl-6-bromo-1(3H)-isobenzofuranone (Br-NBP), a derivative compound of l-3-n-butylphthalide (NBP), on the hydrogen peroxide(H(2)O(2))- induced oxidative damage was investigated in PC12 cells. Following exposure of the cells to H(2)O(2), there was a significant reduction in cell survival,activities of glutathione peroxidase (GSH-px) and mitochondria membrane potential(MMP), in contrast, the increased levels in the leakage of lactate dehydrogenase (LDH), malondialdehyde (MDA) contents, nitric oxide (NO) productions, intracellular reactive oxygen species (ROS), intracellular Ca(2+) concentration ([Ca(2+)](i)), as well as cell apoptosis were observed. However, pretreatment of cells with Br-NBP prior to H(2)O(2) exposure attenuated all the changes mentioned above in a concentration-dependent manner except no effect on activities of GSH-px. Br-NBP exhibited the protective effect against H(2)O(2)-induced cytotoxicity in PC12 cells, suggesting that the compound may be a potential therapeutic agent for diseases induced by oxidative damage.

Topics: Animals; Apoptosis; Benzofurans; Calcium; Cell Survival; Drug Interactions; Glutathione Peroxidase; Hydrogen Peroxide; L-Lactate Dehydrogenase; Malondialdehyde; Membrane Potential, Mitochondrial; Neuroprotective Agents; Oxidants; Oxidative Stress; PC12 Cells; Rats; Reactive Oxygen Species

DL-3-n-Butylphthalide (NBP) has shown cytoprotective effects in animal models of stroke and has passed clinical trials as a therapeutic drug for stroke in China. Hence, as a potential clinical treatment for stroke, understanding the mechanism(s) of action of NBP is essential. This investigation aimed to delineate the cellular and molecular mechanism of NBP protection in neuronal cultures and in the ischemic brain. NBP (10 μM) attenuated serum deprivation-induced neuronal apoptosis and the production of reactive oxygen species (ROS) in cortical neuronal cultures. Adult male 129S2/sv mice were subjected to permanent occlusion of the middle cerebral artery (MCA). NBP (100 mg/kg, i.p.) administrated 2 hrs before or 1 hr after ischemia reduced ischemia-induced infarct formation, attenuated caspase-3 and caspase-9 activation in the ischemic brain. TUNEL-positive cells and mitochondrial release of cytochrome c and apoptosis-inducing factor (AIF) in the penumbra region were reduced by NBP. The proapoptotic signaling mediated by phospho-JNK and p38 expression was downregulated by NBP treatment in vitro and in vivo. It is suggested that NBP protects against ischemic damage via multiple mechanisms including mitochondria associated caspase-dependent and -independent apoptotic pathways. Previous and current studies and recent clinical trials encourage exploration of NBP as a neuroprotective drug for the treatment of ischemic stroke.

Topics: Animals; Apoptosis; Benzofurans; Blotting, Western; Brain Ischemia; Caspase 3; Caspase 9; Enzyme Activation; Fluorescent Antibody Technique; In Situ Nick-End Labeling; Male; MAP Kinase Kinase 4; Mice; Neurons; Neuroprotective Agents; Signal Transduction

To determine the effect of butylphthalide on the expressions of protein disulfide isomerase (PDI) and P53 in the brain tissue of rats with Alzheimer's disease (AD).. Sixty male adult rats were randomly divided into AD model group, butylphthalide group and control group (n = 20). AD models were established by injecting beta-amyloid protein 1-42 into the hippocampus of rats. Sixty days later, the learning and memory abilities of the rats were evaluated using Y-maze test, and the expressions of PDI and P53 in the brain tissue of the rats were measured by immunohistochemistry.. Compared with the control group, the rats in AD model group exhibited significantly reduced learning and memory abilities, lowered expressions of PDI in the hippocampus and increased expression of P53 in the cortex (P > 0.01). In comparison with the model group, the rats in the butylphthalide group showed significantly increased PDI-positive cells in the hippocampus and decreased expression of P53 in the cortex (P < 0.01).. Butylphthalide improves the learning and memory abilities of rats with experimental AD, the mechanism of which may involve inhibition of P53 expression and enhancement of PDI expression in the brain tissues.

Topics: Alzheimer Disease; Animals; Apoptosis; Benzofurans; Brain; Disease Models, Animal; Learning; Male; Memory; Protein Disulfide-Isomerases; Random Allocation; Rats; Rats, Sprague-Dawley; Tumor Suppressor Protein p53

Novozyme 435 could be a highly efficient catalyst in the asymmetric acylation of (R,S)-3-n-butylphthalide in tetrahydrofuran-hexane solvents. The effect of various reaction parameters such as agitation velocity, water content, mixed media, temperature, concentration of Novozyme 435, molar ratio of acetic anhydride to (R,S)-3-n-butylphthalide, reaction time, enantiomeric excess of substrate (ee(S)), enantiomeric excess of product (ee(P)), and enantioselective ratio (E) were studied. Tetrahydrofuran markedly improved (R,S)-3-n-butylphthalide conversion, enantiomeric excess of remaining 3-n-butylphthalide, and enantiomeric ratio. The optimum media were 50% (v/v) tetrahydrofuran and 50% (v/v) hexane. Other ideal reaction conditions were an agitation velocity of 150 rpm, 0.4% (v/v) water content, temperature of 30 °C, 8 mg/mL dosage of Novozyme 435, 8:1 (0.4 mmol: 0.05 mmol) molar ratio of acetic anhydride to (R,S)-3-n-butylphthalide, and a reaction time of 48 hr. Under the optimum conditions, 96.4% ee(S) and 49.3% conversion of (R,S)-3-n-butylphthalide were achieved. In addition, enantiomeric excess of the product was above 98.0%.

Topics: Acylation; Benzofurans; Candida; Catalysis; Enzymes, Immobilized; Fungal Proteins; Furans; Lipase; Water

To study the protective effects of butylphthalide (NBP) on the Aβ(25-35)-induced apoptosis in PC-12 cells.. The apoptosis of PC-12 cell was analyzed by MTT assay, transmission electron microscope and PI method at different concentrations of NBP (0.1, 1.0, 10, 100 µmol/L) with the addition of Aβ or not. The expressions of Bcl-2 and cytochrome C (Cyt-C) were detected by Western blot.. As demonstrated by the MTT assay, the values of cell viability were 76.5% ± 1.1%, 84.2% ± 1.3%, 89.5% ± 1.3% and 81.9% ± 1.9% at various concentration (0.1, 1.0, 10, 100 µmol/L) of NBP respectively. The model group was 71.7% ± 1.4%. It was revealed that the former could significantly prevent the cell viability under the induction of Aβ(25-35) (P < 0.05). A pretreatment with 10 µmol/L NBP could significantly inhibit the decrease of viability under the induction of Aβ(25-35) (P < 0.05). PI showed that the apoptosis rate of the 10 µmol/L NBP treatment group was significantly lower than that of the model group. Under electron microscope, the characteristics of cell apoptosis were significant in the model group. And the cell morphology of the 10 µmol/L NBP treatment group was normal. The expression rate of Bcl-2 protein in the 10 µmol/L NBP treatment group was obviously higher than that in the model group. Cyt-C was weakly expressed in nerve cells of the normal and the 10 µmol/L NBP groups. But it had a strong expression in the model group.. NBP prevents the injury of PC12 cells by Aβ. And the mechanism may be related to the elevated level of Bcl-2 and the inhibited mitochondrial release of Cyt-C.

Topics: Amyloid beta-Peptides; Animals; Apoptosis; Benzofurans; PC12 Cells; Peptide Fragments; Rats

To develop a RP-HPLC method for the simultaneous determination of five components including ligustrazine, ferulic acid, butylphthalide, ligustilide and butene-NBP in Rhizoma Chuanxiong, a Chinese herbal medicine.. The chromatographic analysis was carried out by using a grace smart RP C18 column with the mobile phase consisted of acetonitrile and 0.1% phosphoric acid (volume fraction), which was in gradient elution. A DAD detector was used to detect the components. The peak area was chosen under their maximum absorption wavelength for different components, and quantitated by using the external standard method.. The calibration curves were linear within the range of 0.008 35-0.668, 0.020 6-1.648, 0.012 2-0.976 , 0.050 75-4.06, 0.015 7-1.256 microg for ligustrazine, ferulic acid, butylphthalide, ligustilide and butene-NBP, respectively. The average recoveries of the five components were 101.98%, 99.91%, 96.94%, 100.85% and 99.04%, respectively.. This method is simple, quick, reproducible, with high recovery, and has been successfully applied to the simultaneous determination of the five components in Rhizoma Chuanxiong. This method can be used to control the quality of Rhizoma Chuanxiong and its preparations.

