phenanthrenes and Disease-Models--Animal

To further evaluate the scope and mechanism of potential cardiotoxicity associated with the antimalarial drug halofantrine, case reports submitted to the US Food and Drug Administration Spontaneous Reporting System were examined. Because halofantrine was associated with electrocardiographic prolongation of the QT interval and ventricular arrhythmias, in vitro cardiac electrophysiologic studies (isolated perfused cardiac model and isolated ventricular myocytes) were conducted to test the hypothesis that halofantrine or its metabolite is responsible for cardiotoxicity.. Although it is difficult to ascertain causality and to estimate overall incidence, a significant number of adverse events related to the cardiovascular system were reported, including QT interval prolongation, life-threatening arrhythmias, and sudden death. The effect of halofantrine and its active metabolite (N-desbutylhalofantrine) on repolarization were examined in an isolated perfused heart model. Results indicate that halofantrine was able to prolong the QT interval, whereas N-desbutylhalofantrine had minimal effect on the QT interval relative to baseline. In an attempt to further elucidate the mechanism of QT interval prolongation, the effects of racemic halofantrine, its stereoisomers, and N-desbutylhalofantrine on repolarizing currents in isolated ventricular myocytes were studied with use of patch-clamp techniques. Halofantrine produced a stereoselective block of the delayed rectifier potassium channel in isolated feline myocytes.. These results indicate that halofantrine is similar to quinidine and class III antiarrhythmics in its ability to prolong repolarization. We conclude that high plasma concentrations of halofantrine should be avoided, especially in women, and that N-desbutylhalofantrine may have potential as a safer antimalarial drug.

Topics: Adolescent; Adult; Adverse Drug Reaction Reporting Systems; Aged; Animals; Antimalarials; Cardiovascular Diseases; Cats; Child; Child, Preschool; Disease Models, Animal; Female; Heart Conduction System; Humans; Infant; Male; Middle Aged; Myocardium; Phenanthrenes

The three-ringed polycyclic aromatic hydrocarbon (PAH) phenanthrene (Phe) has been implicated in the cardiotoxicity of petroleum-based pollution in aquatic systems, where it disrupts the contractile and electrical function of the fish heart. Phe is also found adsorbed to particulate matter and in the gas phase of air pollution, but to date, no studies have investigated the impact of Phe on mammalian cardiac function.. Our objectives were to determine the arrhythmogenic potential of acute Phe exposure on mammalian cardiac function and define the underlying mechanisms to provide insight into the toxicity risk to humans.. Mouse hearts exposed to. To the best of our knowledge, this is the first evidence of direct inhibitory effects of Phe on mammalian cardiac electrical activity at both the whole-heart and cell levels. This electrical dysfunction manifested as an increase in arrhythmia susceptibility due to impairment of both conduction and repolarization. Similar effects in humans could have serious health consequences, warranting greater regulatory attention and toxicological investigation into this ubiquitous PAH pollutant generated from fossil-fuel combustion. https://doi.org/10.1289/EHP12775.

Topics: Action Potentials; Air Pollutants; Animals; Arrhythmias, Cardiac; Disease Models, Animal; Humans; Mammals; Mice; Myocytes, Cardiac; Phenanthrenes; Potassium

Myocardial infarction (MI) leads to pathological cardiac remodeling and heart failure. Sodium tanshinone IIA sulfonate (STS) shows to possess therapeutic potential. The present study aimed to explore the potential role of STS in ventricular remodeling post-MI.. Mice were randomly divided into sham, MI + normal saline (NS) and MI + STS (20.8 mg/kg/day intraperitoneally) groups. MI was established following left anterior descending artery ligation. Cardiac function was evaluated using echocardiography. Scar size and myocardial fibrosis-associated markers were detected using Masson's trichrome staining and western blot analysis (WB). Necrosis and inflammation were assessed using H&E staining, lactate dehydrogenase (LDH) detection, ELISA, immunohistochemical staining, and WB. Furthermore, angiogenesis markers and associated proteins were detected using immunohistochemical staining and WB.. Mice treated with STS exhibited significant improvements in cardiac function, smaller scar size, and low expression levels of α-smooth muscle actin and collagen I and III at 28 days following surgery, compared with the NS-treated group. Moreover, treatment with STS reduced eosinophil necrosis, the infiltration of inflammatory cells, plasma levels of LDH, high mobility group protein B1, interleukin-1β and tumor necrosis factor- α, and protein expression of these cytokines at 3 days. Macrophage infiltration was also decreased in the STS group in the early phase. Additionally, CD31+ vascular density, protein levels of hypoxia-inducible factor- 1α, and vascular endothelial growth factor were elevated in the STS-treated mice at 28 days.. STS improved pathological remodeling post-MI, and the associated therapeutic effects may be a result of a decrease in myocardial necrosis, modulation of inflammation, and an increase in angiogenesis.

Topics: Animals; Cicatrix; Disease Models, Animal; Humans; Inflammation; Mice; Myocardial Infarction; Myocardium; Neovascularization, Pathologic; Phenanthrenes; Vascular Endothelial Growth Factor A; Ventricular Remodeling

Alzheimer's disease (AD) is a most common neurodegenerative disease. Sodium Tanshinone IIA Sulfonate (STS) has been reported to ameliorate AD pathology. However, the underlying mechanism is still unclear. In this study, AD transgenic mouse model (APP/PS1) was used to explore the potential mechanism of STS against AD. Morris water maze and Y-maze tests showed that administration of STS improved learning and memory abilities of APP/PS1 mice. STS reduced the levels of reactive oxygen species and malondialdehyde, while improved the activity of superoxide dismutase in both hippocampus and cortex in APP/PS1 mice. STS inhibited the activity of acetylcholinesterase, while improved the activity of choline acetyltransferase in APP/PS1 mice. In addition, STS elevated the protein expressions of neurotrophic factors and synapse-related proteins in both the hippocampus and cortex in APP/PS1 mice. At last, STS improved the protein expressions of glucose transporter 1 (GLUT1) and low-density lipoprotein receptor-related protein 1 (LRP1). These results indicated that the potential mechanism of STS on AD might be related to Aβ transportation function via GLUT1/LRP1 pathway. HIGHLIGHTS: STS improves cognitive impairment of APP/PS1 mice. STS ameliorates the oxidative stress damage and improves the cholinergic system. STS protects against neuronal dysfunction and enhances the synaptic plasticity. STS mediates the Aβ transportation of BMECs.

Topics: Acetylcholinesterase; Alzheimer Disease; Animals; Cognitive Dysfunction; Disease Models, Animal; Glucose Transporter Type 1; Mice; Mice, Transgenic; Neurodegenerative Diseases; Phenanthrenes

To study the possible mechanism of ghrelin in heart failure and how it works.. In vitro results demonstrated that ghrelin alleviates cardiac function and reduces myocardial fibrosis in rats with heart failure. Moreover, ghrelin intervention increased PTEN expression level and reduced ERK, c-jun, and c-Fos expression level; in vivo experiments demonstrated that ghrelin intervention reduces mast memory expression and increases cardiomyocyte surface area, PTEN expression level, ERK, c-jun, c-Fos expression level, and cell surface area, while ERK blockade suppresses mast gene expression and reduces cell surface area.. In vitro experimental results prove that we have successfully constructed a rat model related to heart failure, and ghrelin can alleviate the heart function of heart failure rats and reduce myocardial fibrosis. In addition, ghrelin is closely related to the decrease of the expression levels of ERK, c-jun, and c-Fos, but it can also increase the expression of PTEN in the rat model; in vivo experiments proved that we successfully constructed an in vitro cardiac hypertrophy model, and the intervention of ghrelin would reduce the expression of hypertrophic memory and increase the surface area of cardiomyocytes, increase the expression level of PTEN, and reduce the expression levels of ERK, c-jun, and c-Fos, while the blockade of PTEN will increase the expression of hypertrophy genes and increase the cell surface area, while the blockade of ERK will increase the expression of hypertrophic genes, which in turn will make the cell surface area reducing.. Ghrelin inhibits the phosphorylation and nuclear entry of ERK by activating PTEN, thereby controlling the transcription of hypertrophic genes, improving myocardial hypertrophy, and enhancing cardiac function.

Topics: Animals; Butadienes; Cell Enlargement; Cell Line; Computational Biology; Disease Models, Animal; Female; Fibrosis; Gene Expression; Ghrelin; Heart Failure; MAP Kinase Signaling System; Mast Cells; Myocytes, Cardiac; Nitriles; Phenanthrenes; PTEN Phosphohydrolase; Rats; Rats, Sprague-Dawley

Triptolide (TP), the main active ingredient of

Topics: Acute Disease; Animals; Antioxidants; Ceruletide; China; Disease Models, Animal; Diterpenes; Epoxy Compounds; Gene Expression Regulation; Hep G2 Cells; Humans; Inflammation; Male; Mice; Mice, Inbred ICR; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Pancreas; Pancreatitis; Phenanthrenes; Reactive Oxygen Species

Topics: Animals; Colitis; Dendritic Cells; Disease Models, Animal; Diterpenes; Epoxy Compounds; Mice; Mice, Inbred C57BL; Phenanthrenes

Alzheimer's disease (AD) is one of the leading causes of dementia. As the first common neurodegenerative disease, there are no effective drugs that can reverse the progression. The present study is to report the anti-AD effect of cryptotanshinone (CTS), a natural product isolated from

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Animals, Genetically Modified; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Disease Models, Animal; Neurodegenerative Diseases; Oxidative Stress; Phenanthrenes; Reactive Oxygen Species

Cerebral ischemia is a major health threat to humankind around the world, and the reperfusion methods may provoke irreversible damages to brain tissues, causing impairment of neurological function. The goal of this study is to investigate the potential neurological protective effect of PJ34, a well-characterized poly (ADP-ribose) polymerase 1 (PARP-1) inhibitor, on cerebral ischemia-reperfusion (I/R)-induced injury of the rat model. The cerebral I/R rats were received (3, 6, or 12 mg/kg) injections of PJ34 or saline at 24 h, 6 h before middle cerebral artery occlusion (MCAO) and 1 h, 24 h, and 48 h after MCAO. All rats were subject to the neurological behavior tests by open field test and Morris water maze test. The expression of pro-inflammatory cytokines, Cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) in cerebral tissues was also determined. Our results demonstrated that the administration of PJ34 dose-dependently ameliorated cerebral I/R-induced injury and improved neurological performance of cerebral I/R rats. We also revealed that PJ34 treatment effectively reduced COX2, iNOS, and pro-inflammatory cytokine levels in the I/R-induced injury tissues. Our finding further supports that inhibition of PARP-1 activity is beneficial for reducing post-I/R-induced brain damage via targeting inflammatory response.

Topics: Animals; Brain; Cognitive Dysfunction; Disease Models, Animal; Dose-Response Relationship, Drug; Humans; Inflammation Mediators; Ischemic Attack, Transient; Male; Morris Water Maze Test; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Reperfusion Injury

Choroidal neovascularization (CNV) is a common cause of irreversible blindness in elderly patients in developed countries, and subretinal fibrosis is an advanced stage of CNV. Currently, there is no effective clinical treatment for subretinal fibrosis.. To investigate whether intravitreal injection of triptolide (TP) could attenuate subretinal fibrosis and determine its underlying mechanisms.. CNV was induced by laser photocoagulation in C57BL/6J mice. Immediately after laser photocoagulation, 1 μl of free TP (10 μg), TP-nanolip-PEG (TP-loaded PEGylated nanoliposomes containing 10 μg TP), or the same volume of phosphate-buffered saline (PBS) was intravitreally administered to each respective group. Areas and ratios of subretinal fibrosis were calculated seven days after laser injury. Additionally, expression levels of M2 macrophage-related markers, molecules of the transforming growth factor (TGF)-β1/Smad signaling pathway, and markers for epithelial-mesenchymal transition (EMT) and endothelial-to-mesenchymal transition (EndoMT) were detected both in vitro and in vivo.. TP could attenuate subretinal fibrosis by suppressing the polarization of M2 macrophages and TGF-β1 induced EMT/EndoMT. TP-nanolip-PEG enhanced the inhibitory effects of TP on subretinal fibrosis, suggesting its therapeutic potential for CNV-related subretinal fibrosis.

Topics: Aged; Animals; Disease Models, Animal; Diterpenes; Epithelial-Mesenchymal Transition; Epoxy Compounds; Fibrosis; Humans; Intravitreal Injections; Lasers; Mice; Mice, Inbred C57BL; Phenanthrenes; Transforming Growth Factor beta1

Peptides from oyster hydrolysate (OPs) have a variety of biological activities. However, its protective effect and exact mechanism on testicular injury remain poorly understood. This study aimed to evaluate the protective effect of OPs on triptolide (TP)-induced testis damage and spermatogenesis dysfunction and investigate its underlying mechanism. In this work, the TP-induced testis injury model was created while OPs were gavaged in mice for 4 weeks. The results showed that OPs significantly improved the sperm count and motility of mice, and alleviated the seminiferous tubule injury. Further study showed that OPs decreased malonaldehyde (MDA) level and increased antioxidant enzyme (SOD and GPH-Px) activities, attenuating oxidative stress and thereby reducing the number of apoptotic cells in the testis. In addition, OPs improved the activities of enzymes (LDH, ALP and ACP) related to energy metabolism in the testis and restored the serum hormone level of mice to normal. Furthermore, OPs promoted the expression of Nrf2 protein, and then increased the expression of antioxidant enzyme regulatory protein (HO-1 and NQO1) in the testis. OPs inhibited JNK phosphorylation and Bcl-2/Bax-mediated apoptosis. In conclusion, OPs have a protective effect on testicular injury and spermatogenesis disorders caused by TP, suggesting the potential protection of OPs on male reproduction.