Topics: 4-Butyrolactone; Benzofurans; Calibration; Chromatography, High Pressure Liquid; Coumaric Acids; Drugs, Chinese Herbal; Pyrazines

It is shown that l-3-n-butylphthalide (l-NBP), the isomer of dl-NBP (racemic 3-n-butylphthalide, a new anti-cerebral ischemic agent) significantly attenuated cerebral hypoperfusion-induced learning dysfunction and brain damage in rats. In the present study, l-NBP (10 and 30 mg/kg) long-term (3-month) treatment of aged rat (21-month-old) significantly improved the learning and memory capability measured by the Morris water maze test. Hematoxylin-eosin-stained slices showed that both l-NBP at 30 mg/kg, and memantine as control at 20 mg/kg, attenuated the neurodegenerative changes in aged rats. L: -NBP treatment significantly increased the choline acetyltransferase activity and dose-dependently decreased the acetylcholinesterases activity in the hippocampus of aged rats. The immunohistological study demonstrated that expressions of beta-secretase and hyperphosphorylated tau protein were significantly increased in the hippocampus CA1 subfield and parietal cortex in aged rats. However, they were decreased significantly by treatment of l-NBP and memantine for 3 months. Our results indicated that long-term treatment with l-NBP might prevent age-related neurodegenerative changes by modulation of cholinergic system, reduction of phosphorylated tau and maintain structure and morphology of neurons. Therefore, l-NBP might be a potential drug for treatment of senile dementia.

Topics: Aging; Animals; Benzofurans; Dose-Response Relationship, Drug; Female; Male; Nerve Degeneration; Rats; Rats, Wistar; Time Factors

The asymmetric acylation of (R, S)-3-n-butylphthalide could be efficiently catalyzed by Novozyme 435. The effect of various reaction parameters such as water activity, temperature, molar ratio of acetic anhydride to (R, S)-3-n-butylphthalide, and reaction time on the asymmetric acylation were studied. The optimums of the reaction parameters were water activity 0.62, temperature 30 degrees C, molar ratio of acetic anhydride to (R, S)-3-n-butylphthalide 8:1, and reaction time 48 h, respectively. Under the optimum conditions, enantiopure 3-n-butylphthalide with an optical purity of 95.7% enantiomeric excess and 49.1% yield could be obtained. Furthermore, the enantiomeric excess of product was over 98%.

Topics: Acylation; Benzofurans; Catalysis; Chromatography, High Pressure Liquid; Enzymes, Immobilized; Fungal Proteins; Hexanes; Kinetics; Lipase; Stereoisomerism; Temperature

Appropriate restoration of blood flow via angiogenesis is critical for the recovery from ischemic stroke. Previously, we reported that treatment with dl-3n-butylphthalide (NBP) increases the number of local potent cerebral microvessels. However, the underlying mechanism remained unclear. The present study was conducted to test whether NBP enhances post-ischemic cerebral angiogenesis via vascular endothelial growth factor (VEGF) and hypoxia induced factor-1 alpha (HIF-1 alpha). Stroke-prone renovascular hypertensive rats (RHRSP) were used to create middle cerebral artery occlusion (MCAO) model. NBP was given 80 mg/kg per d for 10 consecutive days, starting 12, 24, 48 and 72 h respectively after MCAO. Neurological function was assessed daily and infarct volume as well as the expressions of CD31, VEGF, HIF-1 alpha and bFGF was detected 13 days after MCAO. The administration of NBP starting within 24 h after MCAO enhanced recovery of neurobehavioral function, reduced infarct volume, increased the quantity of CD31 positive vessels, and up-regulated expressions of VEGF and HIF-1 alpha. These findings suggest that treatment with NBP within 24 h post-ischemic stroke rescues brain tissue by enhancing angiogenesis associated with up-regulation of VEGF and HIF-1 alpha expressions.

Topics: Angiogenesis Inducing Agents; Animals; Benzofurans; Brain Ischemia; Cerebral Cortex; Disease Models, Animal; Drugs, Chinese Herbal; Hypertension; Hypoxia-Inducible Factor 1, alpha Subunit; Immunohistochemistry; Neovascularization, Physiologic; Rats; Up-Regulation; Vascular Endothelial Growth Factor A

DL-3-n-butylphthalide (NBP) has been used for stroke treatment in China for years. Recently, we found that NBP can reduce the incidence of stroke and have protective action on cerebral microvessels, suggesting a direct action of NBP on endothelial cells. However, it is difficult to evaluate the direct action of NBP on endothelial cells in vivo because of the interactions of endothelial cells with other types of neuronal cells. Therefore, we investigated whether NBP protects against oxygen glucose deprivation (OGD)-induced cell injury in an immortalized human umbilical vein endothelial cells (HUVEC) in vitro. Cells were exposed to OGD, leading to endothelial damage. Endothelial injury was assessed by measuring MTT and the changes in chromatin morphology. Mitochondrial superoxide, mitochondrial membrane potential and mitochondrial morphology were assessed using MitoSOX Red. Rhodamine 123 and MitoTracker, respectively. Nitrosative stress was assessed by measuring the production of peroxynitrite. The activity of superoxide dismutase (SOD) is evaluated using SOD assay kit-WST. The expression of hypoxia inducible factor-1 alpha (HIF-1alpha) was assessed at the protein level by immunofluorescence and Western blotting. NBP at doses between 0.01 and 100 micromol/L dose-dependently protected against OGD-induced cell death. In addition, NBP attenuated OGD-induced mitochondria superoxide, cellular formation of peroxynitrite, and decrease in SOD activity, mitochondria fragmentation and loss of mitochondrial membrane potential. In parallel, NBP enhanced OGD-induced HIF-1alpha expression. This study demonstrates that NBP can protect HUVEC against OGD-induced oxidative/nitrosative stress, mitochondrial damage and subsequent cell death. This protective effect is, at least in part, associated with its enhancement on OGD-induced HIF-1alpha expression.

Topics: Benzofurans; Blotting, Western; Cell Death; Cell Hypoxia; Cells, Cultured; Endothelial Cells; Fluorescent Antibody Technique; Glucose; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Mitochondria; Neuroprotective Agents; Oxidative Stress; Oxygen; Superoxide Dismutase

To investigate the effect of dl-3n-butylphthalide (NBP) on the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in protein and mRNA levels in the treatment of cerebral infarction with transient middle cerebral artery occlusion (MCAO) in rats.. The model of transient MCAO was established using the suture method of Longa by blocking middle cerebral artery (MCA) with a nylon suture. The MCA blood flow was restored by the withdrawal of the nylon suture 2 h after occlusion. Sham-operated rats (n=20) were prepared in similar fashion, but without occlusion of the MCA. Operated rats were randomizely divided into 2 groups (n=20 for each): vehicle group rats were only administered vegetable oil 2 mL twice daily for 3 days and NBP group rats were administrated NBP 25 mg/kg twice daily for 3 days. The infarct volume and neurological deficit were determined by TTC staining and Longa's score. VEGF and bFGF protein and mRNA were detected by immunohistochemistry and in situ hybridization.. NBP markedly inhibited the neurological deficit and reduced the infarct volumes as compare to vehicle group (P<0.05). NBP significantly upregulated VEGF and bFGF expression in both protein and mRNA levels in the peripheral infarct and hippocampus regions in contrast with sham-operated and vehicle group (P<0.05).. NBP has protective effects for cerebral ischemia through upregulating the expression of VEGF and bFGF.

Topics: Animals; Benzofurans; Fibroblast Growth Factor 2; Infarction, Middle Cerebral Artery; Ischemic Attack, Transient; Male; Neuroprotective Agents; Random Allocation; Rats; Rats, Sprague-Dawley; RNA, Messenger; Vascular Endothelial Growth Factor A

To determine the effect of butylphthalide on the expressions of p38 mitogen-activated protein kinase and extra-cellular signal regulated kinases (ERKs) in the brain tissue of rats with Alzheimer's disease (AD).. Sixty male adult rats were randomly divided to AD model group, butylphthalide group, and control group (n=20). AD models were established by injecting beta-amyloid protein 1-42 into the hippocampus of rats. Sixty days later, the learning and memory abilities of the rats were evaluated using Y-maze test, and the expressions of p38 and ERKs in the brain tissue of the rats were measured by immunohistochemistry. RESULTS Compared with the control group, the rats in AD model group exhibited significantly reduced learning and memory abilities, increased expressions of P38 in the hippocampus and lowered expression of ERK in the cortex (P<0.01). In comparison with the model group, the rats in the butylphthalide group showed significantly decreased P38-positive cells in the hippocampus and increased expression of ERK in the cortex (P<0.01).. Butylphthalide improves the learning and memory abilities of rats with experimental AD, the mechanism of which may involve inhibition of P38 expression and enhancement of ERK expression in the brain tissues.

Topics: Alzheimer Disease; Animals; Benzofurans; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation, Enzymologic; Hippocampus; Male; Memory; Neuroprotective Agents; p38 Mitogen-Activated Protein Kinases; Rats; Rats, Sprague-Dawley

To determine the expression of S100-beta protein and glial fibrillary acidic protein (GFAP) in hippocampal astrocytes of rats with Alzheimer disease (AD) model rats, and observe the effect of butylphthalide on their expression.. Sixty male adult rats were randomized equally into model group, butylphthalide group, and control group, and in the former two groups, AD models were established by injecting beta-amyloid protein 1-40 into the hippocampus. Sixty days later, the rats were sacrificed and the bilateral hippocampuses were taken for immunohistochemistry.. The number of cells positive for S100 and GFAP in the hippocampus in butylphthalide group were significantly higher than that in the control group (P/0.01), but lower than that in the model group (P/0.05).. The expression of S100 protein and glial fibrillary acidic protein increased significantly in the hippocampal astrocytes of rats with AD, and butylphthalide can inhibit the increase of their expression.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Benzofurans; Disease Models, Animal; Glial Fibrillary Acidic Protein; Hippocampus; Male; Nerve Growth Factors; Neuroprotective Agents; Peptide Fragments; Random Allocation; Rats; Rats, Sprague-Dawley; S100 Calcium Binding Protein beta Subunit; S100 Proteins

Alzheimer's disease is the most common form of dementia. Amyloid-beta protein is considered as a key factor of pathogenesis of Alzheimer's disease. l-3-n-butylphthalide (L-NBP), an anti-cerebral ischemia drug, has been shown to have therapeutic effects in vascular dementia animal models. In the present study, we investigated the potential of L-NBP to protect against cognitive impairment, oxidative damage and neuropathological changes induced by intracerebroventricular infusion of amyloid-beta peptide in rats. Daily treatments of 10 and 30 mg/kg L-NBP significantly improved spatial learning deficits and attenuated working memory deficits in Morris water maze task. L-NBP partially reversed the reduction of glutathione peroxidase activities and decreased malondialdehyde levels in the cortex and hippocampus. Furthermore, L-NBP markedly inhibited amyloid-beta-induced neuronal apoptosis, possibly by blocking caspase-3 activation. In addition, L-NBP reduced activation of glycogen synthase kinase-3beta and tau protein phosphorylation. Our results demonstrate that L-NBP protects against amyloid-beta-induced neurodegeneration and cognitive decline in a rat model, suggesting that it may have potential as a therapy for Alzheimer's disease.

Topics: Amyloid beta-Peptides; Animals; Apoptosis; Benzofurans; Cerebral Ventricles; Cognition Disorders; Infusion Pumps; Learning; Male; Memory; Neurons; Neuroprotective Agents; Oxidative Stress; Peptide Fragments; Phosphorylation; Rats; Rats, Wistar; tau Proteins

A rapid, sensitive and specific high performance liquid chromatography-electrospray ionization tandem quadrupole mass spectrometry (HPLC-MS/MS) method was developed and validated for the determination of 3-n-butylphthalide in rat plasma. Following protein precipitation with acetonitrile, 3-n-butylphthalide and glipizide (internal standard, I.S.) were separated using a gradient elution program on a C18 column and detected by mass spectrometry in positive ion mode with the multiple reaction monitoring (MRM) mode using the respective precursor to product ion combinations of m/z 191/145 for 3-n-butylphthalide and m/z 446/321 for glipizide, respectively. The total chromatographic running time was 2.5 min. The method was linear over the concentration range of 11.14-3480.00 ng/mL, using as little as 100 microL plasma. The lower limit of quantification (LLOQ) was 5.57 ng/mL. Finally, the method was successfully used to support a preclinical pharmacokinetic study of 3-n-butylphthalide in rats following intravenous administration.

Topics: Animals; Benzofurans; Chromatography, High Pressure Liquid; Male; Rats; Rats, Wistar; Sensitivity and Specificity; Tandem Mass Spectrometry

To explore the mechanism of action of butylphalide (NBP) against the injury following oxygen glucose deprivation/reoxygenation (OGD/R) in rat cortical neurons, neurons of Wistar newborn rats were prepared by filtering through a mesh, centrifugation and trypsogen digestion. A simple, stable and reliable in vitro model of OGD/R of neurons was established. We studied the activation, the nuclear translocation of NF-kappaB p65 and the mRNA expression of iNOS affected by NBP in each group neuron by RT-PCR. NBP is proved to be able to add cellular vigor and decrease LDH release. The mRNA expression of iNOS in neurons after OGD 4 h/R 8 h decreased when treated with NBP. There is statistical difference between each concentration of NBP that it adds cellular vigor, decreases LDH release and expression of iNOS in neurons after OGD 4 h/R 8 h. There is also statistical difference between NBP (100 micromol x L(-1)) and PDTC (100 micromol x L(-1)). It is proved that NBP can protect neurons, block upregulation of iNOS mRNA, and restrain activation of NF-kappaB in neurons.

Topics: Animals; Animals, Newborn; Antioxidants; Benzofurans; Cell Hypoxia; Cerebral Cortex; Drugs, Chinese Herbal; Female; Glucose; L-Lactate Dehydrogenase; Male; Neurons; Neuroprotective Agents; Nitric Oxide Synthase Type II; Pyrrolidines; Rats; Rats, Wistar; RNA, Messenger; Thiocarbamates; Transcription Factor RelA

Novozyme 435 could catalyze efficient acylation of 3-n-butylphthalide in organic medium. The conversion of 3-n-butylphthalide increased with the increase of hydrophobicity of solvent below that of hexane. The more available solvent was hexane. Salt hydride could control fixed water activity. The optimum water activity was 0.62. And the optimum of reaction time, velocity of agitation, dosage of Novozyme 435 and acetic anhydride to 3-n-butylphtrhalide molar ratio were 48 hours, 150 rpm, 8 mg/mL and 8:1, respectively. The conversion of 48.9% could be obtained at a water activity of 0.62 in hexane. Furthermore, Novozyme 435 had an enantioselective acylation of racemic 3-n-butylphthalide by original analysis.

Topics: Acylation; Benzofurans; Catalysis; Enzymes, Immobilized; Fungal Proteins; Kinetics; Lipase; Organic Chemicals

l-3-n-Butylphthalide (l-NBP), as an anti-cerebral ischemia agent, has been shown to have therapeutic effects on learning and memory deficits induced by chronic cerebral hypoperfusion and Abeta intracerebroventricular infusion in rats. In the present study, we investigated the neuroprotective effects of l-NBP on beta-amyloid (Abeta)25-35-induced neuronal death/apoptosis and potential mechanisms in rat hippocampal neurons and human neuroblastoma SH-SY5Y cells. Abeta25-35 significantly reduced cell viability and increased the number of apoptotic-like cells, indicating that Abeta25-35-induced neurotoxicity. In addition, tau protein hyperphosphorylation was found to increase after Abeta exposure. All of these phenotypes induced by Abeta25-35 were markedly reversed by l-NBP. Pretreatment with l-NBP prior to Abeta25-35 exposure significantly elevated cell viability, and reduced Abeta25-35-induced nuclear fragmentation and early apoptosis. Furthermore, immunoreactivity for hyperphosphorylation tau protein was significantly decreased by l-NBP treatment. Our results suggest that l-NBP may protect neurons against Abeta-induced neurotoxicity via inhibiting tau protein hyperphosphorylation.

Topics: Amyloid beta-Peptides; Animals; Apium; Apoptosis; Benzofurans; Brain Neoplasms; Cell Line, Tumor; Cell Survival; Drug Interactions; Hippocampus; Humans; Neuroblastoma; Neurons; Neuroprotective Agents; Peptide Fragments; Plant Extracts; Rats; Rats, Sprague-Dawley

To study the effects of dl-3n-butylphthalide (NBP) on the protein and mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats brain with permanent middle cerebral artery occlusion (MCAO).. The model of permanent MCAO was established by using the suture method of Longa, with which the nylon suture was used to make rat middle cerebral artery (MCA) blocked. Sham-operated rats (n=20) were prepared in similar fashion, but without doing the closed occlusion of the MCA. Operated rats were randomizely divided into model control and NBP groups (n= 20 for each). By intragastric administration, sham-operated and model control group rats were given vegetable oil 2 mL twice daily for 3 days, and also NBP group rats were given NBP 25 mg/kg twice daily for 3 days. The infarct volume and neurological deficit scores were determined by tetrazolium chloride (TTC) staining and Longa's score separately. The protein and mRNA of VEGF and bFGF were detected by immunohistochemistry and in situ hybridization.. NBP markedly inhibited the neurological deficit and reduced the infarct volumes as compared to model control group (P < 0.05). NBP significantly upregulated VEGF and bFGF expression in both protein and mRNA levels in the peripheral infarct and hippocampus regions in contrast with sham-operated and model control groups (P < 0.05). In the infarct core, the protein and mRNA levels of VEGF and bFGF did not show significantly any difference (P > 0.05).. NBP can significantly reduce neurological deficit and infarction volume, and therefore may have protective effect for cerebral ischemia through upregulating the expression of VEGF and bFGF.

Topics: Animals; Benzofurans; Brain; Brain Ischemia; Drugs, Chinese Herbal; Fibroblast Growth Factor 2; Infarction, Middle Cerebral Artery; Rats; Rats, Sprague-Dawley; Vascular Endothelial Growth Factor A

3-n-Butylphthalide (NBP) may be beneficial for the treatment of ischemic stroke with multiple actions on different pathophysiological processes. In the present study, we investigated the effect of NBP isomers on learning and memory impairment induced by chronic cerebral hypoperfusion in rats. Male Wistar rats were orally administered 10 and 30 mg/kg l-, d-, or dl-NBP daily for 23 days after bilateral permanent occlusion of the common carotid arteries. Rats receiving 10 mg/kg l-NBP performed significantly better in tests for spatial learning and memory, and they had attenuated cerebral pathology, including neuronal damage, white matter rarefaction, and glial activation compared with controls. Furthermore, 10 mg/kg l-NBP-treated rats had significantly higher choline acetyltransferase activity, decreased cortical lipid peroxide, and reduced hippocampal superoxide dismutase activity, compared with vehicle controls. However, d- and dl-NBP did not show significant beneficial effects. The present findings demonstrate that the beneficial effects of l-NBP on hypoperfusion-induced cognitive deficits may be due to preventing neuropathological alterations, inhibiting oxidative damage and increasing acetylcholine synthesis. Our results strongly suggest that l-NBP has therapeutic potential for the treatment of dementia caused by decreased cerebral blood flow.