Topics: Animals; Aquatic Organisms; Crassostrea; Disease Models, Animal; Diterpenes; Epoxy Compounds; Male; Mice; Mice, Inbred ICR; Peptides; Phenanthrenes; Protective Agents; Spermatogenesis; Testis

Our previous study discoveredreactive oxygen species (ROS) and apoptosis inducing factor (AIF) increased after retinal detachment. Parthanatos is a cell death form involving ROS and AIF, which is induced by poly (ADP-ribose) polymerase-1 (PARP-1). Therefore, we investigated whether PJ34 (a PARP-1 inhibitor) could inhibit parthanatos and protect the photoreceptors from cell death after retinal detachment (RD).. Experimental retinal detachment modelswere created in Sprague-Dawley rats by subretinal injection of sodium hyaluronate.PJ34 orDMSO were introduced into subretinal space at RD induction, respectively. The structure of retinas and the morphology of photoreceptors were observed by hematoxylin eosin (H&E) staining and transmission electron microscope (TEM). Parthanatos related proteins (PARP-1, PAR,AIF) were detected by Western blot. The vision-dependent behavior of rat was tested by Morris water maze.. H&E staining and TEM results indicated that the structure and outer nuclear layer (ONL) thickness of retinas were preserved, and the photoreceptors death decreasedwith PJ34 treatment. Western blot showed that the expression of PARP-1, PAR and AIF were decreased withPJ34 treatment. In addition, administration of PJ34 also improved the vision-dependent behavior of rat.. These findings suggested that PJ34 is a potential therapeutic agent that attenuated photoreceptor parthanatos death in retinal detachment through inhibition of PARP-1/AIF pathway.

Topics: Animals; Apoptosis Inducing Factor; Blotting, Western; Cell Survival; Disease Models, Animal; Male; Maze Learning; Microscopy, Electron, Transmission; Parthanatos; Phenanthrenes; Photoreceptor Cells, Vertebrate; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Retinal Detachment

Coronary microembolization (CME) is a prevalent cardiovascular disease, especially nowadays when percutaneous coronary intervention is widely applied. However, neither cardio-protective agents nor devices for distal protection could effectively prevent the occurrence of CME. Therefore, we aimed to develop a new drug for CME. Rats were orally administrated with different doses of Cryptotanshinone (CTS, 5, 15, 45 mg/kg) daily for 2 weeks, respectively, following CME surgery. Then cardiac function and cardiac injury were evaluated in CME rats as well as measuring oxidative stress and apoptosis in cardiomyocytes. Compared to sham group, CME operation induced cardiac dysfunction, cardiac injury, the activation of platelet and endothelium, cardiomyocyte apoptosis and oxidative stress, all of which could be dose-dependently restored by CTS pretreatment. Moreover, NF-κB signaling pathway participated in the development of CME and also in the preventive process of CTS against CME. CTS might serve as a potential and promising candidate drug to prevent the occurrence of CME.

Topics: Administration, Oral; Animals; Apoptosis; Cardiovascular Diseases; Disease Models, Animal; Embolism; Male; Myocytes, Cardiac; NF-kappa B; Oxidative Stress; Phenanthrenes; Rats; Rats, Sprague-Dawley; Signal Transduction

Photoreceptor (PR) dysfunction or death is the key pathological change in retinal degeneration (RD). The death of PRs might be due to a primary change in PRs themselves or secondary to the dysfunction of the retinal pigment epithelium (RPE). Poly(ADP-ribose) polymerase (PARP) was reported to be involved in primary PR death, but whether it plays a role in PR death secondary to RPE dysfunction has not been determined. To clarify this question and develop a new therapeutic approach, we studied the changes in PAR/PARP in the RCS rat, a RD model, and tested the effect of PARP intervention when given alone or in combination with RPE cell transplantation. The results showed that poly(ADP-ribosyl)ation of proteins was increased in PRs undergoing secondary death in RCS rats, and this result was confirmed by the observation of similar changes in sodium iodate (SI)-induced secondary RD in SD rats. The increase in PAR/PARP was highly associated with increased apoptotic PRs and decreased visual function, as represented by lowered b-wave amplitudes on electroretinogram (ERG). Then, as we expected, when the RCS rats were treated with subretinal injection of the PARP inhibitor PJ34, the RD process was delayed. Furthermore, when PJ34 was given simultaneously with subretinal ARPE-19 cell transplantation, the therapeutic effects were significantly improved and lasted longer than those of ARPE-19 or PJ34 treatment alone. These results provide a potential new approach for treating RD.

Topics: Animals; Blotting, Western; Cell Survival; Cell Transplantation; Cells, Cultured; Disease Models, Animal; Electroretinography; In Situ Nick-End Labeling; Phenanthrenes; Photoreceptor Cells, Vertebrate; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Rats; Rats, Mutant Strains; Real-Time Polymerase Chain Reaction; Retinal Degeneration; Retinal Pigment Epithelium

Chinese people have used the root of Salvia miltiorrhiza Bunge (called "Danshen" in Chinese) for centuries as an anticancer agent, anti-inflammatory agent, antioxidant, and cardiovascular disease drug. In addition, Danshen is considered to be a drug that can improve ischemia/reperfusion (I/R)-induced myocardium injury in traditional Chinese medicine. However, Danshen is a mixture that includes various bioactive substances. In this study, we aimed to identify the protective component and mechanism of Danshen on myocardium through network pharmacology and molecular simulation methods. First, cryptotanshinone (CTS) was identified as a potential active compound from Danshen that was associated with apoptosis by a network pharmacology approach. Subsequently, biological experiments validated that CTS inhibited ischemia/reperfusion-induced cardiomyocyte apoptosis in vivo and in vitro. Molecular docking techniques were used to screen key target information. Based on the simulative results, MAPKs were verified as well-connected molecules of CTS. Western blotting assays also demonstrated that CTS could enhance MAPK expression. Furthermore, we demonstrated that inhibition of the MAPK pathway reversed the CTS-mediated effect on cardiomyocyte apoptosis. Altogether, our work screened out CTS from Danshen and demonstrated that it protected cardiomyocytes from apoptosis.

Topics: Animals; Apoptosis; Cells, Cultured; Disease Models, Animal; Drugs, Chinese Herbal; Male; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase 3; Molecular Docking Simulation; Myocardial Reperfusion Injury; Myocytes, Cardiac; Network Pharmacology; Phenanthrenes; Salvia miltiorrhiza; Signal Transduction

Tanshinones, the active ingredients derived from the roots of Salvia miltiorrhiza, have been widely used as traditional medicinal herbs for treating human diseases. Although tanshinones showed anti-inflammatory effects in many studies, large knowledge gaps remain regarding their underlying mechanisms. Here, we identified 15 tanshinones that suppressed the activation of NLRP3 inflammasome and studied their structure-activity relationships. Three tanshinones (tanshinone IIA, isocryptotanshinone, and dihydrotanshinone I) reduced mitochondrial reactive-oxygen species production in lipopolysaccharide (LPS)/nigericin-stimulated macrophages and correlated with altered mitochondrial membrane potentials, mitochondria complexes activities, and adenosine triphosphate and protonated-nicotinamide adenine dinucleotide production. The tanshinones may confer mitochondrial protection by promoting autophagy and the AMP-activated protein kinase pathway. Importantly, our findings demonstrate that dihydrotanshinone I improved the survival of mice with LPS shock and ameliorated inflammatory responses in septic and gouty animals. Our results suggest a potential pharmacological mechanism whereby tanshinones can effectively treat inflammatory diseases, such as septic and gouty inflammation.

Topics: Abietanes; AMP-Activated Protein Kinases; Animals; Autophagy; Disease Models, Animal; Female; Furans; Gout; Humans; Inflammasomes; Inflammation; Male; Mice; Mitochondria; NLR Family, Pyrin Domain-Containing 3 Protein; Phenanthrenes; Quinones; Rats; Reactive Oxygen Species; Shock, Septic; Uric Acid

Alveolar hypercoagulation and fibrinolysis inhibition were associated with the refractory hypoxemia and the high mortality in patient with acute respiratory distress syndrome (ARDS), and NF-κB pathway was confirmed to contribute to the process. Triptolide (TP) significantly inhibited NF-κB pathway and thus depressed accessive inflammatory response in ARDS. We speculate that TP could improve alveolar hypercoagulation and fibrinolytic inhibition in LPS-induced ARDS via NF-κB inactivation.. The aim of this experiment was to explore the efficacy and potential mechanism of TP on alveolar hypercoagulation and fibrinolysis inhibition in LPS-induced ARDS in mice.. 50 μl of LPS (5 mg/ml) was inhalationally given to C57BL/6 mice to set up ARDS model. Male mice were randomly accepted with LPS, LPS + TP (1 μg/kg, 10 μg/kg, 50 μg/kg respectively), or with NEMO Binding domain peptide (NBD), an inhibitor of NF-κB. TP (1 μg/kg, 10 μg/kg, 50 μg/kg) were intraperitoneally injected or 10 μg/50 μl of NBD solution were inhaled 30 min before LPS inhalation. A same volume of normal saline (NS) substituted for TP in mice in control. The endpoint of experiment was at 8 hours after LPS stimulation. Pulmonary tissues were taken for hematoxylin-eosin (HE) staining, wet / dry ratio and for lung injury scores (LIS). Tissue factor (TF) and plasminogen activator inhibitor (PAI)-1 in lung tissue were detected by Western-blotting and by quantitative Real-time PCR(qPCR) respectively. Concentrations of TF, PAI-1, thrombin-antithrombin complex (TAT), procollagen peptide type Ⅲ (PⅢP) and activated protein C (APC) in bronchoalveolar lavage fluid (BALF) were measured by ELISA. NF-κB activation and p65-DNA binding activity in pulmonary tissue were simultaneously determined.. LPS stimulation resulted in pulmonary edema, neutrophils infiltration, obvious alveolar collapse, interstitial congestion, with high LIS, which were all dose-dependently ameliorated by Triptolide. LPS also dramatically promoted the expressions of TF and PAI-1 either in mRNA or in protein in lung tissue, and significantly stimulated the secretions of TF, PAI-1, TAT, PⅢP but inhibited APC production in BALF, which were all reversed by triptolide treatment in dose-dependent manner. TP dose-dependently inhibited the activation of NF-κB pathway induced by LPS, indicated by the changes of phosphorylations of p65 (p-p65), p-IKKα/β and p-IκBα, and weakened p65-DNA binding activity. TP and NBD had same efficacies either on alveolar hypercoagulation and fibrinolysis inhibition or on NF-κB signalling pathway in ARDS mice.. TP dose-dependently improves alveolar hypercoagulation and fibrinolysis inhibition in ARDS mice through inhibiting NF-κB signaling pathway. Our data demonstrate that TP is expected to be an effective selection in ARDS.

Topics: Animals; Disease Models, Animal; Diterpenes; Epoxy Compounds; Fibrinolysis; Lipopolysaccharides; Lung; Lung Injury; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Phenanthrenes; Respiratory Distress Syndrome; Signal Transduction; Thrombophilia; Thromboplastin

Increasing evidence indicates that patients or experimental animals exposure to endotoxin (lipopolysaccharides, LPS) exert deleterious cardiac functions that greatly contribute to morbidity and mortality. The pathophysiologic processes, including NLRP3 inflammasome overactivation and cardiac inflammatory injury, are complicated. Sodium tanshinone IIA sulfonate (STS), a water-soluble derivative of tanshinone IIA, is a naturally occurring compound extracted from Salvia miltiorrhiza and has anti-inflammatory and cardioprotective properties. In this study we examined the effect of STS on endotoxin-induced cardiomyopathy and investigated the underlying mechanisms. An endotoxemic mouse model was established by injecting LPS (10 mg/kg). Different doses of STS were administered intraperitoneally (5, 10, or 50 mg/kg) at different time points (2/12 h, 4/12 h, and 8/12 h) after LPS challenge to assess its effect on survival of mice with endotoxemia. In parallel, cardiac function, myocardial inflammatory cytokines, cardiomyocyte pyroptosis and autophagy were evaluated to determine the extent of myocardial damage due to sepsis in the presence and absence of STS at the optimal dose (10 mg/kg) and time-point (2/12 h). The results demonstrated that STS increased the survival rates, improved the compromised cardiac function and reduced myocardial inflammatory injury associated with enhanced autophagy and mitigated NLRP3 inflammasome activation. Moreover, inhibiting of autophagy or blocking the AMPK pathway reversed STS-elicited prevention of cardiomyopathy and activated the NLRP3 inflammasome in endotoxemic mice. Collectively, our study demonstrates that STS attenuates endotoxemia-induced mortality and cardiomyopathy, which may be associated with promotion of autophagy and inhibition of NLRP3 inflammasome overactivation.

Topics: Animals; Autophagy; Cardiomyopathies; Disease Models, Animal; Echocardiography; Endotoxemia; Endotoxins; Heart Ventricles; Humans; Inflammasomes; Male; Mice; Myocytes, Cardiac; NLR Family, Pyrin Domain-Containing 3 Protein; Phenanthrenes; Pyroptosis

Chronic pancreatitis (CP), characterized by pancreatic fibrosis, is a recurrent, progressive and irreversible disease. Activation of the pancreatic stellate cells (PSCs) is considered a core event in pancreatic fibrosis. In this study, we investigated the role of hydrogen peroxide-inducible clone-5 (Hic-5) in CP. Analysis of the human pancreatic tissue samples revealed that Hic-5 was overexpressed in patients with CP and was extremely low in healthy pancreas. Hic-5 was significant up-regulated in the activated primary PSCs independently from transforming growth factor beta stimulation. CP induced by cerulein injection was ameliorated in Hic-5 knockout (KO) mice, as shown by staining of tissue level. Simultaneously, the activation ability of the primary PSCs from Hic-5 KO mice was significantly attenuated. We also found that the Hic-5 up-regulation by cerulein activated the NF-κB (p65)/IL-6 signalling pathway and regulated the downstream extracellular matrix (ECM) genes such as α-SMA and Col1a1. Therefore, we determined whether suppressing NF-κB/p65 alleviated CP by treating mice with the NF-κB/p65 inhibitor triptolide in the cerulein-induced CP model and found that pancreatic fibrosis was alleviated by NF-κB/p65 inhibition. These findings provide evidence for Hic-5 as a therapeutic target that plays a crucial role in regulating PSCs activation and pancreatic fibrosis.