Topics: Acetylcholinesterase; Animals; Astrocytes; Benzofurans; Brain Ischemia; Carotid Artery, Common; Catalase; Cerebral Cortex; Choline O-Acetyltransferase; Cognition Disorders; Corpus Callosum; Glutathione Peroxidase; Hippocampus; Learning Disabilities; Ligation; Male; Maze Learning; Memory Disorders; Neuroprotective Agents; Oxidative Stress; Rats; Rats, Wistar; Superoxide Dismutase; Visual Pathways

The purpose of the study is to establish a model of cold-induced stroke in hypertensive rats, and to study the preventive effect of dl-3n-butylphthalide ( NBP ) on stroke. Stroke-prone renovascular hypertension(RHRSP) was created in Sprague-Dawley rats. The animals were assigned randomly to NBP, aspirin treated and vehicle control group, with administration of the medications for 7 days, and then subjected to cold treatment in an environmentally controlled chamber for 3 days to induce the occurrence of stroke. The incidence of stroke, the volume of the brain lesion, patency of the microvessels by FITC-dextran perfusion and the number of microvessels by immunohisochemical detection of vwF were investigated. Cold induced different types of stroke in RHRSP. The incidence of ischemic stroke and the volume of the infarct were decreased, and the perfused microvessels were increased with NBP pretreatment. Our data suggest that NBP prevents cold-induced ischemic stroke via improvement of cerebral microvessels.

Topics: Animals; Aspirin; Benzofurans; Brain; Brain Ischemia; Cerebral Arteries; Cerebrovascular Circulation; Cold Temperature; Dextrans; Disease Models, Animal; Fluorescein-5-isothiocyanate; Hypertension, Renal; Male; Microcirculation; Molecular Structure; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; Stroke; Treatment Outcome

To study the expressions of BDNF, BDNF mRNA, NGF and NGF mRNA in the permanent focal cerebral ischemia tissues of rats. METHHODS: Healthy male Sprague-Dawley rats were taken for this study project. According to the procedure of Zea-Longa, the rat model with permanent cerebral ischemia was established by rat middle cerebral artery obstructed (MCAO) with a nylon thread, and the model rats of neurobehavioral evaluation as 1-3 grade were randomly divided into two groups: butylphthalide group (A group) and control group (B group). A group was given with 25 mg/kg butylphthalide, B group was given with edible oil, two times every day. 3 days after occlusion, all rats were sacrificed after evaluated the neurobehavioral scores, and the samples of cerebrum were obtained after in situ perfusion and fixation with 40 g/L paraformaldehyde. 5 rats in each group were taken to tetrazolium chloride (TTC) staining for macroscopic observation of cerebral infarction area, the rest samples were processed by immunohistochemistry to evaluate effects of butylphthalide on BDNF and NGF expression, hybridization in situ to evaluate effects of butylphthalide on BDNF mRNA and NGF mRNA expression. SPSS12. 0 for statistical analysis, it was P<0. 05 as having statistical significance.. Comparing to control group (B group), butylphthalide group (A group) did not have significantly pathological difference, but the grade of behavior and infarction area were apparently reduced (P<0. 05). In butylphthalide group, there was a significant expression up-regulation to BDNF, NGF, BDNF mRNA and NGF mRNA in the peripheral around infarction and cornu ammonis or hippocampus area (P<0. 05). However in the infarction area, the expressions of BDNF, NGF, BDNF mRNA and NGF mRNA had no significantly statistical difference (P> 0. 05).. Comparing to control group, butylphthalide can significantly up-regulate the expressions of BDNF and NGF in genetic transcription level, and protect from the ischemia injury.

Topics: Animals; Benzofurans; Brain-Derived Neurotrophic Factor; Cerebrum; Gene Expression Regulation; Ischemia; Male; Nerve Growth Factor; Neuroprotective Agents; Rats; Rats, Sprague-Dawley; RNA, Messenger; Stroke

A novel strategy for predicting bioactive components in traditional Chinese medicines (TCM) using live cell extraction and high performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS) analysis was proposed. The hypothesis is that when cells are incubated together with the extract of TCM, the potential bioactive components in the TCM should selectively combine with the cells, and the relative concentrations of the cell-combining components in the suspension medium should decrease, while the cell-combining components would be detectable in the extract of denatured cells. The identities of the cell-combining components could be determined by HPLC-DAD-MS analysis. Using the proposed approach, the potential bioactive components of Danggui Buxue decoction, a commonly used TCM for anaemia, and its compositions, Radix Angelica Sinensis and Radix Astragli for endothelial cells, were investigated. Six compounds in the extract of Danggui Buxue decoction were detected as the components selectively combined with endothelial cells, among them two were contributed by Radix Angelica Sinensis, and four by Radix Astragli. The identities of four of the six potential bioactive compounds were elucidated as ononoside, calycosin, 3-butylphthalide and ligustilide by HPLC-DAD-MS analysis. The results indicate that the proposed approach may be applied to predict the bioactive candidates in TCM.

Topics: 4-Butyrolactone; Benzofurans; Cell Extracts; Cells, Cultured; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Endothelial Cells; Humans; Isoflavones; Mass Spectrometry

2-(1-Hydroxypentyl)-benzoate (dl-PHPB), a derivate of 3-n-butylphthalide (dl-NBP), is a novel drug candidate used for treatment of cerebral ischemia. The goal of the present study was to investigate the effects of dl-PHPB on infarct volume, neurological function, and cerebral blood flow (CBF) in transient focal cerebral ischemia. Therefore, an animal model of 2-h middle cerebral artery occlusion (MCAO) followed by 24-h reperfusion was used. Rats received dl-PHPB (1.3, 3.9, or 12.9 mg/kg) intravenously 10 min after the onset of MCAO. Compared with the vehicle control group (37.4%), infarct volume in dl-PHPB-treated groups was reduced significantly and dose-dependently to 25.4, 17.4, and 13.7%, respectively. The changes in neurological deficient were also observed in neurobehavioral test in a dose-dependent manner, and the neuronal score was improved significantly from the vehicle control of 3.2 to 2.7, 2.1, and 1.8, respectively. At the highest dose, the potency of dl-PHPB was similar to those of dl-NBP. CBF was quantified by using laser-Doppler flowmetry. During the ischemia, the regional CBF values of dl-PHPB groups were significantly higher than that of vehicle group. In addition, our study showed that dl-PHPB converted into dl-NBP very quickly in blood in vitro. Approximately 70% of dl-PHPB converted into dl-NBP in 5 min when dl-PHPB was added into plasma at final concentrations of 6, 30, and 60 mug/ml. This result demonstrated that the neuronal protection effects of dl-PHPB were mainly induced by dl-NBP, an active compound converted from its precursor, dl-PHPB. In conclusion, dl-PHPB can reduce infarct volume and improve neurobehavioral deficits in a rat model of transient MCAO. Those effects may partially be due to an increase in CBF by the active metabolite (dl-NBP) of dl-PHPB. Therefore, our results suggest that dl-PHPB may be useful for treatment of ischemia stroke.

Topics: Animals; Benzoates; Benzofurans; Cerebrovascular Circulation; Disease Models, Animal; Dose-Response Relationship, Drug; Infarction, Middle Cerebral Artery; Ischemic Attack, Transient; Laser-Doppler Flowmetry; Male; Neuroprotective Agents; Pentanes; Prodrugs; Rats; Rats, Sprague-Dawley

3-n-butylphthalide (NBP) is a potentially beneficial drug for the treatment of ischemic stroke with multiple actions on different pathophysiological processes. In the present study, the effect of l-, d-, and dl-NBP was investigated on ADP-, collagen-, and AA-induced platelet aggregation. l-NBP was the most potent among l-, d-, and dl-NBP. At higher concentration the effect of dl-NBP on platelet aggregation was greater than that of l- or d-NBP alone. The ex vivo antiaggregatory activity of l-NBP 100mg/kg declined gradually after 2 hours, but a considerable antiplatelet activity was still observed 4h after l-NBP administration. NBP was given orally and resulted in a dose-dependent inhibition of thrombus formation. Of the two isomers, l-NBP was the most potent. It significantly protected mice from a mixture of collagen and epinephrine induced thromboembolic death. When 100 mg/kg of l-NBP were administered orally to rats, the bleeding time increased 2.1-fold compared with the control group. At the same dose, ex vivo platelet aggregation induced by ADP, collagen, and AA was inhibited by l-NBP and the antithrombotic effects of the compound were also observed. Thus, NBP exerts oral anti-platelet and anti-thrombotic efficacy without perturbing systemic hemostasis in rats. l-NBP is more potent than d- and dl-NBP as antiplatelet agent.