Topics: Animals; Cells, Cultured; Ceruletide; Cytoskeletal Proteins; Disease Models, Animal; Diterpenes; DNA-Binding Proteins; Down-Regulation; Epoxy Compounds; Fibrosis; Interleukin-6; LIM Domain Proteins; Mice, Knockout; NF-kappa B; Pancreas; Pancreatic Stellate Cells; Pancreatitis, Chronic; Phenanthrenes; Signal Transduction; Transcription Factor RelA; Transforming Growth Factor beta

Exacerbation of chronic obstructive pulmonary disease (COPD) is characterized by acute airway inflammation and mucus hypersecretion, which is by far the most costly aspect of its management. Thus, it is essential to develop therapeutics with low side effects for CODP exacerbation. Sodium tanshinone IIA sulfonate (STS) is a water-soluble derivative of tanshinone IIA isolated as the major active component of Chinese herbal medicine Danshen. Although it possesses anti-inflammatory, anti-oxidative and anti-apoptotic properties, it remains unknown whether STS protects against COPD exacerbation. In this study, we challenged cigarette smoke (CS)-exposed mice with lipopolysaccharide (LPS), and then treated these mice with STS. We found that STS significantly ameliorated pulmonary inflammatory responses, mucus hypersecretion and lung function decline in CS-exposed mice challenged with LPS. STS treatment also significantly attenuated increased IL-6 and IL-8 releases from cigarette smoke extract (CSE)-treated human bronchial epithelial cells (16HBE) challenged with LPS. Mechanistically, STS reduced activation of ERK1/2 and NF-κB in lungs of CS-exposed mice and CSE-treated 16HBE cells challenged with LPS. Taken together, STS protects against acute exacerbation of CS-induced lung injury, which provides a promising and potential therapeutic avenue to halt acute exacerbation of COPD.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents; Cell Line; Cigarette Smoking; Disease Models, Animal; Disease Progression; Drugs, Chinese Herbal; Humans; Lung; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Phenanthrenes; Pulmonary Disease, Chronic Obstructive; Signal Transduction

Cisplatin is a well-known chemotherapy medication used to treat numerous cancers. However, treatment with cisplatin in cancer therapy has major side effects, such as nephrotoxic acute kidney injury. Adult vertebrate kidneys are commonly used as models of cisplatin-induced nephrotoxic acute kidney injury. Embryonic zebrafish kidney is more simplified and is composed simply of two nephrons and thus is an excellent model for the investigation of cisplatin nephrotoxicity. Here, we developed a novel model to induce cisplatin nephrotoxicity in adult zebrafish and demonstrated that intraperitoneal injection of cisplatin caused a decline in kidney proximal tubular function based on fluorescein-labeled dextran uptake and alkaline phosphatase staining. We also showed that cisplatin induced histological injury of the kidney tubules, quantified by tubular injury scores on the periodic acid-Schiff-stained kidney sections. As shown in a mouse model of cisplatin-induced nephrotoxicity, the activation of poly(ADP-ribose) polymerase (PARP), an enzyme implicated in cisplatin-induced cell death, was markedly increased after cisplatin injection in adult zebrafish. Furthermore, pharmacological inhibition of PARP using a specific PARP inhibitor PJ 34 hydrochloride (PJ34) or 3-aminobenzamide ameliorated kidney proximal tubular functional and histological damages in cisplatin-injected adult zebrafish kidneys. Administration of a combination of PARP inhibitors PJ34 and 3-aminobenzamide additively protected renal function and histology in zebrafish and mouse models of cisplatin nephrotoxicity. In conclusion, these data suggest that adult zebrafish are not only suitable for drug screening and genetic manipulation but also useful as a simplified but powerful model to study the pathophysiology of cisplatin nephrotoxicity and establish new therapies for treating human kidney diseases.

Topics: Animals; Benzamides; Cisplatin; Disease Models, Animal; DNA Damage; Kidney Diseases; Kidney Tubules; Male; Mice, Inbred C57BL; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Signal Transduction; Zebrafish; Zebrafish Proteins

Idiopathic pulmonary fibrosis (IPF) is a fibrotic interstitial pneumonia that causes pulmonary tissue damage and functional impairment. To investigate the effects of cryptotanshinone on pulmonary fibrosis, the expression of NIH/3T3, HPF, and rat primary pulmonary fibroblasts was measured and found to be inhibited by CPT in a time- and concentration-dependent manner, and the upregulation of α-SMA expression in NIH/3T3 and HPF cells, which had been stimulated by TGFβ-1, was decreased after CPT administration. We observed that CPT could reverse the increase in α-SMA expression and vimentin and the decrease in E-cad expression in A549 cells, which had been induced by 5 ng/mL TGFβ-1, indicating that CPT has inhibitory effects in the EMT process. A BLM-induced pulmonary fibrosis model was established in C57BL/6 mice. The lung coefficient and hydroxyproline content increased significantly in the BLM-induced group and were decreased in the CPT-treated group. The expression levels of collagen-I and α-SMA and the phosphorylation level of Stat3 were significantly increased, and CPT treatment decreased these levels. Furthermore, the results from the flow cytometry analysis indicated that, in lung tissues, the frequencies of MDSCs, macrophages, DCs and T cells were considerably increased in the BLM-induced group, while CPT treatment reduced these immunocyte populations.

Topics: Animals; Antibiotics, Antineoplastic; Bleomycin; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial-Mesenchymal Transition; Humans; Male; Mice; Mice, Inbred C57BL; Phenanthrenes; Pulmonary Fibrosis; Rats

The incidence of ulcerative colitis (UC) is increasing in recent years. The protective effect of cryptotanshinone, a natural compound from Salvia miltiorrhiza Bunge, on UC was investigated both in vivo and in vitro models. UC model was established by dextran sulfate sodium administration in drinking water and cryptotanshinone was orally administrated. RAW264.7 cells were stimulated by lipopolysaccharide (LPS) with or without cryptotanshinone pretreatment. The body weights and disease activity index (DAI) were recorded. The pathological alterations were evaluated by H&E staining. The levels of pro-inflammatory cytokines in colon tissues and cell culture medium were determined with enzyme-linked immune sorbent assay (ELISA) kits. The protein expression was detected by Western blotting and immunohistochemistry. Results showed that cryptotanshinone significantly increased the body weight and colon length, reduced the score of DAI, and improved pathological changes. Furthermore, the expression of inducible nitric oxide synthase, cyclooxygenase-2, receptor-interacting protein kinase 3, NF-κB p65 and the secretion of tumor necrosis factor-α, IL-6 in colon tissues and LPS-stimulated cells were significantly inhibited by cryptotanshinone. Besides, cryptotanshinone significantly inhibited LPS-triggered toll-like receptor 4 luciferase reporter activity with an IC

Topics: Animals; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Drugs, Chinese Herbal; Inflammation; Male; Mice; Phenanthrenes

Chemotherapy-induced intestinal mucositis (CIM) is a principal reason for reduced living quality of patients undergoing chemotherapy. Growing evidence showed gut microbiota played an important role in the development of intestinal mucositis. Dihydrotanshinone I (DHTS) is a liposoluble extract of Salvia miltiorrhiza Bunge with many bioactivities. Here we investigated the effect of DHTS on intestinal mucositis induced by 5-fluorouracil and irinotecan in mice. We detected the degree of intestinal mucosal damage and inflammatory response in CIM mice with or without DHTS administration. The body weight and disease activity index (DAI) of mice were monitored each day. H&E staining was used to evaluate pathological damage. The contents of interleukin 6 (IL-6), tumor necrosis factor (TNFα), diacylglycerol (DAO) and triglyceride (TG) in serum were determined by commercial kits. We also investigated the changes of fecal microbiota by 16S rRNA high-throughput sequencing. Spearman correlation analysis was used to evaluate the correlation between fecal microbiota and inflammatory factors. Tax4Funwas performed to infer the potential function of the microbial community. Results showed DHTS significantly reduced DAI, intestinal mucosal damage and inflammatory response in CIM mice by decreasing serum IL-6 and TNFα. In addition, there is an intense correlation between fecal microbiota and inflammatory factors. DHTS efficiently reversed disordered fecal microflora close to normal and increased the abundance of g__Akkermansia. DHTS also enriched bacterial species which promote butyric acid metabolism or negatively correlated with inflammatory factors. Besides, species enriched by DHTS in fecal microbiota were probably involved in glutamine production and ammonia oxidation. In conclusion, our study provides evidence that DHTS effectively attenuates CIM induced by 5-fluorouracil and irinotecan in mice. Regulation of the composition and function of fecal microbiota probably plays a critical role in the therapeutic effect of DHTS in CIM mice.

Topics: Animals; Anti-Inflammatory Agents; Antineoplastic Agents; Bacteria; Colon; Diglycerides; Disease Models, Animal; Feces; Fluorouracil; Furans; Gastrointestinal Microbiome; Interleukin-6; Intestinal Mucosa; Irinotecan; Male; Mice, Inbred C57BL; Mucositis; Phenanthrenes; Quinones; Triglycerides; Tumor Necrosis Factor-alpha

The cardiotoxicity of doxorubicin (DOX) reduces the quality of life and prognosis of cancer patients, and therefore its clinical application has been largely restricted. This study aimed to assess the effects of cryptotanshione (CPT) on DOX-induced rat cardiac insufficiency.. CPT treatment significantly suppressed apoptosis. Transcriptomic profiling and bioinformatics analysis can be used to evaluate the cardio-protective effect of CPT through inactivating p53 signaling pathway in the doxorubicin-mediated myocardial damage model.. F-actin staining and flow cytometry were used to assess the effects of CPT on cardiomyocytes.

Topics: Animals; Apoptosis; Cardiotonic Agents; Cardiotoxicity; Computational Biology; Databases, Genetic; Disease Models, Animal; Doxorubicin; Gene Expression Profiling; Heart; Heart Diseases; Metabolic Networks and Pathways; Myocytes, Cardiac; Phenanthrenes; Rats; Reactive Oxygen Species; Signal Transduction; Transcriptome

Sodium tanshinone IIA sulfonate (STS) can protect against brain damage induced by stroke. However, the neural protection mechanism of STS remains unclear. We investigated whether STS performs its protective function by suppressing autophagy and inflammatory activity during brain injury. We established a transient middle cerebral artery occlusion and reperfusion (MCAO/R) model by blocking the left middle cerebral artery with a thread inserted through the internal carotid artery for 1 h, followed by reperfusion for 48 h either with or without STS and the autophagy inhibitor 3-methyladenine (3-MA). Neuroprotective effects were determined by evaluating infarction, brain edema, and neurological deficits. The numbers of microglia-derived macrophages, monocyte-derived microglia, T cells, and B cells in the brains were measured, based on the surface marker analyses of CD45, CD11b, B220, CD3, and CD4 using fluorescence-assisted cell sorting. STS (10, 20, 40 mg/kg) was able to significantly reduce infarct volumes, improve neurological deficits, and reduce brain water contents. STS treatment reduced neuroinflammation, as assessed by the infiltration of macrophages and neutrophils, corresponding with reduced numbers of macrophages, T cells, and B cells in ischemia/reperfusion (I/R) brains. In addition, STS treatment also attenuated the upregulation of autophagy associated proteins, such as LC3-II, Beclin-1 and Sirt 6, which was induced by MCAO. These results demonstrated that STS can provide remarkable protection against ischemic stroke, possibly via the inhibition of autophagy and inflammatory activity.

Topics: Animals; Autophagy; Brain Ischemia; Disease Models, Animal; Infarction, Middle Cerebral Artery; Inflammation; Neuroprotective Agents; Phenanthrenes; Rats; Rats, Sprague-Dawley; Reperfusion Injury

Triptolide (TP), as the main component of Tripterygium Wilfordii (TW), can induce obvious liver injury when exerting the therapeutic effect. However, in our previous study, Catalpol (CAT), the main active ingredient of Rehmannia Glutinosa (RG), was shown to increase the drug clearance rate of TP and to attenuate TP-induced hepatotoxicity. Thus the present study aims to address the roles of phase I and II metabolic enzymes and the nuclear receptors in the detoxification process of TP, to analyze the mechanism of CAT reducing hepatotoxicity. For this purpose, SD rats and human liver cell line L-02 and HepG2 cells were selected, and treated with TP or the combination of TP and CAT in our study. Then the effect of CAT on detoxification of TP was analyzed, and the roles of phase I metabolic enzymes cytochrome P450 3A2/4 (CYP3A2/4) and phase II metabolic enzyme UDP-glucuronosyltransferase 1A6 (UGT1A6) and their related nuclear receptor regulations were evaluated. It was found that TP inhibited the transcription of CYP3A2/4. And through the constitutive androstane receptor (CAR) pathway, CAT not only significantly changed this inhibition and increased the expression of CYP3A2/4 but also increased the expression of CYP2C9, both of which are phase I detoxification enzymes of TP. And with the gene-silenced experiment, it was confirmed that this regulation was CAR-dependent. We also found that CAT could continue to exert a certain protective effect after CAR was silenced, with UGT1A6, the phase II detoxification enzyme of TP, significantly induced. And this was closely related to the enhanced transcriptional regulation of the nuclear factor erythroid 2-related factor 2 (NRF2) pathway. In conclusion, our results reveal that CAT can induce TP's phase I detoxification enzymes CYP3A2/4 and CYP2C9 through the CAR pathway, and induce TP's phase II detoxification enzyme UGT1A6 via the NRF2 pathway when CAR is strongly inhibited. And this coordinate regulation of CAT may be an important source of the effect for CAT to increase TP metabolic conversion and reduce TP hepatotoxicity.