Topics: Animals; Benzofurans; Bleeding Time; Fibrinolytic Agents; Male; Mice; Platelet Aggregation; Platelet Aggregation Inhibitors; Rats; Rats, Wistar; Thrombosis

To investigate the effects of 3-n-butylphthalide (NBP) on apoptosis induced by transient focal cerebral ischemia in rats, compare the action potency of s-(-)-, r-(+)- and (+/-)-NBP, and clarify the enantiomer that played a main role.. DNA fragmentation was detected by the terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) assay and gel electrophoresis. The expression of cytochrome c and caspase-3 protein was observed by Western blot analysis and immunohistochemistry. Middle cerebral artery was occluded for 2 h.. Significant DNA fragmentation was detected at 24 h after reperfusion. This response was inhibited by s-(-)-NBP (5, 10 mg/kg i.p.). s-(-)-NBP 10 mg/kg almost completely inhibited DNA fragmentation, whereas r-(+)- NBP 10 mg/kg showed less effect. (+/-)-NBP (20 mg/kg) showed an inhibitory effect between that of s-(-)-NBP (10 mg/kg) and r-(+)-NBP (10 mg/kg). During the apoptotic process, cytochrome c was released into the cytosol and caspase-3 was activated. This effect was markedly inhibited by s-(-)-NBP, and the action potency of r-(+)- and (+/-)-NBP on the changes of cytochrome c and caspase-3 protein was similar to that on DNA fragmentation.. NBP, especially its s-(-)-enantiomer, could potently reduce the release of cytochrome c, decrease the activation of caspase-3, and inhibit DNA fragmentation after transient focal cerebral ischemia. Our findings on the beneficial effects of NBP on cerebral ischemia-induced apoptosis might have important implications for the study and treatment of ischemic cerebrovascular diseases.

Topics: Animals; Apoptosis; Benzofurans; Caspase 3; Caspases; Cytochromes c; DNA Fragmentation; Drugs, Chinese Herbal; Ischemic Attack, Transient; Male; Neuroprotective Agents; Rats; Rats, Wistar; Stereoisomerism

The aim of the present study was designed to explore the effect of (+/-) -3-n-butylphthalide (NBP) on ATPase, anti-oxidant enzymes activities and lipid peroxidation of mitochondria and cerebral cortex in rats subjected to 24 hours of reperfusion following 2 hours of cerebral ischemia (tMCAO).. Activities of SOD (Superoxide Dismutase), GSH-Px (glutathione Peroxidase,) and CAT (Catalase), and MDA level of mitochondria or cortex were measured by using biochemical methods in tMCAO rats.. (1) The activities of mitochondrial Na+K(+)-ATPase, Ca(2+)-ATPase and Mg2+ ATPase were found to decrease significantly in the vehicle group (ischemia + saline). Pre-treatment with NBP (5, 10, 20 mg/kg, i.p.) 10 min before tMCAO markedly enhanced the activities of Na+K(+)-ATPase and Ca(2+)-ATPase, compared with vehicle group. (2) The activities of SOD and mitochondrial GSH-Px were decreased and MDA level increased in vehicle groups as compared with that in sham group (non-ischemia + saline). NBP (20 mg/kg, i.p.) significantly enhanced total mitochondrial SOD activity, and also enhanced cerebral cortex total SOD activity (in 5, 10, 20 mg/kg groups). However, it had no obvious effect on CuZn-SOD activity. NBP (20 mg/kg i.p.) markedly increased mitochondrial (but not in cerebral cortex) GSH-Px activity; NBP 10, 20 mg/kg markedly decreased mitochondrial MDA level compared with that in vehicle group (P < 0.05). (3) The action of raceme NBP on the increase of the activities of ATPase and antioxidative enzymes seemed to be beneficial than that of (-) -NBP or (+) NBP.. The results suggest that NBP improves energy pump and subsides oxidative injury which may contribute to its anti-neuronal apoptotic effect.

Topics: Adenosine Triphosphatases; Animals; Benzofurans; Cerebral Cortex; Drugs, Chinese Herbal; Glutathione Peroxidase; Ischemic Attack, Transient; Lipid Peroxidation; Male; Mitochondria; Neuroprotective Agents; Rats; Rats, Wistar; Reperfusion Injury; Superoxide Dismutase

To study the effects of dl-, l- and d-3-n-butylphthalide (NBP) on platelet aggregation and thrombus formation.. Thrombus formation was assessed by silk thread-induced thrombosis in arteriovenous shunt in rats. Rat platelet aggregation induced by adenosine diphosphate (ADP), arachidonic acid (AA), collagen and thrombin was detected in vitro. The generation of thromboxane B2 (TXB2) and the concentration of cAMP in rabbit platelets in vitro were studied with radioimmunological assay.. dl-, l-NBP (5, 10, 20 mg.kg-1 i.p.) exhibited a dose-dependent inhibitory effect on thrombus formation in rats, while d-NBP was not active. dl, l, d-NBP (3-100 mumol.L-1) inhibited platelet-rich plasma aggregation in vitro induced by ADP, collagen and AA, and all of them showed no effect on thrombin-induced platelet aggregation. In addition, dl, l-NBP (10-100 mumol.L-1) were found to increase [cAMP]i in dose-related fashion. In the meantime, only high concentration of l-NBP was found to decrease platelet TXA2 level. In addition, l-NBP (1-100 mumol.L-1) showed significant effect on inhibiting 5-HT release from platelets. In contrast, dl- and d-NBP showed no effect.. The results suggest that NBP is a potent antiplatelet drug, the mechanism of its antithrombotic and antiplatelet activity is related to its regulation of cAMP level and 5-HT release.

Topics: Animals; Benzofurans; Blood Platelets; Cyclic AMP; Fibrinolytic Agents; Male; Neuroprotective Agents; Platelet Aggregation; Rabbits; Random Allocation; Rats; Rats, Sprague-Dawley; Serotonin; Stereoisomerism; Thrombosis

To evaluate the degree of neutrophil infiltration into ischemic tissue after transient focal cerebral ischemia, and to examine the effects of chiral 3-n-butylphthalide (NBP) on this inflammatory process.. After a 24-h reperfusion following transient cerebral ischemia, two different techniques, histologic analysis and modified myeloperoxidase (MPO)-quantification method, were utilized to identify the infiltration of neutrophils into cerebral tissue following ischemia. The expression of intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-alpha(TNF-alpha) in the ischemic zone were observed by immunohistochemistry, Western blot, and in situ hybridization techniques.. In cerebral cortex area perfused by middle cerebral artery (MCA), MPO activity was greatly increased after 24 h of reperfusion in the vehicle group, and it correlated well with the infiltration of neutrophils. Administration of dl-, d-, and l-NBP (20 mg.kg-1) partially inhibited both the increase in MPO activity and the appearance of neutrophils in ischemia-reperfusion sites. Up-regulation of ICAM-1 was also observed on the microvessel endothelium in the ischemic territory. In addition, chiral NBP markedly blunted ICAM-1 expression, and decreased the number of TNF-alpha blue purple-positive neurons induced by ischemia-reperfusion injury.. The results indicate that the increase in neutrophils infiltration into the infarct site implicated postischemic brain injury, and NBP was effective in protecting the ischemic sites following ischemic insult.

Topics: Animals; Benzofurans; Brain Ischemia; Cerebral Cortex; Infarction, Middle Cerebral Artery; Inflammation; Intercellular Adhesion Molecule-1; Leukocyte Count; Male; Neuroprotective Agents; Neutrophils; Peroxidase; Rats; Rats, Wistar; Reperfusion Injury; RNA, Messenger; Tumor Necrosis Factor-alpha

To explore if the inhibitory effect of 3-n-butylphthalide(NBP) on apoptosis induced by transient focal cerebral ischemia in rats is relevant to cortical calcineurin and calpain activities.. The model of cerebral ischemia-reperfusion was used. The activities of the two enzymes were measured by using biochemical methods.. DL-NBP and D-NBP 20 mg.kg-1 were found to significantly reduce ischemia ipsilateral cortical calcineurin and calpain activities. However, L-NBP 20 mg.kg-1 showed no obvious effect.. The anti-apoptotic effect of NBP may be relevant to its inhibition of calcineurin and calpain activities in focal cerebral ischemia rats.

Topics: Animals; Benzofurans; Brain Ischemia; Calcineurin; Calpain; Cerebral Cortex; Male; Neuroprotective Agents; Rats; Rats, Wistar; Reperfusion Injury

To investigate the protective effect of dl-3-n-butylphthalide (NBP) as an anti-cerebral ischemic drug on brain damage 24 h after closed head injury in mice.. Closed head injury was induced by dropping a 50-g weight from a height of 18 cm on a metal impounder resting on the parietal bone in mice.. The neurotraumatic model induced impairment of memory function, significant cerebral edema, and disruption of the blood-brain barrier. dl-3-n-butylphthalide (50 mg.kg-1) given intraperitoneally 5 minutes and 60 minutes after the onset of closed head injury was found to attenuate the impairment of memory function (P < 0.05), alleviate brain edema in the injured cerebral cortex (P < 0.05), and reduce extravasation of plasma protein bound to Evans blue dye by 63.5% (P < 0.01). NBP was also shown to increase the activity of choline acetyltransferase in the injured cortex to 0.83 +/- 0.21 ng.min-1.mg-1 (P < 0.01, compared with 0.48 +/- 0.14 ng.min-1.mg-1 of vehicle group).. NBP provides therapeutic response in experimental closed head injury.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Brain Edema; Brain Injuries; Head Injuries, Closed; Male; Mice; Neuroprotective Agents