Topics: Animals; Chemical and Drug Induced Liver Injury; Constitutive Androstane Receptor; Cytochrome P-450 Enzyme System; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Glucuronosyltransferase; Hep G2 Cells; Hepatocytes; Humans; Iridoid Glucosides; Liver; Metabolic Detoxication, Phase I; Metabolic Detoxication, Phase II; NF-E2-Related Factor 2; Phenanthrenes; Rats, Sprague-Dawley; Receptors, Cytoplasmic and Nuclear; Signal Transduction

Gestational diabetes mellitus (GDM) is a conditional diabetes which is defined as any degree of glucose intolerance or high blood glucose levels during any phase of pregnancy. It causes chronic severe damage to health of the pregnant women and their offspring. In this study, we aimed to study the protective effects of Cryptotanshinone on GDM-related impairments. We measured blood glucose levels, serum insulin levels, biochemical indexes, oxidative stress, inflammation and the activation of NF-κB signaling pathway in the blood and placenta of GDM mice. It is found that Cryptotanshinone significantly decreased blood glucose levels, oxidative stress, inflammation and NF-κB signaling with an increase of serum insulin levels in the placenta and blood of GDM mice. Taken together, Cryptotanshinone effectively ameliorated GDM, which suggested that Cryptotanshinone could be served as a promising therapeutic drug for GDM patients.

Topics: Administration, Oral; Animals; Diabetes, Gestational; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Female; Inflammation; Mice; Mice, Inbred C57BL; Oxidative Stress; Phenanthrenes; Pregnancy

Silicosis is characterized by pulmonary fibrosis due to long-term inhalation of silica particles. Although the cause of this serious disease is known, its pathogenesis remains unclear and there are currently no specific treatments. Recent studies have shown that the anti-oxidant transcription factor Nrf2 is expressed at reduced levels in fibrotic foci, which may be related to disease progression. However, the molecular mechanisms by which this might occur have yet to be elucidated. Sodium tanshinone IIA sulfonate (STS), an extract of Salvia miltiorrhiza, is used in traditional Chinese medicine in the treatment of coronary heart disease. STS has been shown to play a strong anti-oxidative role in various organs. Here, we employed a rat model to explore the effects of STS on oxidative stress and the progression of fibrosis in silicosis. STS significantly reduced collagen deposition in the lungs, thereby antagonising silicosis. Immunohistochemical and immunofluorescence staining showed that Nrf2 was differentially expressed in lung cells during silica induced fibrosis, and chromatin immunoprecipitation-sequencing experiments demonstrated that Nrf2 promoted the expression of the antioxidant proteins thioredoxin and thioredoxin reductase. Our results suggest that the anti-fibrotic effects of STS may be related to upregulation of Nrf2 nuclear expression, especially in fibrotic lesions, and the promotion of thioredoxin and thioredoxin reductase expression. Our findings may open up new avenues for the development of STS as a treatment for silicosis.

Topics: A549 Cells; Animals; Disease Models, Animal; Drugs, Chinese Herbal; Humans; Inhalation Exposure; Male; Mice; NF-E2-Related Factor 2; Particle Size; Phenanthrenes; Pulmonary Fibrosis; Rats; Rats, Wistar; RAW 264.7 Cells; Silicon Dioxide; Silicosis; Surface Properties; Thioredoxins

Triptolide (TP) exhibits effective activity against colon cancer in multiple preclinical models, but the mechanisms underlying the observed effects are not fully understood. Sphingosine-1-phosphate (S1P) is a potent bioactive sphingolipid involved in the regulation of colon cancer progression. The aim of this study was to investigate the effect of TP on the sphingosine kinase (SPHK)-S1P signaling pathway in colitis-associated colon cancer.. An azoxymethane (AOM)/dextran sulfate sodium (DSS) mouse model and the THP-1 cell line were used to evaluate the therapeutic effects and mechanisms of TP in colitis-associated colon cancer (CACC). Various molecular cell biology experiments, including Western blotting, real-time PCR and immunofluorescence, were used to obtain relevant experimental data. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was also established to detect the levels of S1P in tissue and plasma.. In the AOM/DSS mouse model, TP treatment induced a dose-dependent decrease in tumor incidence and inhibited macrophage recruitment and M2 polarization in the tumors. TP also efficiently decreased the S1P levels and SPHK1/S1PR1/S1PR2 expression and significantly inhibited activation of the S1P-mediated phosphorylation of ERK protein in macrophages.. The results indicated that TP might influence the recruitment and polarization of tumor-associated macrophages by suppressing the SPHK-S1P signaling pathway.

Topics: Animals; Azoxymethane; Colitis; Colitis-Associated Neoplasms; Colon; Dextran Sulfate; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Humans; Lysophospholipids; Male; Mice, Inbred BALB C; Mice, Inbred ICR; Mice, Nude; Phenanthrenes; Phosphotransferases (Alcohol Group Acceptor); Signal Transduction; Sphingosine; THP-1 Cells; Tumor-Associated Macrophages

Dihydrotanshinone I (DHTS), extracted from Salvia miltiorrhiza, was found to be the most effective compound of tanshen extracts against cancer cells in our previous studies. However, the therapeutic benefits and underlying mechanisms of DHTS on ovarian cancer remain uncertain. In this study, we demonstrated the cytocidal effects of DHTS on chemosensitive ovarian cancer cells with or without platinum-based chemotherapy. DHTS was able to inhibit proliferation and migration of ovarian cancer cells in vitro and in vivo through modulation of the PI3K/AKT signalling pathways. Combinatorial treatment of DHTS and cisplatin exhibited enhanced DNA damage in ovarian cancer cells. Overall, these findings suggest that DHTS induces ovarian cancer cells death via induction of DNA damage and inhibits ovarian cancer cell proliferation and migration.

Topics: Animals; Carcinoma, Ovarian Epithelial; Cell Death; Cell Line, Tumor; Cell Movement; Cell Proliferation; Class I Phosphatidylinositol 3-Kinases; Disease Models, Animal; Drug Resistance, Neoplasm; Female; Furans; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Neoplasm Invasiveness; Neoplasm Metastasis; Ovarian Neoplasms; Phenanthrenes; Phosphatidylinositol 3-Kinases; Platinum; Proto-Oncogene Proteins c-akt; Quinones; Signal Transduction; Transcription, Genetic; Zebrafish

Cryptotanshinone (CRY) has been demonstrated to reverse reproductive disorders. However, whether CRY is effective in the treatment of polycystic ovary syndrome (PCOS) remains unknown. The aim of the present study was to evaluate the therapeutic potential of CRY in PCOS. A rat model of PCOS was established by daily injection of human chorionic gonadotropin and insulin for 22 days. Total body weight and ovarian weight, as well as the levels of luteinizing hormone (LH) and the LH to follicle‑stimulating hormone (FSH) ratio (LH/FSH) significantly increased in rats with PCOS, compared with controls. Moreover, the levels of testosterone (T), tumor necrosis factor (TNF)‑α and high‑mobility group box 1 protein (HMGB1) also increased. However, CRY treatment attenuated the increase in body weight, ovarian weight, LH, LH/FSH ratio, T, TNF‑α and HMGB1 levels, compared with the PCOS group. Treatment with CRY also reduced NF‑κB/p65, HMGB1 and toll‑like receptor (TLR)4 mRNA and protein expression levels in the ovarian tissue and granulosa cells, both in vitro and in vivo. Thus, CRY significantly mitigated the changes in body weight, ovary weight, hormone levels and inflammatory factor levels observed in rats with PCOS. Thus, CRY protects against PCOS‑induced damage of ovarian tissue, possibly through a regulatory pathway involving HMGB1, TLR4 and NF‑κB.

Topics: Animals; Body Weight; Cell Proliferation; Cell Survival; Disease Models, Animal; Female; Gene Expression Regulation; HMGB1 Protein; NF-kappa B; Organ Size; Phenanthrenes; Polycystic Ovary Syndrome; Rats; Signal Transduction; Toll-Like Receptor 4

Triptolide (TP), a principal bioactive component of traditional Chinese medicine Tripterygium wilfordii Hook. F., has been shown to have immunosuppressive/anti-inflammatory actions in vitro. Moreover, it is well established that inflammatory mechanisms contribute to the progression of hypertension-induced renal injury. Therefore, this study was performed to determine the protective effects of TP on renal injury in salt-sensitive hypertension and to identify the possible mechanisms for TP-induced protection.. Ten-week-old male C57BL/6 mice were subjected to uninephrectomy and deoxycorticosterone acetate (DOCA)-salt treatment with or without intraperitoneal administration of various concentrations of TP.. Five weeks after the treatment, systolic blood pressure measured by tail-cuff plethysmography increased in DOCA-salt-treated mice, but no difference was found between DOCA-salt-treated mice with or without TP treatment. Treatment with TP dose-dependently attenuated increments in urinary albumin and 8-isoprostane excretion, and glomerulosclerosis and tubulointerstitial injury and fibrosis in DOCA-salt-treated mice. Moreover, our data showed that treatment with TP dose-dependently inhibited DOCA-salt-induced interstitial monocyte/macrophage infiltration associated with decreases in renal levels of proinflammatory cytokine/chemokine and adhesion molecule, as well as renal activated NF-κB concentrations. Our results also demonstrated that suppression of inflammatory responses with dexamethasone, an immunosuppressive agent, alleviated DOCA-salt hypertension-induced renal injury.. TP treatment induced renal protection associated with inhibition of monocyte/macrophage-mediated inflammatory responses without lowering blood pressure. Thus, our data for the first time indicate that TP treatment ameliorates renal injury possibly via attenuating inflammatory responses in salt-sensitive hypertension.

Topics: Animals; Anti-Inflammatory Agents; Cell Adhesion Molecules; Cytokines; Desoxycorticosterone Acetate; Disease Models, Animal; Diterpenes; Epoxy Compounds; Hypertension; Inflammation Mediators; Kidney; Kidney Diseases; Male; Mice, Inbred C57BL; Nephrectomy; NF-kappa B; Phenanthrenes; Signal Transduction; Sodium Chloride, Dietary

Inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn's disease (CD), is a group of chronic and incurable inflammatory diseases involving the gastrointestinal tract. In this study, we investigated the anti-inflammatory effects of triptolide in a dextran sulfate sodium (DSS)-induced mouse colitis model and LPS-activated macrophages and explored the specific molecular mechanism(s). In mice, triptolide treatment showed significant relief and protection against colitis, and it markedly reduced the inflammatory responses of human monocytes and mouse macrophages. Pharmacological analysis and weighted gene co-expression network analysis (WGCNA) suggested that PDE4B may be an important potential targeting molecule for IBD. Exploration of the specific mechanism of action indicated that triptolide reduced the production of ROS, inhibited macrophage infiltration and M1-type polarization by activating the NRF2/HO-1 signaling pathway, and inhibited the PDE4B/AKT/NF-

Topics: Animals; Anti-Inflammatory Agents; Biomarkers; Colitis; Computational Biology; Cytokines; Dextran Sulfate; Disease Models, Animal; Diterpenes; Epoxy Compounds; Gene Expression Profiling; Inflammation Mediators; Inflammatory Bowel Diseases; Lipopolysaccharides; Macrophage Activation; Macrophages; Male; Mice; Models, Biological; Phenanthrenes; RAW 264.7 Cells; Reactive Oxygen Species; Signal Transduction

Diabetes is a strong risk factor of peripheral arterial disease (PAD), and also leads to impaired perfusion recovery in the ischemic limb, which eventually results in poor outcomes in PAD patients. Sodium Tanshinone IIA Sulfonate (STS), a monomer from herbs, has been shown to improve the outcomes in a variety of ischemic disease including myocardial infarction. However, the effects of STS treatment in PAD is not known.. Unilateral femoral artery was ligated in mice as experimental PAD models, STS treatment improved perfusion recovery, increased capillary densities, decreased reactive oxygen species (ROS) level and microRNA-133a (miR-133a) expression in the ischemic hindlimb in diabetic mice; however, STS did not change perfusion recovery in non-diabetic C57BL/6 mice. Ischemic muscle tissue from diabetic mice was harvested 7 days after femoral ligation for biochemical test, STS resulted in reduced malondialdehyde (MDA), and increased GTP cyclohydrolase 1 (GCH1) and cyclic guanine monophosphate (cGMP) levels. In addition, STS treatment increased miR-133a expression in endothelial cells isolated from ischemic muscle tissue of diabetic mice. In endothelial cells cultured in high glucose medium, STS increased tube formation and nitric oxide (NO) production, and reduced cellular ROS level and miR-133a expression under simulated ischemic condition. In addition, GCH1 inhibitor or miR-133a overexpression using exogenous microRNA mimic blunted STS-induced angiogenic effects and ROS neutralization in cultured endothelial cells under hyperglycemic and hypoxic conditions.. These findings demonstrate STS improves angiogenesis via inhibiting miR-133a expression and increasing GCH-1 protein levels in experimental PAD with diabetes.