To study the protective effect of dl-3-n-butylphthalide (NBP) on blood-brain barrier (BBB) damage induced by reperfusion following focal cerebral ischemia.. Focal cerebral ischemia in rats was performed by inserting a nylon suture into intracranial segment of internal carotid artery to block the origin of middle cerebral artery and reperfusion by withdrawing the nylon suture. Permeability of BBB was determined by extravasation of the protein-bound Evans blue dye to cerebral cortex and further evaluated by immunohistochemical or electronmicroscopic method.. Reperfusion for 3 h following focal cerebral ischemia for 3 h produced BBB damage which exhibited the increase in extravasation in cerebral cortex, elevation of the expression of immunoglobulin (IgG), and pore formation in endothelial cell membrane of capillary in cerebral cortex. NBP (5-20 mg.kg-1) decreased the extravasation in a dose-dependent manner. The expression of IgG in cerebral cortex was decreased and the ultrastructure damage of capillaries was alleviated after treatment with NBP. NBP 20 mg.kg-1 also alleviated brain edema caused by 3-h reperfusion following 3-h middle cerebral artery occlusion (MCAO).. NBP has protective effect on BBB damage induced by reperfusion after MCAO.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Capillaries; Capillary Permeability; Cerebral Cortex; Immunoglobulin G; Ischemic Attack, Transient; Male; Microscopy, Electron; Neuroprotective Agents; Rats; Rats, Wistar; Reperfusion Injury

To investigate the effect of dl-3-n-butylphthalide (dl-NBP) on experimental subarachnoid hemorrhage (SAH) in rats.. SAH was induced by injection of autologous artery blood 0.35 mL into lateral ventricle. Regional cerebral blood flow (rCBF) in caudate nucleus was determined by hydrogen clearance method.. A rapid and marked decrease in rCBF in caudate nucleus was observed 15 min after SAH and the rCBF remained at low level (about 50% pre-SAH value) within 180 min. dl-NBP (50, 100 mg.kg-1, i.g.) increased rCBF 30-180 min after the onset of SAH without significant effect on mean artery blood pressure. dl-NBP 100 mg.kg-1 increased rCBF in caudate nucleus by 26% at 15 min and by 36% at 180 min respectively after SAH. d-NBP but not l-NBP (10 mg.kg-1, i.p.) increased rCBF.. dl-NBP improves rCBF in caudate nucleus of rats subjected to SAH.

Topics: Animals; Benzofurans; Caudate Nucleus; Male; Neuroprotective Agents; Rats; Rats, Wistar; Regional Blood Flow; Subarachnoid Hemorrhage

The effects of dl-3-n-butylphthalide (dl-NBP), l-3-n-butylphthalide (l-NBP), and d-3-n-butylphthalide (d-NBP) on the production of nitric oxide (NO), epoprostenol (Epo) and endothelin-1 (ET-1) were investigated in cerebrovascular and aortic endothelium in culture.. Bovine cerebral endothelial cells (BCEC) and bovine aortic endothelial cells (BAEC) were cultured in Medium 199 in vitro. After incubation with dl-, l-, and d-NBP for 24 h, the release of NO, Epo, and ET-1 were analyzed by using spectrometry assay and radioimmunoassay (RIA) respectively.. Low concentrations of dl- and l-NBP (0.1-10 mumol.L-1) enhanced nitrite and 6-ketoprostaglandin F1 alpha (6-ketoPGF1 alpha) production in both BAEC and BCEC after a 24-h incubation, and l-NBP has a potent effect on promoting Epo production in BCEC. The production of ET-1 secreted by BCEC and BAEC was increased after TNF alpha stimulation, this enhancement was not blunted by the simultaneous addition of dl-, l-, and d-NBP.. 1) dl-NBP and l-NBP increase NO production in both BCEC and BAEC. 2) l-NBP increases more Epo production in BCEC than that in BAEC, and dl-NBP has selective effect on increasing Epo production in BCEC.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Animals, Newborn; Aorta; Benzofurans; Cattle; Cells, Cultured; Cerebral Cortex; Endothelin-1; Endothelium, Vascular; Epoprostenol; Neuroprotective Agents; Nitric Oxide; Stereoisomerism

To study the effect of dl-3-n-butylphthalide (NBP) on regional cerebral blood flow (rCBF) in forcal cerebral ischemia rats.. In chloral hydrate-anesthetized rat, the proximal portion of right middle cerebral artery (RMCA) was occluded, and H2 needle electrode was implanted in right striatum. rCBF was monitored in striatum using hydrogen clearance method.. Ten min after RMCA occlusion (RMCAO), NBP (5, 10, 20 mg.kg-1 i.p.) markedly increased rCBF to striatum (P < 0.01). When NBP was given i.p. 40 min after RMCAO, the increasing effect on rCBF was also observed (P < 0.05). However, when NBP was injected i.p. 60 min after RMCAO, the increasing effect of NBP on rCBF was not found. In NBP-pretreated (i.p. 40 min before RMCAO) group, rCBF in striatum measured at different time points of 30, 60, 90, 120, 150, and 180 min after RMCAO were increased by 97%, 107%, 136%, 211%, 173%, and 317%, respectively, compared with the percentages of vehicle group. The potency of the effect of Nim (0.5 mg.kg-1 i.p.) was similar to that of NBP (10 mg.kg-1 i.p.).. NBP pre-treatment or post-treatment markedly enhanced the rCBF to striatum in RMCAO rats.

Topics: Animals; Benzofurans; Cerebrovascular Circulation; Corpus Striatum; Drugs, Chinese Herbal; Ischemic Attack, Transient; Male; Neuroprotective Agents; Nimodipine; Rats; Rats, Wistar; Vasodilator Agents

To investigate the therapeutic effects of dl-3-n-butylphthalide (dl-NBP) on regional cerebral blood flow impairment and blood-brain barrier damage induced by subarachnoid hemorrhage (SAH) in rats.. SAH was induced by an injection of 0.35 ml autologous blood into the lateral ventricle. The regional cerebral blood flow in caudate nucleus was determined by hydrogen clearance method. Permeability of blood-brain barrier was measured by extravasation of the protein bound Evans blue dye to brain tissue.. SAH produced a marked decrease in both caudate blood flow and blood-brain barrier damage which showed increased extravasation of protein bound Evans blue dye to brain tissue. dl-NBP (5-20 mg/kg, given 5 min after the onset of SAH) and nimodipine (0.25 mg/kg) significantly increased the regional cerebral blood flow in caudate nucleus within 3 hours of SAH. Moreover, dl-NBP (10 mg/kg) and nimodipine (0.25 mg/kg, given 5 min and 3 h after SAH) significantly reduced the brain extravasation of Evans blue dye which increased after 6 hours of SAH.. By improving the regional cerebral blood flow and blood-brain barrier, dl-NBP has therapeutic effects on experimental SAH.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Caudate Nucleus; Male; Neuroprotective Agents; Rats; Rats, Wistar; Regional Blood Flow; Subarachnoid Hemorrhage

Effects of dl-3-n-butylphthalide(dl-NBP) on changes of regional cerebal blood flow (rCBF) and blood-brain barrier of rats subjected to subarachnoid hemorrhage (SAH) were investigated. Regional CBF was determined by hydrogen clearance method at 15 min before, 15 and 30 min thereafter once every 30 min up to 180 min after the beginning of SAH. The results showed that SAH produced a significant decrease in rCBF in caudate nucleus and blood-brain barrier damage which displayed increased extravasation of Evans blue to brain tissue. Both dl-NBP (5-20 mg.kg-1 given intraperitoneally 5 min after the onset of SAH) and nimodipine (0.25 mg.kg-1) were shown to increase rCBF in caudate nucleus through 3 h of SAH. Moreover, dl-NBP 10 mg.kg-1 and nimodipine 0.25 mg.kg-1, given 5 min and 3 h, respectively, after SAH, significantly reduced the brain extravasation of Evans blue 6 h after SAH, indicating that both dl-NBP and nimodipine has protective effects on blood-brain barrier. These results suggest that dl-NBP has therapeutic effects on SAH.

Topics: Animals; Benzofurans; Blood-Brain Barrier; Cerebrovascular Circulation; Drugs, Chinese Herbal; Male; Neuroprotective Agents; Rats; Rats, Wistar; Subarachnoid Hemorrhage

The effects of 3-n-butylphthalide(NBP) on the levels of 6-keto-PGF1 alpha, TXB2 and 6-keto-PGF1 alpha/TXB2 ratio were studied with methods of RIA. d-NBP and l-NBP(0.1-100 mumol.L-1) concentration-dependently increased 6-keto-PGF1 alpha release, decreased TXB2 release from neuronal cells, and significantly enhanced extracellular 6-keto-PGF1 alpha/TXB2 ratio in primary cultured rat cortical neurons exposed to hypoxic-hypoglycemic media for 5 h or hypoxic-hypoglycemic media for 5 h following normal media for 3 h. Aspirin(0.1-100 mumol.L-1) was also shown to inhibit TXB2 release from cortical neurons in a dose-dependent manner. However aspirin only increased 6-keto-PGF1 alpha/TXB2 ratio at low dose because aspirin inhibited both 6-keto-PGF1 alpha and TXB2 release simultaneously at large dose(10-100 mumol.L-1). This suggests that the action of l-NBP, d-NBP and dl-NBP on the increase of 6-keto-PGF1 alpha/TXB2 ratio might be one of the mechanisms in which NBP enhanced focal cerebral blood flow and improved ischemic brain damage.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Benzofurans; Cell Hypoxia; Cells, Cultured; Cerebral Cortex; Drugs, Chinese Herbal; Female; Fetus; Glucose; Neurons; Neuroprotective Agents; Rats; Rats, Wistar; Thromboxane B2

The effects of l-3-n-butylphthalide(l-NBP) and d-3-n-butylphthalide(d-NBP) on hypoxia/hypoglycemia-induced cytotoxicity in primary cultured rat cortical neurons were studied. l-NBP and d-NBP(1-100 mumol.L-1) were shown to inhibit hypoxia/hypoglycemia-induced LDH release, decrease the percent of cell death and improve the damaged cellular morphology at 10 mumol.L-1 concentration. In addition, l-NBP, d-NBP and dl-NBP were also found to significantly reduce the liberation of polyribosomes from the neuronal rough endoplasmic reticulum and disaggregation of polyribosomes induced by hypoxia/hypoglycemia. These data suggest that l-NBP, d-NBP and dl-NBP can remarkably protect cultured neurons against hypoxia/hypoglycemia induced damage.

Topics: Animals; Benzofurans; Cell Death; Cell Hypoxia; Cells, Cultured; Cerebral Cortex; Female; Fetus; Hypoglycemia; L-Lactate Dehydrogenase; Neurons; Neuroprotective Agents; Polyribosomes; Rats; Rats, Wistar; Stereoisomerism

Transient cerebral ischemia may cause striking changes in gene expression in rat brain. The induction of heat shock protein 70 (hsp70) mRNA is considered to be an important marker of cerebral ischemia injury, and c-fos may upregulate the expression of other genes related to the secondary injuries. dl-3-n-Butylphthaline(NBP) had been shown to have good anti-cerebral ischemic effect. Using the in situ hybridization and Northern blot technique, the effect of NBP on the expression of hsp70 mRNA and c-fos in transient middle cerebral artery occlusion (MCAo) rat caused by intraluminal thread was studied, and found that the expression of hsp70 mRNA was at the lesioned site at 1 h of reperfusion. It increased gradually with the duration of reperfusion time and peaked at 12 h at the lesioned site. With NBP treatment(i.p. 10 mg.kg-1 10 min before ischemia or 20 mg.kg-1 after ischemia), the expression of hsp70 mRNA attenuated significantly. For c-fos, the expression appeared at 0.5 h of reperfusion, peaked at 3 h, and decreased at 6 h. NBP pretreatment (10 mg.kg-1 10 min before ischemia) also decreased the c-fos expression. The same results were obtained with Northern blot technique. Since NBP had been shown to have good anti-cerebral ischemic effects, the attenuating effect on gene expression seemed to be the secondary effect after the alleviation of tissue injury.

Topics: Animals; Benzofurans; HSP70 Heat-Shock Proteins; Ischemic Attack, Transient; Male; Neuroprotective Agents; Proto-Oncogene Proteins c-fos; Rats; Rats, Wistar; RNA, Messenger; Stereoisomerism

The effects of l-3-n-butylphthalide(l-NBP) and d-3-n-butylphthalide(d-NBP) on extracellular nitric oxide (NO) levels and intracellular cyclic GMP (cGMP) levels were studied in primary cultured rat cortical neuronal cells. Nitric oxide and cGMP levels were measured by using spectrometry and radioimmunological analysis(RIA), respectively. The results showed that d-NBP (0.1-100 mumol: L-1) markedly increased extracellular NO levels and intracellular cGMP levels in primary cultured neurons that were exposed for 10 h to hypoxic/hypoglycemic, N-methyl-D-aspartate (NMDA), or KCl media. On the contrary, l-NBP(0.1-100 mumol.L-1) significantly decreased extracellular NO levels and intracellular cGMP levels. It is suggested that there is a contrary effect of d-NBP and l-NBP on NO release and cGMP production induced by hypoxia/hypoglycemia, NMDA, or KCl.

Topics: Animals; Benzofurans; Cells, Cultured; Cerebral Cortex; Cyclic GMP; Female; Fetus; Neurons; Neuroprotective Agents; Nitric Oxide; Rats; Rats, Wistar; Stereoisomerism

To study the effects of dl-3-n-butylphthalide (NBP) on the changes of thromboxane B2 (TXB2) and 6-keto-PGF1 alpha (6-keto-PGF1 alpha) contents in hippocampus, striatum, and cerebral cortex of rats subjected to focal cerebral ischemia followed by reperfusion.. Focal cerebral ischemia was induced by inserting a nylon suture into intracranial segment of internal carotid artery from external carotid artery and blockade of the origin of middle cerebral artery. For reperfusion, the suture was pulled out to restore the blood flow to the ischemic brain. Determination of TXB2 and 6-keto-PGF1 alpha was performed by RIA method.. Reperfusion following focal cerebral ischemia resulted in increases in TXB2 at 5 min and 6-keto-PGF1 alpha at 30 min and a decrease in the ratio of epoprostenol (PGI2)/thromboxane A2 (TXA2) (6-keto-PGF1 alpha/TXB2) at 5 min in hippocampus, striatum, and cerebral cortex. NBP 10 mg.kg-1 reduced the content of TXB2 without decreasing effect on 6-keto-PGF1 alpha. NBP 20 mg.kg-1 reduced both TXB2 and 6-keto-PGF1 alpha in lesser extent than aspirin (Asp, 20 mg.kg-1). NBP 20 or 10 mg.kg-1 elevated the ratio of PGI2/TXA2 after reperfusion, but Asp 20 mg.kg-1 did not increase the ratio except in striatum at 5 min after reperfusion.. NBP increases the ratio of PGI2/TXA2 which may have beneficial effects on the impaired microcirculation in postischemic brain tissues.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Aspirin; Benzofurans; Brain; Ischemic Attack, Transient; Neuroprotective Agents; Rats; Reperfusion Injury; Thromboxane B2

The present study evaluated the effect of dl-3-n-butylphthalide(NBP), a novel brain protective agent, on brain edema in rats following focal ischemia. Edema was induced by occluding the right middle cerebral artery (MCAO), producing permanent focal ischemia in the right cerebral hemisphere, which developed ipsilateral brain edema reproducibly. Edema was assessed 24 h after MCA occlusion by determining the brain water content from wet and dry weight measurements, and the sodium, potassium concentrations with ion-selective electrodes. In this model, NBP at the dose of 80, 160 and 240 mg/kg p.o. 15 min after MCAO prevented from brain edema in a dose-dependent manner. A significant reduction of sodium content and an increase in potassium level were observed in all drug-treated groups. It showed that NBP strongly attenuated brain water entry, sodium accumulation and potassium loss. Nimodipine treatment (5 mg/kg s.c.) also reduced brain edema (P < 0.05). The results suggest that a strong anti-edema activity of NBP may play an important role to contribute to the treatment of ischemic damage.

Topics: Animals; Benzofurans; Brain Edema; Drugs, Chinese Herbal; Hypoxia-Ischemia, Brain; Male; Neuroprotective Agents; Rats; Rats, Wistar

The protective effects of l-3-n-butylphthalide (l-NBP) and d-3-n-butylphthalide (d-NBP) on KCl (20 mmol.L-1)- or NMDA (N-methyl-D-aspartate, 30 mumol.L-1)-induced cytotoxicity in primary cultured rat cortical neurons were studied. Intracellular LDH release, percentage of cell death and cellular morphological changes were used to evaluate the effect of drugs. l-NBP (1-100 mumol.L-1) and d-NBP (1-100 mumol.L-1), but not nimodipine (1-100 mumol.L-1), were shown to dose-dependently inhibit LDH release induced by NMDA (30 mumol.L-1) in cultured rat cortical neurons with IC50 values of 4.89 mumol.L-1 and 13.52 mumol.L-1, respectively. The percent cell death was reduced with IC50 values of 44.37 and 49.78 mumol.L-1, and the cellular morphology improved. The effect seemed to be the same as that of equal concentration of NAME(NG-nitro-L-arginine methyl ester). In addition, l-NBP(10 mumol.L-1), d-NBP (10 mumol.L-1) and nimodipine(10 mumol.L-1) also produced significant inhibition on intracellular LDH release and decrease in percent cell death induced by KCl(20 mmol.L-1) in cultured neurons. The potencies of l-NBP and d-NBP were similar to that of equal dose of nimodipine. These data suggest that l-NBP and d-NBP can remarkably protect cultured neurons against the damage induced by KCl and NMDA.

Topics: Animals; Benzofurans; Cell Death; Cells, Cultured; Cerebral Cortex; L-Lactate Dehydrogenase; N-Methylaspartate; Neurons; Neuroprotective Agents; Potassium Chloride; Rats; Rats, Wistar; Stereoisomerism

The metabolites of dl-3-n-butylphthalide(NBP), a novel drug with promising protective action against cerebral ischemia, was studied in rats. Two main in vitro metabolites of NBP, M I and M II, were isolated and purified from rat liver microsome incubating system by using HPLC. The structure elucidation was mainly accomplished by spectral studies(UV, 1H-NMR, MS). Within 24 h following i.g. 3H-NBP, the total radioactivity excreted in urine and feces was 73.7% of the dose. Comparing with previous study, within 72 h following i.g. NBP, the total prototype drug excreted in urine and feces was 2.53% of the dose. This result excludes the possibility that NBP accumulates in vivo. The urine and brain homogenate of the rats(i.g. 3H-NBP) were analyzed by TLC. M I and M II were found in urine and M I was found in brain only. Furthermore, the ratio of radioactive M I to proptype drug was 1:1 in rat brain within 1 h following i.g. 3H-NBP. So, M I and M II were supposed to be the two main in vivo metabolites of NBP and M I might be an active metabolite.