Topics: Animals; Diabetes Mellitus, Experimental; Disease Models, Animal; Drugs, Chinese Herbal; Hindlimb; Human Umbilical Vein Endothelial Cells; Humans; Hyperglycemia; Ischemia; Male; Mice; Mice, Inbred C57BL; MicroRNAs; Neovascularization, Physiologic; Peripheral Arterial Disease; Phenanthrenes; Phytotherapy; Reactive Oxygen Species; Salvia miltiorrhiza

The aim of this study was to investigate the effects of triptolide on the tooth movement and root resorption in rats during orthodontic treatment.. A total of 48 male Wistar rats were divided into three groups of 16 each. The right maxillary first molars of rats were drawn mesially by closed coil nickel-titanium spring with a force of 50 g. The two experimental groups received intraperitoneal injections of triptolide for 14 days at a dose of 15 µg/kg/day and 30 µg/kg/day, respectively. The control group received vehicle injections. After 14 days, the rats were humanely killed. The amount of tooth movement was measured. Eight rats from each group were randomly chosen for analysis of the percentage of root resorption area by scanning electron microscopy. For the remaining eight rats in each group, the H&E staining, tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemistry analysis were performed.. The amount of tooth movement and the ratio of root resorption area were significantly decreased in the triptolide-treated rats. The number of TRAP-positive cells was significantly lower in triptolide-treated groups. Moreover, the expression of nuclear factor kappa B ligand (RANKL) was reduced. In contrast, the expression of osteoprotegerin was significantly up-regulated. In the tension side, the expressions of runt-related transcription factor 2 and osteocalcin were significantly enhanced by triptolide injection.. Triptolide injection could arrest orthodontic tooth movement and reduce root resorption in rats via inhibition of osteoclastogenesis. In addition, triptolide may exert a positive effect on osteoblastogenesis.

Topics: Administration, Oral; Animals; Disease Models, Animal; Diterpenes; Epoxy Compounds; Male; Orthodontic Appliances, Removable; Phenanthrenes; Rats; Rats, Wistar; Root Resorption; Tooth Movement Techniques

Triptolide has been widely reported to exhibit potential therapeutic value in multiple inflammatory diseases, such as rheumatoid arthritis, systemic lupus erythematosus, and psoriasis. Although its safety and efficacy as an anti-inflammatory agent have been verified by many studies, the effect of triptolide on osteoarthritis (OA) was not clearly understood. In this study, we found that triptolide prevented OA development in a surgical destabilization of the medial meniscus (DMM) mouse model. In addition, triptolide inhibited both DMM-induced and LPS-induced expression of pro-inflammatory cytokines in the human monocytic cell line THP-1. Further mechanistic studies showed that the reduction of pro-inflammatory cytokines by triptolide was mediated by the upregulation of nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3 (NLRP3) and downregulation of caspase-1. Finally, we identified that hsa-miR-20b, a microRNA targeting the NLRP3 gene, was downregulated by triptolide. This study provides a novel insight into the effect on triptolide in preventing OA pathogenesis.

Topics: Animals; Cell Line; Cytokines; Disease Models, Animal; Diterpenes; Down-Regulation; Epoxy Compounds; Humans; Male; Mice; Mice, Inbred C57BL; MicroRNAs; Monocytes; NLR Family, Pyrin Domain-Containing 3 Protein; Osteoarthritis; Phenanthrenes; THP-1 Cells; Up-Regulation

The objective of this present study was to develop and evaluate the triptolide-loaded nanostructured lipid carriers (TPL-NLCs) for transdermal drug delivery system (TDDS). TPL-NLCs was prepared with emulsification technique and optimized by central composite design of a response surface methodology (CCD-RSM). The optimized TPL-NLCs were spherical and physically stable with the average size of 139.6.0 ± 2.53 nm and Zeta potential of -36.03 ± 2.41 mV. The encapsulation efficiency and drug loading were 97.15% ± 9.46 and 10.35% ± 1.12, respectively. Moreover, the in vitro release study showed that TPL-NLCs had a sustained release profiles and the in vitro penetration study indicated that TPL-NLCs could effectively penetrate into skin. Besides, the methodology of skin-blood synchronous microdialysis was established to evaluate the pharmacokinetics of TPL-NLCs in vivo and the results displayed that the TPL concentration in skin was higher than that in blood. And TPL-NLCs presented a remarkable effect of decreasing knee edema, inhibiting inflammation by regulating the levels of TNF-α, IL-1β and IL-6, which indicated that TPL-NLCs was a promising topical administration in treatment of edema and inflammation associated with rheumatoid arthritis (RA).

Topics: Administration, Cutaneous; Animals; Arthritis, Rheumatoid; Delayed-Action Preparations; Disease Models, Animal; Diterpenes; Drug Carriers; Drug Delivery Systems; Drug Liberation; Edema; Epoxy Compounds; Immunosuppressive Agents; Lipids; Male; Microdialysis; Nanostructures; Particle Size; Phenanthrenes; Rats; Rats, Sprague-Dawley; Skin Absorption

The strong human immunity and the associated toxicities of attenuated Salmonella severely limit the clinical use of Salmonella in tumour suppression. In the present study, we constructed an engineered VNP20009-DNase I strain and evaluated the synergistic effects of triptolide (TPL) and VNP20009-DNase I against melanoma in mice. Our results indicated that TPL could significantly inhibit the cell growth and cell migration and significantly enhanced the apoptosis rate of B16F10 cells in vitro. The in vivo results indicated that TPL markedly improved tumour colonisation of VNP20009-DNase I and led to a larger necrotic area in the melanoma. Moreover, the combination therapy significantly suppressed tumour volume and prolonged the life span of mice (P < 0.05) by upregulating the expression of Bcl-2/Bax and Caspase-3 and by downregulating the TLR4/NF-κB signalling, the expression of p-AKT/AKT and the production of proinflammatory factors. Therefore, the sound synergistic anti-tumour effects of TPL and VNP20009-DNase I indicate that the unconventional application of TPL and biological agents, approved by the China Food and Drug Administration (CFDA), can result in improved anti-cancer therapeutic outcomes.

Topics: Animals; Antineoplastic Agents, Alkylating; Cell Line, Tumor; Cell Proliferation; Combined Modality Therapy; Deoxyribonuclease I; Disease Models, Animal; Diterpenes; Drug Carriers; Epoxy Compounds; Genetic Vectors; Humans; Melanoma; Mice; Models, Biological; Phenanthrenes; Plasmids; Salmonella; Salmonella Infections; Treatment Outcome; Vaccines, DNA

The purpose of this study was to prepare a novel cryptotanshinone-loaded nanoemulsion (Cry LN) and to evaluate its prevention effect on the postoperative peritoneal adhesions (PPA). The Cry LN was prepared by high-pressure homogenization method, and various methods were used to investigate the physicochemical properties. The results showed that Cry LN has nanoscale particle size with uniform distribution and could slowly release the incorporated drug compared with Cry solution. With superior safety, Cry LN could increase the ratio of tissue plasminogen activator (tPA)/plasminogen activator inhibitor-1 (PAI-1) up to 673% compared with control group. Furthermore, in vivo animal study confirmed the Cry LN activated the fibrinolytic system and successfully prevented PPA formation in rat. In conclusion, Cry nanoemulsion could be considered as a potentially promising and effective strategy for PPA treatment.

Topics: Animals; Cell Line; Disease Models, Animal; Emulsions; Fibrinolysis; Humans; Male; Mice; Mice, Inbred BALB C; Nanoparticles; Peritoneal Diseases; Phenanthrenes; Plasminogen Activator Inhibitor 1; Postoperative Complications; Rats; Rats, Sprague-Dawley; Surgical Procedures, Operative; Tissue Adhesions; Tissue Plasminogen Activator

Triptolide is a biologically active component of the Chinese antirheumatic herbal remedy Tripterygium wilfordii Hook F, which has been shown to be effective in treating murine lupus. However, its immunological mechanisms are poorly understood. Regulatory T cells (Treg) are pivotal for maintaining peripheral self-tolerance and controlling autoimmunity. This study was undertaken to examine the therapeutic effect of triptolide in lupus mice and the related molecular mechanisms. Our results showed that triptolide treatment ameliorated serum anti-dsDNA, proteinuria and renal histopathologic assessment in MRL/lpr mice, induced the miR-125a-5p expression and enhanced the proportion of Treg in vivo. In vitro, triptolide upregulated the proportion of Treg and the miR-125a-5p expression. Down-regulation of the miR-125a-5p expression reversed the effect of triptolide on Treg. In conclusion, triptolide could induce Treg and the miR-125a-5p expression in vivo and in vitro. Inhibiting the effect of miR-125a-5p could counteract the effect of triptolide on inducing Treg. The study has strong implications for the therapeutic applications of triptolide in systemic lupus erythematosus.

Topics: Animals; Antibodies, Antinuclear; Antirheumatic Agents; Cell Proliferation; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Humans; Kidney; Lupus Erythematosus, Systemic; Mice; Mice, Inbred C57BL; Mice, Inbred MRL lpr; MicroRNAs; Phenanthrenes; Proteinuria; T-Lymphocytes, Regulatory; Up-Regulation

Peripheral nerve injuries often induce neuropathic pain through inflammation. Cryptotanshinone isolated from Salvia miltiorrhiza Bunge has been found to exert anti-inflammatory and analgesic activities. Thus, this study aimed to determine whether cryptotanshinone inhibits chronic constriction injury (CCI)-induced neuropathic pain in rats and its mechanism of action. CCI was performed by applying four loose ligatures to rat sciatic nerve. Cryptotanshinone was orally administered using pre-surgery, acute or repeated post-surgery treatment. The pain behaviors were determined by recording paw withdrawal mechanical threshold (PWMT) and thermal withdrawal latency (TWL). ELISA kits were used to measure interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α levels. qRT-PCR were performed to detect IL-6, IL-1β, TNF-α, PI3K and Akt expression. The phosphorylation of PI3K/Akt signaling was assessed using western blotting. PWMT and TWL in CCI group were higher than those in the control and sham groups. The acute post-CCI cryptotanshinone treatment but not pre-surgery treatment reduced PWMT and TWL. The effect of cryptotanshinone is more prominent when it was repeatedly administered after CCI. The CCI-induced increase in IL-6, IL-1β, TNF-α, PI3K/Akt signaling and their phosphorylation was also suppressed by repeated post-CCI cryptotanshinone treatment. This study suggested that post-CCI cryptotanshinone treatment reduced the surgery-induced neuropathic pain by suppressing PI3K/Akt signaling therefore inhibited inflammation.

Topics: Analgesics; Animals; Anti-Inflammatory Agents; Disease Models, Animal; Interleukin-1beta; Interleukin-6; Male; Neuralgia; Phenanthrenes; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza; Signal Transduction; Spinal Cord Dorsal Horn; Tumor Necrosis Factor-alpha

There is an urgent need for novel and effective treatment options for acute myeloid leukemia (AML). Triptolide, a diterpenoid tri-epoxide compound isolated from the herb Tripterygium wilfordii and its water-soluble pro-drug-Minnelide have shown promising anti-cancer activity. A recent clinical trial for patients with solid tumors confirmed the safety and efficacy at biologically equivalent doses of 0.2 mg/kg/day and lower.. Cell viability of multiple AML cell lines as well as patient apheresis samples were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) based assay. Apoptosis was evaluated by estimating the amount of cleaved caspase. AML cell line (THP1-Luc) was implanted in immunocompromised mice and treated with indicated doses of Minnelide. Leukemic burden before and after treatment was evaluated by imaging in an In Vivo Imaging System (IVIS).. In the current study, we show that Minnelide, at doses below maximum tolerated dose (MTD) demonstrates leukemic clearance of both primary AML blasts and luciferase expressing THP-1 cells in mice. In vitro, multiple primary AML apheresis samples and AML cell lines (THP-1, KG1, Kasumi-1, HL-60) were sensitive to triptolide mediated cell death and apoptosis in low doses. Treatment with triptolide led to a significant decrease in the colony forming ability of AML cell lines as well as in the expression of stem cell markers. Additionally, it resulted in the cell cycle arrest in the G1/S phase with significant downregulation of c-Myc, a major transcriptional regulator mediating cancer cell growth and stemness.. Our results suggest that Minnelide, with confirmed safety and activity in the clinic, exerts a potent anti-leukemic effect in multiple models of AML at doses easily achievable in patients.

Topics: Animals; Apoptosis; Biomarkers, Tumor; Cell Cycle Checkpoints; Cell Line, Tumor; Disease Models, Animal; Disease Progression; Diterpenes; Down-Regulation; Drug Evaluation, Preclinical; Epoxy Compounds; Humans; Leukemia, Myeloid, Acute; Mice; Neoplastic Stem Cells; Organophosphates; Phenanthrenes; Proto-Oncogene Proteins c-myc; Tumor Burden; Tumor Stem Cell Assay

Cryptotanshinone (CT), a purified compound initially isolated from the dried roots of Salvia militorrhiza. Bunge, exhibits cytotoxic antitumor effects on many tumors. We have shown that CT possesses the dual capacities to concomitantly inhibit the proliferation of lung cancer cells and promote the generation of antitumor immunity. In this study, we investigated whether CT could be used to treat hepatocellular carcinoma (HCC) using a mouse Hepa1-6 model. CT inhibited the proliferation of mouse hepatoma (Hepa1-6) cells in vitro by inducing Hepa1-6 cells apoptosis through the JAK2/STAT3 signaling pathway. In addition, CT activated macrophages and polarized mouse bone marrow-derived macrophages (BMM) toward an M1 phenotype in vitro, which depended on the TLR7/MyD88/NF-κB signaling pathway. Furthermore, CT significantly inhibited the growth of syngeneic Hepa1-6 hepatoma tumors, and, in combination with anti-PD-L1 cured Hepa1-6-bearing mice with the induction of long-term anti-Hepa1-6 specific immunity. Immunoprofiling of treated Hepa1-6-bearing mice revealed that CT-promoted activation of tumor-infiltrating macrophages and dendritic cells, induction of antitumor T cell response, and infiltration of effector/memory CD8 T cells in the tumor tissue. Importantly, the immunotherapeutic effects of CT and anti-PD-L1 depended on the presence of CD8 T cells. Thus, CT and anti-PD-L1 may provide an effective immunotherapeutic regimen for human HCC based on a combination of cytotoxic effects and induction of tumor-specific immunity.

Topics: Adaptive Immunity; Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; B7-H1 Antigen; Carcinoma, Hepatocellular; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Liver Neoplasms; Lymphocyte Activation; Macrophages; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Phenanthrenes; Salvia miltiorrhiza; Signal Transduction; Toll-Like Receptor 7; Treatment Outcome

Cryptotanshinone (CTS) is a natural compound from the Chinese herb Salvia miltiorrhiza. Previous studies demonstrated that CTS possesses anti-apoptotic and anti-inflammatory properties. However, its effects and underlying mechanism on renal ischaemia reperfusion (IR) injury remain unknown. In the present study, we investigated the effects of CTS on renal IR injury and its potential underlying mechanisms. Mice were randomized into four groups as follows: (a) sham operation + vehicle, (b) sham operation + CTS, (c) IR + vehicle, (d) IR + CTS. The CTS-treated group were injected intraperitoneally with CTS (10 mg/kg/d) for 7 days prior to IR operation. Renal IR injury was induced by clamping the bilateral renal artery for 30 minutes followed by 24 hours of reperfusion. The mice were then killed to collect the serum and the kidneys for analysis. The results of the present study showed that CTS pretreatment significantly attenuates IR-induced renal functional and morphological injuries, which was accompanied with inhibition of cell apoptosis and inflammatory response. Moreover, the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and the activation of nuclear factor-κB (NF-κB) signalling were inhibited by CTS. Therefore, CTS could be a useful therapeutic agent in the fight against renal IR injury.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Disease Models, Animal; Humans; Inflammation; Injections, Intraperitoneal; Kidney; Male; Mice; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Phosphorylation; Reperfusion Injury; Signal Transduction; Tumor Necrosis Factor-alpha

Topics: Animals; Atherosclerosis; Diet, High-Fat; Disease Models, Animal; DNA Damage; Humans; Hydrogen Peroxide; Oxidation-Reduction; Oxidative Stress; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Rabbits; Reactive Oxygen Species; Sirtuin 1

Silymarin has been used in the treatment of a number of liver diseases for a long time, but its efficacy in preventing triptolide induced acute hepatotoxicity has not been reported previously. The present study aimed to assess the protective effect of silymarin against triptolide (TP)-induced hepatotoxicity in rats. Rats were orally administrated with silymarin (50, 100 and 200 mg/kg) for 7 days and received intraperitoneal TP (2 mg/kg) on the day 8. Hepatic injuries were comprehensively evaluated in terms of serum parameters, morphological changes, oxidative damage, inflammation and apoptosis. The results demonstrated that TP-induced increases in serum parameters, including alanine transaminase, aspartate aminotransferase, alkaline phosphatase, total cholesterol and γ-glutamyl transpeptidase, which were determined using a biochemical analyzer, and histopathological alterations and hepatocyte apoptosis as determined by hematoxylin and eosin and TUNEL staining, respectively, were prevented by silymarin pretreatment in a dose-dependent manner. TP-induced depletions in the activity of the antioxidant enzymes superoxide dismutase, glutathione peroxidase, glutathione S-transferase and catalase, and glutathione levels, were also significantly reversed by silymarin, as determined using specific kits. Additionally, silymarin dose-dependently exhibited inhibitory effects on malonaldehyde content in the liver. The production of proinflammatory cytokines was investigated using ELISA kits, and the results demonstrated that silymarin dose-dependently inhibited the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10 and IL-1β in the liver. To determine the mechanism of silymarin, western blot analysis was performed to investigate the protein expression of phosphorylated (p)-p38 and p-c-Jun N-terminal kinase (JNK) of the TNF-α induced inflammatory response and apoptotic pathways. Silymarin significantly blocked p38 and JNK phosphorylation and activation. Additionally, the expression of the proapoptotic proteins cytochrome c, cleaved caspase-3 and Bcl-2-associated X was also reduced following treatment with silymarin, as determined by ELISA, western blotting and immunohistochemistry, respectively. In conclusion, silymarin was demonstrated to dose-dependently protect rat liver from TP-induced acute hepatotoxicity, with the high dose (200 mg/kg) achieving a superior effect. This protective effect may be associated with the improvement of antioxidant and

Topics: Animals; Antioxidants; Apoptosis; Apoptosis Regulatory Proteins; Caspase 3; Chemical and Drug Induced Liver Injury; Cytokines; Disease Models, Animal; Diterpenes; Epoxy Compounds; Hepatocytes; Inflammation Mediators; Lipid Peroxidation; Male; Oxidative Stress; Phenanthrenes; Protective Agents; Rats; Reactive Oxygen Species; Silymarin

Idiopathic pulmonary fibrosis (IPF) and tumor are highly similar to abnormal cell proliferation that damages the body. This malignant cell evolution in a stressful environment closely resembles that of epithelial-mesenchymal transition (EMT). As a popular EMT-inducing factor, TGFβ plays an important role in the progression of multiple diseases. However, the drugs that target TGFB1 are limited. In this study, we found that triptolide (TPL), a Chinese medicine extract, exerts an anti-lung fibrosis effect by inhibiting the EMT of lung epithelial cells. In addition, triptolide directly binds to TGFβ and subsequently increase E-cadherin expression and decrease vimentin expression. In in vivo studies, TPL improves the survival state and inhibits lung fibrosis in mice. In summary, this study revealed the potential therapeutic effect of paraquat induced TPL in lung fibrosis by regulating TGFβ-dependent EMT progression.

Topics: Animals; Cell Line; Cell Movement; Disease Models, Animal; Diterpenes; Drugs, Chinese Herbal; Epithelial Cells; Epithelial-Mesenchymal Transition; Epoxy Compounds; Humans; Idiopathic Pulmonary Fibrosis; Lung; Mice; Molecular Docking Simulation; Paraquat; Phenanthrenes; Protein Binding; Transforming Growth Factor beta1

The present study aimed to examine the mechanism of triptolide in the treatment of ankylosing spondylitis (AS) through the anti‑ossification effect of bone morphogenetic protein (BMP)/small mothers against decapentaplegic (Smad) pathway. Male rats were randomly divided into five groups: Normal rat group; model group; triptolide low dose group (10 mg/kg); triptolide middle dose group (20 mg/kg); triptolide high dose group (40 mg/kg). The spinal joint capsules of each group of rats were collected to perform primary cell culture to determine the levels of cell proliferation. Western blot and reverse transcription‑polymerase chain reaction analyses, and ELISA were used to detect the mRNA and protein expression levels of core‑binding factor α1 (Cbfal), BMP receptor type II (BMPRII), Smad1, Smad4, Smad5 and Smad6, the protein expression of phosphorylation indicators, including phosphorylated (p)Smad1 and pSmad5, the mRNA expression of tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑1β and IL‑6 in rat plasma, and the mRNA expression of BMP/Smads in fibroblasts induced by recombinant human (rh)BMP‑2. The effects on AS in the rats were also examined. The results revealed that, following intervention with triptolide, inflammation was suppressed, and the mRNA expression levels of TNF‑α, IL‑1β and IL‑6 were reduced. The expression levels of Cbfal, BMPRII, Smad1, Smad4 and Smad5 were also reduced, whereas the expression of Smad6 was increased. Following induction by rhBMP‑2, the effects of triptolide weakened, with the most marked effects observed at the highest concentration, suggesting that triptolide functions through the BMP/Smad signaling pathway. Taken together, the results suggested that triptolide may be used to treat AS, the mechanism of which may be through the BMP/Smad pathway.

Topics: Animals; Biomarkers; Bone Morphogenetic Proteins; Cell Proliferation; Cytokines; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Fibroblasts; Gene Expression; Immunosuppressive Agents; Male; Osteogenesis; Phenanthrenes; Protein Isoforms; Rats; RNA, Messenger; Signal Transduction; Smad Proteins; Spondylitis, Ankylosing

Topics: Animals; Apoptosis; Cardiotonic Agents; Cells, Cultured; Disease Models, Animal; DNA Methylation; Epigenesis, Genetic; Heart Failure; Humans; Kelch-Like ECH-Associated Protein 1; Mice; Mice, Inbred C57BL; Mice, Knockout; Myocardium; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Promoter Regions, Genetic

Resident fibroblasts that contact tumor epithelial cells (TEC) can become irreversibly activated as cancer-associated-fibroblasts (CAF) that stimulate oncogenic signaling in TEC. In this study, we evaluated the cross-talk between CAF and TEC isolated from tumors generated in a mouse model of KRAS/mut p53-induced pancreatic cancer (KPC mice). Transcriptomic profiling conducted after treatment with the anticancer compound Minnelide revealed deregulation of the TGFβ signaling pathway in CAF, resulting in an apparent reversal of their activated state to a quiescent, nonproliferative state. TEC exposed to media conditioned by drug-treated CAFs exhibited a decrease in oncogenic signaling, as manifested by downregulation of the transcription factor Sp1. This inhibition was rescued by treating TEC with TGFβ. Given promising early clinical studies with Minnelide, our findings suggest that approaches to inactivate CAF and prevent tumor-stroma cross-talk may offer a viable strategy to treat pancreatic cancer.

Topics: Animals; Apoptosis; Cancer-Associated Fibroblasts; Carcinogenesis; Carcinoma, Pancreatic Ductal; Cell Proliferation; Disease Models, Animal; Diterpenes; Epithelial Cells; Epoxy Compounds; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Mice; Mice, Inbred C57BL; Mutation; Organophosphates; Pancreatic Neoplasms; Phenanthrenes; Proto-Oncogene Proteins p21(ras); Signal Transduction; Transforming Growth Factor beta; Tumor Cells, Cultured; Tumor Suppressor Protein p53

The hepatotoxicity of Tripterygium wilfordii Hook. f. (TW), due to the presence of triptolide (TP), limits its therapeutic potential. Based on the traditional Chinese medicine theory, the theory of "Yi lei xiang zhi" was proposed that Chinese herbs with different efficacy can restrict each other to achieve the least adverse reactions.. To observe the effects of Catapol (CAT) and Panax notoginseng saponins (PNS), active ingredients in Rehmannia glutinosa (RG) and Panax notoginseng (PN) respectively, on reducing TP-induced hepatotoxicity, and further to explore the mechanisms.. The human hepatic cell line L-02 was cultured and treated with CAT, PNS or Combinations, and then treated with TP. The cytotoxic assay, the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH), apoptosis, mitochondrial membrane potential and the expressions of NF-E2-related factor 1 (Nrf1) and its downstream targets were detected. Rats were treated with TP, TP + CAT, TP + PNS, or the combinations for 4 weeks. The levels of ALT, AST and LDH in serum, apoptosis of liver cells, mitochondria injury and the protein expressions of Caspase 3 and Nrf1 were investigated.. CAT, PNS or CAT+PNS pre-treatment inhibited TP-induced toxicity in L-02 cells, distinctly decreased the apoptosis, alleviated the reduction of mitochondrial membrane potential, and modulated the expressions of Nrf1 and its downstream target, the mitochondrial transcription factor A (TFAM) and cytochrome C (Cyt-C). CAT, PNS or CAT+PNS inhibited the TP-induced hepatotoxicity in SD rats by reducing the mitochondria injury, decreasing the cells apoptosis and increasing the Nrf1 protein expression. Noticeably, TP + PNS + CAT combinations exhibited more effective than any single ingredient alone.. PNS and CAT were able to effectively attenuate TP-induced hepatotoxicity. The efficiency benefits from their modulating Nrf1 and its downstream genes TFAM and Cyt-C, and further influencing mitochondrial functions and cells apoptosis. The combination is more effective than single ingredient alone.

Topics: Animals; Apoptosis; Biomarkers; Caspase 3; Cell Line; Chemical and Drug Induced Liver Injury; Cytochromes c; Cytoprotection; Disease Models, Animal; Diterpenes; DNA-Binding Proteins; Dose-Response Relationship, Drug; Drug Therapy, Combination; Drugs, Chinese Herbal; Epoxy Compounds; Female; Humans; Liver; Membrane Potential, Mitochondrial; Mitochondria, Liver; Mitochondrial Proteins; NF-E2-Related Factor 1; Panax; Phenanthrenes; Phytotherapy; Plants, Medicinal; Quaternary Ammonium Compounds; Rats, Sprague-Dawley; Saponins; Transcription Factors

The discovery of new therapeutic drugs with the efficacious and safe ability to prevent epidermal hyperplasia is extremely urgent for psoriasis. Cryptotanshinone (CTS), an active component isolated from the root of Salvia miltiorrhiza Bunge, has been reported to have antibacterial and antitumor effects. However, its effects on psoriasis have not been reported. Here, we investigated the therapeutic effects of CTS on imiquimod (IMQ)-induced psoriatic-like skin model and explored the underlying mechanisms. Our results revealed that CTS effectively alleviates IMQ-induced epidermal hyperplasia. In vitro studies also indicated that CTS potently inhibits the growth of keratinocytes. We further found that STAT3, a transcription factor for the cell growth, is the key mediator of CTS on the proliferation of keratinocytes. Taken together, our findings indicated that the curative effects of CTS on psoriasis are accomplished mainly through modulating STAT3, which providing evidences to develop CTS as a potential therapeutic agent for patients with psoriasis.