Topics: Animals; Benzofurans; Brain; Feces; Male; Neuroprotective Agents; Rats; Rats, Wistar

The effects of NBP on concentrations of some purine metabolites in extracellular fluid of rat striatum during global ischemia and reperfusion were studied. Global ischemia was produced by the four-vessel occlusion method. Push-pull cannula was implanted stereotaxically into the striatum of rat and was perfused with Ringer's solution at a flow rate of 2.5 microliters.min-1. The level of adenosine(Ade), inosine(Ino), hypoxanthine(Hyp) and xanthine(Xan) in perfusates were measured with HPLC connected with a UV detector. The results indicate that the levels of ade, ino, hyp and xan were significantly increased (about 3-5 times of initial value) during cerebral ischemia and reperfusion. NBP at the dose of 20 or 40 mg.kg-1 given intra-peritoneally 20 min before ischemia was shown to depress the increase of ade, ino, hyp and xan during ischemia and reperfusion dose dependently. But no change in the level of purine metabolites was found in sham operated rats. It has been known that harmful free radicals were produced when xan and uric acid were formed by xanthine oxidase during reperfusion. This might be important for the development of ischemic injuries. Our findings suggest that the effect of NBP might be beneficial for protection against post-ischemic neuronal damage.

Topics: Adenosine; Animals; Benzofurans; Brain Ischemia; Corpus Striatum; Extracellular Space; Inosine; Male; Neuroprotective Agents; Rats; Rats, Wistar; Reperfusion; Stereoisomerism; Xanthines

Effects of NBP on liability of stroke, life span and neurological deficits following stroke were studied in stroke prone spontaneously hypertensive rats (SHRsp). The SHRsp rat was kept on 1% NaCl solution as drinking water and was fed 15 g soft food containing 0.6-0.8 g NaCl per day. Total NaCl intake for one rat was 1.1-1.3 g per day. After the onset of stroke, tap water and normal food was given instead of that containing NaCl. The neurological deficits were evaluated by a specially designed scoring system. These symptoms were divided into 4 degrees (1-4). Grade 1. stress (mild). Grade 2. forelimb or head twitch or with stress (severe). Grade 3. hemiparalysis, body inclined or disabled. Grade 4. paralysis, tremor or convulsion. Blood pressure, heart rate and body weight were measured once every 2 weeks. The weights of heart, brain and kidneys were also measured. The results show that NBP pre-treatment at the dose of 100 mg.kg-1.d-1 po delayed the onset of stroke. So, like nimodipine, NBP showed a stroke preventive action in SHRsp rats. In addition, treatment with NBP 100 mg.kg-1.d-1 po after the onset of stroke, the life span was prolonged and the score of neurological deficit decreased significantly. Because high blood pressure can not be lowered by NBP treatment, therefore, the protective effect against stroke can not be explained by the effect of hypotension. No change was found in BP, HR and the organ weight. The results indicate that NBP is expected to be useful in the treatment of stroke.

Topics: Animals; Benzofurans; Blood Pressure; Body Weight; Cerebrovascular Disorders; Heart Rate; Male; Neuroprotective Agents; Nimodipine; Rats; Rats, Inbred SHR; Stereoisomerism

1. Aqueous celery extract was administered intraperitoneally to genetically hypercholesterolaemic (RICO) and normocholesterolaemic (RAIF) rats via Alzet osmotic pumps over a 13 day period. 2. The serum cholesterol concentration of the celery extract-treated RICO rats was found to be significantly lower (P < 0.05) than the control rats. Aqueous celery extract was effective in preventing the rise of cholesterol level in the RICO rats. However, no such observation was seen in the RAIF rats. The serum triglyceride level was unchanged in both strains of rats. 3. When 3-n-butylphthalide (BuPh), a unique component in celery, was administered in the same manner to the RICO and RAIF rats, it did not produce significant changes in the serum and liver lipid profiles of these rats. The activities of hepatic 3-hydroxy-3-methylglutaryl CoA reductase in both strains of rats were also not significantly different from their respective controls. 4. Together with our recently reported thin-layer chromatography findings that BuPh was not detected in the aqueous celery extract, this study suggests that the effect of celery extract on serum cholesterol levels in the RICO rats could be attributed to chemical constituents other than BuPh.

Topics: Animals; Benzofurans; Cholesterol; Cholesterol, HDL; Hydroxymethylglutaryl CoA Reductases; Hypercholesterolemia; Lipoproteins; Liver; Plant Extracts; Rats; Triglycerides; Vegetables

The effect of dl-3-butylphthalide (NBP) on the contents of amino acids and dopamine in the rat striatum during globe cerebral ischemia has been studied. By using the technique of microperfusion in the striatum of rats subjected to 4-vessel occlusion cerebral ischemia, the extracellular contents of glutamate, taurine, gamma-aminobutyric acid and dopamine were found to be significantly increased in the striatum during the 20 min of cerebral ischemia. NBP (40 mg.kg-1; i.p. 30 min before ischemia) was shown to reduce the contents of dopamine and its metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), in the striatal extracellular fluid of the rat during ischemia. The content of glycine before and after ischemia was also reduced. However, no significant effect on the contents of glutamate and some other amino acids was observed. The results suggest that NBP may improve the striatal ischemic injury.

Topics: 3,4-Dihydroxyphenylacetic Acid; Animals; Benzofurans; Brain Ischemia; Corpus Striatum; Dopamine; Extracellular Space; Glycine; Male; Neuroprotective Agents; Rats; Rats, Wistar

A GC-MS method for the investigation on the metabolism of n-butyl phthalide (NBP) is described. After oral administration of NBP to rats, urine was collected, hydrolyzed with beta-glucuronidase, extracted and concentrated for TMS derivatization, and then analyzed by GC-MS. HBP and its four oxidative metabolites were determined in 0-24 h, 24-48 h rat urine. The mass spectra of the metabolites and their derivatives were presented and the in vivo metabolic pathway was discussed.

Topics: Animals; Benzofurans; Gas Chromatography-Mass Spectrometry; Male; Rats; Rats, Wistar

The effects of NBP on gasping and brain energy metabolism after complete brain ischemia in mice subjected to decapitation were investigated. The levels of ATP, phosphocreatine (PCr) and lactate were determined by the method of Lowry. The data indicated that NBP at 112.5 or 250 mg.kg-1 sc can significantly prolong the duration of gasping and at the dose of 150 or 200 mg.kg-1 sc reduce the level of lactate and increase the levels of ATP and PCr after complete brain ischemia. The results suggest that NBP may have brain protective action and improve ischemic brain energy metabolism.

Topics: Adenosine Triphosphate; Animals; Anticonvulsants; Benzofurans; Brain; Brain Ischemia; Decerebrate State; Energy Metabolism; Female; Lactates; Male; Mice; Phosphocreatine

dl-3-n-Butylphthalide (NBP) was known to have improving effect on brain energy metabolism after ischemia insult. The purpose of this study is to determine if the drug has protective action against ischemic neuronal damage. In the present study, the effect of NBP on cerebral infarction and neurological deficits after middle cerebral artery occlusion (MCAO) in rats was investigated. Focal cerebral ischemia was produced by permanent occlusion of the proximal portion of the right middle cerebral artery (MCA) according to the technique of Tamura. The infarct area was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining technique. The extent of neurological deficits was evaluated by the method of Bederson. The histological changes in neuronal change after MCAO in rats were also studied. The results indicate that the infarct area and the score of neurological deficits after MCAO were reduced significantly following intraperitoneal pretreatment or pre- and post-treatment with NBP 20 mg . kg-1. The treatment with NBP 10 or 20 mg . kg-1(i.p.), or 20,40 or 80 mg . kg-1 (po) 15 min and even 2 h (20 mg . kg-1, i.p.) after MCAO also markedly reduced the infarct area and the score of neurological deficits. However, no effect was found when NBP (20 mg . kg-1) was injected intraperitoneally 4 h after MCAO. MK-801 (0.5 mg . kg-1, i.p.), a non-competitive antagonist of NMDA receptor, significantly reduced the size of infarction and the score of neurological deficits in rats subjected to MCAO. The potency of NBP in reducing the infarct area and neurological deficits was found to be quite similar to that of MK-801 (0.5 mg . kg-1, i.p.). No neuroprotective effect of nimodipine (1.0 mg . kg-1, sc) was found. Generally, the potency of NBP in protecting rats from ischemic neurological damage is equal to that of MK-801 and is more powerful than that of Nimodipine. Side effects of NBP in behavior was not found. Therefore, NBP seems to be a hopeful drug for the treatment of stroke.

Topics: Animals; Behavior, Animal; Benzofurans; Cerebral Infarction; Dizocilpine Maleate; Male; Neuroprotective Agents; Nimodipine; Rats; Rats, Wistar; Vasodilator Agents

Topics: Animals; Avoidance Learning; Benzofurans; Epilepsy; Female; Hippocampus; Lactones; Memory; Rats; Rats, Inbred Strains

Topics: Animals; Anticonvulsants; Benzofurans; Brain; Epilepsy; Female; Lactones; Rats; Rats, Inbred Strains

Topics: Animals; Anticonvulsants; Benzofurans; China; Electroshock; Female; Male; Mice; Plants, Medicinal; Rats; Structure-Activity Relationship

Topics: Benzofurans; Chemical Phenomena; Chemistry; Plants, Medicinal