Topics: Animals; Cell Line; Cell Proliferation; Disease Models, Animal; Drugs, Chinese Herbal; Epidermis; Humans; Hyperplasia; Imiquimod; Keratinocytes; Male; Mice, Inbred C57BL; Phenanthrenes; Psoriasis; STAT3 Transcription Factor

The aim of our current study was to investigate the long-term effect and the mechanism of triptolide in an adult nonorthologous rat model of polycystic kidney disease (PKD). Male wild-type (+/+) and Cy/+ cystic Han:SPRD rats were treated with vehicle or triptolide from 4 to 16 wk of age. Rats were killed at 16 wk of age for blood, urine, and organ collection. Human-derived WT9-12 PKD cells were treated with triptolide with or without IL-6 pretreatment. Cell proliferation, apoptosis, and cytotoxicity were determined. Western blotting and immunohistochemistry analysis were performed to evaluate the activation of IL-6-JAK2-STAT3 pathway. Renal function was protected by 12 wk of triptolide treatment in cystic Han:SPRD rats as shown by reduced blood urea nitrogen, serum creatinine, and proteinuria levels. Cyst and kidney growth were also retarded by triptolide treatment in Cy/+ rats. We further found that the proliferation index was reduced by triptolide in cystic rats, which was correlated with the reduced expression of IL-6/IL-6 receptor, decreased phosphorylation of JAK2-STAT3, and increased expression of suppressor of cytokine signaling 3 (SOCS3). The inhibitory effect of triptolide was further studied in WT9-12 cells. Triptolide inhibited cell proliferation and the activation of JAK2-STAT3 pathway in PKD cells, but it increased the expression of SOCS3. Pretreatment with IL-6 attenuated the inhibitory effect of triptolide on STAT3 phosphorylation. Our study revealed a long-term beneficial effect of triptolide in PKD that was probably through inhibition of the JAK2-STAT3 pathway.

Topics: Animals; Apoptosis; Blood Urea Nitrogen; Cell Line; Cell Proliferation; Creatinine; Disease Models, Animal; Disease Progression; Diterpenes; Epoxy Compounds; Humans; Interleukin-6; Janus Kinase 2; Kidney; Male; Phenanthrenes; Phosphorylation; Polycystic Kidney Diseases; Proteinuria; Rats, Transgenic; Receptors, Interleukin-6; Signal Transduction; STAT3 Transcription Factor; Suppressor of Cytokine Signaling 3 Protein

Triptolide is a structurally unique diterpene triepoxide with potent antitumor activity. However,the effect and mechanism of triptolide on hepatocellular carcinoma (HCC) is not well studied.. Cells were treated with triptolide, and the anti-HCC activity of triptolide was evaluated using flow cytometry, western blot, and xenograft studies. MicroRNA microarray and quantitative reverse-transcription polymerase chain reaction was used to identify differential microRNAs induced by triptolide. Chromatin immunoprecipitation assay was employed to study the interaction between c-Myc and genomic regions of miR106b-25. MicroRNAs overexpression and knockdown experiments were performed to determine the role of these microRNAs in triptolide-induced apoptosis.. Triptolide inhibited cell proliferation and induced marked apoptosis in multiple HCC cell lines with different p53 status. Several signaling molecules involved in different pathways were altered after the treatment of triptolide. Xenograft tumor volume was significantly reduced in triptolide-treated group compared with vehicle control group. Two miRNA clusters, miR-17-92 and miR-106b-25, were significantly suppressed by triptolide, which resulted in the upregulation of their common target genes, including BIM, PTEN, and p21. In HCC samples, high levels of these miRNA clusters correlated with shorter recurrence free survival. Triptolide inhibited the expression of theses miRNAs in a c-Myc-dependent manner, which enhanced triptolide-induced cell death. We further showed that triptolide down-regulated the expression of c-Myc through targeting ERCC3, a newly identified triptolide-binding protein.. The triptolide-induced modulation of c-Myc/miRNA clusters/target genes axis enhances its potent antitumor activity, which indicates that triptolide serves as an attractive chemotherapeutic agent against HCC.

Topics: Animals; Antineoplastic Agents, Alkylating; Carcinoma, Hepatocellular; Cell Death; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Diterpenes; DNA Helicases; DNA-Binding Proteins; Epoxy Compounds; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Mice; MicroRNAs; Oncogenes; Phenanthrenes; Proto-Oncogene Proteins c-myc; RNA Interference; Xenograft Model Antitumor Assays

Intestinal dysfunction, especially acute pathologies linked to intestinal ischemia/reperfusion (I/R) injury, is profoundly affected by inflammation and improper execution of cell death. Few studies have examined the efficacy of combined strategies in regulated intestinal epithelial necrosis after intestinal I/R. Here, we evaluated the functional interaction between poly (adenosine diphosphate-ribose) polymerase 1 (PARP-1)-induced parthanatos and receptor-interacting protein 1/3 (RIP1/3) kinase-induced necroptosis in the pathophysiological course of acute ischemic intestinal injury.. Anesthetized adult male Sprague-Dawley rats were subjected to superior mesenteric artery occlusion consisting of 1.5 h of ischemia and 6 h of reperfusion. The PARP-1-specific inhibitor PJ34 (10 mg/kg) and the RIP1-specific inhibitor Necrostatin-1 (1 mg/kg) were intraperitoneally administered 30 min before the induction of ischemia.. Intestinal I/R was found to result in PARP-1 activation and RIP1/3-mediated necrosome formation. PJ34 or Necrostatin-1 treatment significantly improved the mucosal injury, while the combined inhibition of PARP-1 and RIP1/3 conferred optimal protection of the intestine. Meanwhile, results from terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay showed a decrease in intestinal epithelial cell death. Interestingly, we further showed that PARP-1 might act as a downstream signaling molecule of RIP1 in the process of I/R-induced intestinal injury and that the RIP1/PARP-1-dependent cell death signaling pathway functioned independently of caspase 3 inhibition.. The results of our study provide a molecular basis for combination therapy that targets both pathways of regulated necrosis (parthanatos and necroptosis), to treat acute intestinal I/R-induced intestinal epithelial barrier disruption.

Topics: Animals; Apoptosis; Disease Models, Animal; Drug Therapy, Combination; Epithelial Cells; Humans; Imidazoles; Indoles; Intestinal Mucosa; Male; Necrosis; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Protein Serine-Threonine Kinases; Rats; Rats, Sprague-Dawley; Receptor-Interacting Protein Serine-Threonine Kinases; Reperfusion Injury; Signal Transduction; Treatment Outcome

Metabotropic glutamate receptor (mGlu)

Topics: Animals; Cell Line; Disease Models, Animal; Diterpenes; Dopaminergic Neurons; Epoxy Compounds; Inflammation; Interleukin-1beta; Lipopolysaccharides; Macrophage Activation; Male; Mice; Microglia; Nitric Oxide; Nitric Oxide Synthase Type II; Parkinson Disease; Phenanthrenes; Primary Cell Culture; Rats; Rats, Sprague-Dawley; Receptor, Metabotropic Glutamate 5; Signal Transduction; Transcriptional Activation; Tumor Necrosis Factor-alpha; Up-Regulation

Dihydrotanshinone, tanshinone I, cryptotanshinone, and tanshinone IIA are major lipid-soluble constituents isolated from Salvia miltiorrhiza Bunge (Danshen). In the present study, a systematic method was developed to simultaneously isolate and purify those compounds using macroporous adsorption resins and semi-preparative HPLC with a dynamic axial compress (DAC) system. The Danshen extract (95% alcohol) was divided into three fractions using different concentrations of alcohol (0%, 45%, and 90%) on D101 column. The content of total tanshinones of 90% alcohol eluent (TTS) was over 97%. Furthermore, the anti-inflammatory effects of those samples were investigated on LPS-stimulated RAW264.7 cells and three animal models. The results showed that the anti-inflammatory effect of TTS in vitro was superior to the one of any other sample including 0% and 45% eluent, and total tanshinones capsules. In addition, TTS exhibited a stronger anti-inflammatory effect than that of dihydrotanshinone, tanshinone IIA, cryptotanshinone, and tanshinone I, respectively. For animal models, TTS could significantly suppress xylene-induced ear oedema and rescue LPS-induced septic death and acute kidney injury in mice. In summary, the separation process developed in the study was high-efficiency, economic, and low-contamination, which was fit to industrial producing. TTS is a potential agent for the treatment of inflammatory diseases.

Topics: Abietanes; Acute Kidney Injury; Animals; Anti-Inflammatory Agents; Chromatography, High Pressure Liquid; Cytokines; Disease Models, Animal; Kidney; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred BALB C; Nitric Oxide; Phenanthrenes; RAW 264.7 Cells; Salvia miltiorrhiza; Sepsis

To discuss the curative effect and security of triptolide (TPL) on TNF transgenic (TNF-Tg) mice with rheumatoid arthritis (RA), and to explore the mechanism primarily.. 40 TNF-Tg RA mice were randomlydivided into five groups averagely: the control group, low-dose group (3.3 μg/kg/d TPL), middle-dose group (10 μg/kg/d TPL), high-dose group (33 μg/kg/d TPL) and MTX group (0.1 mg/kg/d MTX). Mice were administrated five days a week for six weeks. The arthritis deformation index, arthritis detumescencepercentage and the level of inflammatory factor in each group were recorded during theadministration. After administration, body weight, liver and renal function indexes, the apoptosis rates of osteoclast precursors (OCP), T and B lymphocytes in the peripheral blood and the number of osteoclast (OC) were detected and compared. μCT scanning and HE staining methods were taken to observethebone histomorphometry and bony erosion.. After administration, the arthritis deformation indexes were lower and arthritis detumescence percentageswere higher in TPL groups thanthe control group (P < 0.05), and the arthritis detumescence percentage in the high-dose group was higher than the MTX group (P < 0.05). The liver function index ALT increased after administrationin the high-dose group but was lower than that in the MTX group (P < 0.05). The level of IL-1α, IL-1β, and TNF-α decreased in the TPL groups and MTX group after administration;The apoptosis rates of OCP and T lymphocytes in middle and high dose TPL groups and MTX group were higher than other groups, and that in the high-dose group was higher than the MTX group (P < 0.05). Compared with the other groups, the bony erosion degree was lower and the number of OC was less and the parameters of bone histomorphometry were better in the high-dose group.. TPL could improvearthritic of TNF-Tg mice by decreasing the levels of pro-inflammatory cytokines, promoting the apoptosis of OCP, inhibiting the generation of OC and bone resorption. There was some toxic and side effect on liver for high-dose TPL which was weaker than the MTX.

Topics: Animals; Antirheumatic Agents; Apoptosis; Arthritis, Rheumatoid; Biomarkers; Bone Resorption; Chemical and Drug Induced Liver Injury; Cytokines; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Inflammation Mediators; Joints; Lymphocytes; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Transgenic; Osteoclasts; Phenanthrenes; Risk Assessment; Time Factors; Tumor Necrosis Factor-alpha

Ovarian cancer is the most malignant gynecologic cancer among women worldwide. Cryptotanshinone (CT), isolated from Salvia miltiorrhiza Bunge, has been identified as a potential therapeutic agent in treating several malignant tumors, but the molecular mechanism of CT in ovarian cancer still remains illustrated. Here, we sought to elucidate the regulatory function of CT on cell glucose metabolism in ovarian cancer. The treatment of CT on ovarian cancer cells effectively inhibited glucose uptake and lactate production in ovarian cancer cells. The expression levels of glycolysis-related proteins, such as GLUT1, LDHA, and HK2, were decreased by the treatment of CT detected by qRT-PCR and immunoblotting. Mechanistically, CT exerted its anti-tumor effect by targeting STAT3/SIRT3/HIF-1α signaling pathway in vitro and in vivo, which could be rescued by the introduction of SIRT3 shRNA in ovarian cancer cells. The clinical data showed that the expression level of STAT3 in ovarian cancer patients' sera and tissues was positively correlated with those of GLUT1, LDHA, HK2 and HIF-1α, but negatively with that of SIRT3These findings provide evidence that CT inhibited cellular glycolysis-induced cell growth and proliferation through repression of STAT3/SIRT3/HIF-1α signaling pathway, indicating that CT may be developed as a chemotherapeutic agent to treat ovarian cancer.

Topics: Animals; Antineoplastic Agents, Phytogenic; Biomarkers; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Drugs, Chinese Herbal; Energy Metabolism; Female; Genes, Reporter; Glucose; Glycolysis; Humans; Mice; Ovarian Neoplasms; Phenanthrenes; Promoter Regions, Genetic; Signal Transduction; Sirtuin 3; STAT3 Transcription Factor; Xenograft Model Antitumor Assays

Higher levels of hyaluronan (HA) and its receptors CD44 and RHAMM have been associated with poor prognosis and metastasis in NSCLC. In the current study, our goal was to define, using cellular and orthotopic lung tumor models, the role of HA-CD44/RHAMM signaling in lung carcinogenesis and to assess the potential of triptolide to block HA-CD44/RHAMM signaling and thereby suppress the development and progression of lung cancer. Triptolide reduced the viability of five non-small cell lung cancer (NSCLC) cells, the proliferation and self-renewal of pulmospheres, and levels of HA synthase 2 (HAS2), HAS3, HA, CD44, RHAMM, EGFR, Akt and ERK, but increased the cleavage of caspase 3 and PARP. Silencing of HAS2, CD44 or RHAMM induced similar effects. Addition of excess HA to the culture media completely abrogated the effects of triptolide and siRNAs targeting HAS2, CD44, or RHAMM. In an orthotopic lung cancer model in nude rats, intranasal administration of liposomal triptolide (400 μg/kg) for 8 weeks significantly reduced lung tumor growth as determined by bioluminescence imaging, lung weight measurements and gross and histopathological analysis of tumor burden. Also, triptolide suppressed expressions of Ki-67, a marker for cell proliferation, HAS2, HAS3, HA, CD44, and RHAMM in lung tumors. Overall, our results provide a strong rationale for mitigating lung cancer by targeting the HA-CD44/RHAMM signaling axis.

Topics: Animals; Antineoplastic Agents, Alkylating; Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Disease Models, Animal; Diterpenes; Epoxy Compounds; Extracellular Matrix Proteins; Gene Silencing; Humans; Hyaluronan Receptors; Hyaluronan Synthases; Lung Neoplasms; Male; Phenanthrenes; Rats; RNA, Small Interfering; Signal Transduction; Xenograft Model Antitumor Assays

The aim of the present study was to investigate the effects of sodium tanshinone IIA sulfonate (STS) on inflammatory responses, aortic endothelial cell apoptosis, disseminated intravascular coagulation (DIC) and multiple organ damage in an animal model of classic heat stroke (CHS). The rats in the heat stroke (HS) and STS‑treated heat stroke (STS‑HS) groups were placed into a pre‑warmed animal temperature controller (ATC) at 35˚C. The moment at which the rectal temperature reached 43.5˚C was considered as the time of onset of HS. In the HS groups, the rats were removed from the ATC and allowed to recover at 26˚C for 0, 2, 6 or 12 h. In the STS‑HS groups, the rats received femoral vein injections of 5‑40 mg/kg STS immediately following the onset of HS and were subsequently placed at a temperature of 26˚C to recover for 6 h. In the present study, the serum levels of tumor necrosis factor (TNF)‑α, interleukin (IL)‑1β and IL‑6 were assessed using ELISA, and the numbers of apoptotic aortic endothelial cells were investigated using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick‑end labeling combined with immunofluorescence. In the HS groups, the serum levels of TNF‑α, IL‑1β and IL‑6, as well as the numbers of apoptotic aortic endothelial cells were increased compared with the normothermic control group. Additionally, the plasma prothrombin time, activated partial thromboplastin time and D‑dimer level were significantly increased in the HS group compared with the normothermic control group following recovery for 6 h. By contrast, the platelet count was decreased in the HS group compared with the normothermic control group. The serum levels of creatinine, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were increased and histopathological damage to multiple organs was observed in the HS group following recovery for 6 h. In the STS‑HS groups, cytokine levels and apoptotic aortic endothelial cell numbers were reduced compared with the HS group after 6 h recovery. STS (40 mg/kg) treatment additionally improved the serum levels of organ injury indicators and plasma indicators of coagulopathy, and prevented histopathological damage to multiple organs. These findings demonstrated that STS treatment may ameliorate multiple organ damage by attenuating inflammatory responses, aortic endothelial cell apoptosis and DIC in CHS. These results suggested that STS may hold potential a

Topics: Animals; Apoptosis; Biomarkers; Cytokines; Disease Models, Animal; Disseminated Intravascular Coagulation; Endothelial Cells; Heat Stroke; Inflammation; Male; Phenanthrenes; Rats

Poly(ADP-ribose) polymerase (PARP), calpain, and nuclear factor-κB (NF-κB) are reported to participate in inflammatory reactions in pathologic conditions and are involved in traumatic brain injury. The objective of this study was to investigate whether PARP participates in inflammation related to calpain and NF-κB in a mouse model of controlled cortical impact (CCI).. PJ34 (10 mg/kg), a selective PARP inhibitor, was administered intraperitoneally 5 minutes and 8 hours after experimental CCI. We then performed a histopathologic analysis, and we measured calpain activity and protein levels in all animals. The cytosolic, mitochondria, and nuclear fractions were prepared and used to determine the levels of PARP, calpastatin, NF-κB p65, inhibitory-κB-α, tumor necrosis factor-α, interleukin-1β, intracellular adhesion molecule-1, inducible nitric oxide synthase, and cyclooxygenase-2. We then measured blood-brain barrier disruption using electron microscopy at 6 and 24 hours after CCI.. Treatment with PJ34 markedly reduced the extent of both cerebral contusion and edema, improved neurologic scores, and attenuated blood-brain barrier damage resulting from CCI. Our data showed that the cytosolic and nuclear fractions of calpain and NF-κB were up-regulated in the injured cortex and that these changes were reversed by PJ34. Moreover, PJ34 significantly enhanced the calpastatin and inhibitory-κB levels and decreased the levels of inflammatory mediators.. PARP inhibition by PJ34 suppresses the overactivation of calpain and the production of inflammatory factors that are caused by NF-κB activation and attenuates neuronal cell death in a mouse model of CCI.

Topics: Animals; Brain Injuries, Traumatic; Calpain; Disease Models, Animal; Down-Regulation; Inflammation Mediators; Male; Mice; Mice, Inbred BALB C; Neuroprotective Agents; NF-kappa B; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Random Allocation

It has been established that signal transducer and activator of transcription 3 serves as an oncoprotein in various human cancers; targeting it is therefore a reasonable approach for emerging cancer therapies. Cryptotanshinone, a natural compound extracted from the root of Salvia miltiorrhiza Bunge, has been identified as a potential STAT3 inhibitor. However, its functional role in renal cell carcinomas remains largely unknown. Therefore, we investigated the mode of action for cryptotanshinone. We found that cryptotanshinone substantially suppressed cancer cell growth while it promoted cell apoptosis by inhibiting the phosphorylation of STAT3 at Tyr705 and its blocking nuclear translocation. Coordinately, P-AKT, CyclinD1, C-MYC, MEKK2, and HGF were down-regulated and cell cycle progression was arrested at the G0/G1 phase, thereby attenuating cell proliferation. Moreover, the level of Cleaved-Caspase-3 was elevated while Bcl-2 and Survivin were down-regulated, accounting for the increased apoptosis. Furthermore, in vivo results revealed that cryptotanshinone effectively inhibits tumorigenesis in an A498-xenografted mouse model. Taken together, our data gives a more comprehensive understanding of how cryptotanshinone functions in renal cell carcinomas and demonstrates its potential as a powerful therapeutic approach to treat renal cell carcinomas.

Topics: Animals; Apoptosis; Carcinoma, Renal Cell; Caspase 3; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Drugs, Chinese Herbal; Genes, myc; Humans; Kidney Neoplasms; Male; Mice; Models, Biological; Phenanthrenes; Phosphorylation; Protein Transport; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; STAT3 Transcription Factor; Xenograft Model Antitumor Assays

Osteoarthritis (OA) is a common degenerative disease characterized by progressive erosion of articular cartilage, subchondral bone sclerosis and synovitis. Cryptotanshinone (CTS), an active component extracted from the root of Salvia miltiorrhiza Bunge, has been shown to have potent anti-inflammatory effects. However, its effects on OA have not been clearly elucidated. This study aimed to assess the effect of CTS on human OA chondrocytes and mice OA models. Human OA chondrocytes were pretreated with CTS (5, 10 and 20μM) for 2h and subsequently stimulated with IL-1β for 24h. Production of NO, PGE2, IL-6, TNF-α was evaluated by the Griess reaction and ELISA. The protein expression of COX-2, iNOs, MMP-3, MMP13, COX-2, ADAMTS-5, JNK, p-JNK, ERK, p-ERK, p38, p-p38, p-IKKα/β, p65, p-p65, IκB-α, and p-IκB-α was tested by Western blot. In vivo, the severity of OA was determined by histological analysis. We found that CTS significantly inhibited the IL-1β-induced production of NO and PGE2; expression of COX-2, iNOS, MMP-3, MMP-13, and ADAMTS-5. Furthermore, CTS in dramatically suppressed IL-1β-stimulated NF-κB and MAPK activation. Immunofluorescence staining demonstrated that CTS could suppress IL-1β-induced phosphorylation of p65 nuclear translocation. In vivo, treatment of CTS prevented the destruction of cartilage and the thickening of subchondral bone in mice OA models. These results indicate that the therapeutic effect of CTS on OA is accomplished through the inhibition of both NF-κB and MAPK signaling pathways. Our findings provide the evidence to develop CTS as a potential therapeutic agent f or patients with OA.

Topics: Animals; Anti-Inflammatory Agents; Cells, Cultured; Chondrocytes; Cyclooxygenase 2; Dinoprostone; Disease Models, Animal; Disease Progression; Female; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Middle Aged; NF-kappa B; Nitric Oxide; Osteoarthritis; Phenanthrenes; Salvia miltiorrhiza; Signal Transduction; Tumor Necrosis Factor-alpha

Optic nerve crush model could be used to investigate the mechanism of neuronal survival and axonal regeneration in central nervous system. Triptolide, a Chinese herb extract with anti-inflammatory and immunosuppressive activities, has shown neuron protective functions in nervous system. In this study, we investigated the changes in retinal ganglion cell survival and axonal regeneration after administration of triptolide in optic nerve crush model. Triptolide treatment tended to promote retinal ganglion cell survival rather than optic nerve regeneration as well as inhibit the expression of tumor necrosis factor-α and activation of nuclear factor-kappa B. These findings suggested that intraperitoneal injection of triptolide may be an effective treatment for optic nerve injury and this effect was attributed at least in part to its anti-inflammatory actions.

Topics: Animals; Axons; Cell Nucleus; Cell Survival; Disease Models, Animal; Diterpenes; Epoxy Compounds; Mice, Inbred C57BL; Nerve Crush; Nerve Regeneration; Optic Nerve Injuries; Phenanthrenes; Protein Transport; Retinal Ganglion Cells; Transcription Factor RelA; Tumor Necrosis Factor-alpha

The weakened tumour colonization of attenuated

Topics: Animals; Biological Therapy; Combined Modality Therapy; Disease Models, Animal; Diterpenes; Epoxy Compounds; Immunosuppressive Agents; Melanoma; Mice, Inbred C57BL; Necrosis; Neovascularization, Pathologic; Neutrophils; Phenanthrenes; Salmonella; Treatment Outcome; Vascular Endothelial Growth Factor A

Triptolide is a vine extract used in traditional Chinese medicines and associated with hepatotoxicity. In vitro data suggest that inhibition of RNA synthesis may be the mechanism of toxicity. For studying drug-induced liver injury the zebrafish has experimental, practical and financial advantages compared with rodents. The aim of this study was to explore the mechanism of triptolide toxicity using zebrafish as the model system. The effect of triptolide exposure on zebrafish larvae was determined with regard to mortality, histology, expression of liver specific microRNA-122 and liver volume. Fluorescent microscopy was used to track toxicity in the Tg(-2.8lfabp:GFP)as3 zebrafish line. Informed by microscopy, RNA-sequencing was used to explore the mechanism of toxicity. Triptolide exposure resulted in dose-dependent mortality, a reduction in the number of copies of microRNA-122 per larva, hepatocyte vacuolation, disarray and oncotic necrosis, and a reduction in liver volume. These findings were consistent across replicate experiments. Time-lapse imaging indicated the onset of injury was 6 h after the start of exposure, at which point, RNA-sequencing revealed that 88% of genes were down-regulated. Immune response associated genes were up-regulated in the triptolide-treated larvae including nitric oxide synthase. Inhibition of nitric oxide synthase increased mortality. Triptolide induces hepatotoxicity in zebrafish larvae. This represents a new model of drug-induced liver injury that complements rodents. RNA sequencing, guided by time-lapse microscopy, revealed early down-regulation of genes consistent with previous invitro studies, and facilitated the discovery of mechanistic inflammatory pathways.

Topics: Animals; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Diterpenes; Epoxy Compounds; Larva; Liver; MicroRNAs; Microscopy, Fluorescence; Phenanthrenes; Polymerase Chain Reaction; Sequence Analysis, RNA; Transcriptome; Zebrafish

Considerable attention has been paid to the introduction of novel naturally occurring plant-derived radiosensitizer compounds in order to augment the radiation efficacy and improve the treatment outcome of different tumors. This study was therefore undertaken to evaluate the antitumor, antiangiogeneic, and synergistic radiosensitizing effects of apigenin, a dietary flavonoid, and/or cryptotanshinone, a terpenoid isolated from the roots of Salvia miltiorrhiza, against the growth of solid Ehrlich carcinoma in female mice. Apigenin (50 mg/kg body weight) and/or cryptotanshinone (40 mg/kg body weight) was intraperitoneally (i.p.) injected into non-irradiated or γ-irradiated (6.5 Gy whole-body γ-irradiation) solid Ehrlich carcinoma-bearing mice for 30 consecutive days. Investigations included molecular targets involved in proliferation, inflammation, angiogenesis, and tumor invasiveness. Treatment with apigenin and/or cryptotanshinone significantly suppressed the growth of solid Ehrlich carcinoma tumors and demonstrated a synergistic radiosensitizing efficacy together with γ-irradiation. These effects were achieved through downregulating the expression of angiogenic and lymphangiogenic regulators, including signal transducer and activator of transcription 3, vascular endothelial growth factor C, and tumor necrosis factor alpha, suppressing matrix metalloproteinase-2 and -9 activities, which play a key role in tumor invasion and metastasis, and enhancing apoptosis via inducing cleaved caspase-3 and granzyme B levels. Histological findings of solid Ehrlich carcinoma tumors verified the recorded data. In conclusion, a synergistic radiosensitizing efficacy for apigenin and cryptotanshinone was demonstrated against Ehrlich carcinoma in the current in vivo murine model, representing therefore a potential therapeutic strategy for increasing the radiation response of solid tumors.

Topics: Animals; Apigenin; Apoptosis; Carcinoma, Ehrlich Tumor; Cell Proliferation; Disease Models, Animal; Female; Gamma Rays; Humans; Mice; Phenanthrenes; Radiation-Sensitizing Agents; Whole-Body Irradiation

Cerebral stroke is a kind of acute cerebrovascular disease with high incidence, morbidity and disability. Treatments against various types of cerebral stroke are limited at preventive measurements due to the lack of effective therapeutic method. The present study aimed to investigate the protective effect of cryptotanshinone (CPT) on cerebral stroke, and investigate the possible mechanism involved in order to develop a novel therapy against stoke. The phosphoinositide 3‑kinase membrane translocation of cerebral stroke rats pretreated with CPT at various concentrations were measured, as well as the phosphorylation of protein kinase B (AKT) and endothelial nitric oxide synthase (eNOS). Additionally, the expression level of B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax) and vascular endothelial growth factor were also assessed using western blotting and reverse transcription‑quantitative polymerase chain reaction. Furthermore, biochemical tests were used to measure the activity of superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO) in both the cerebral cortex and peripheral blood. As a result, CPT‑pretreated rats presented declined phosphoinositide 3‑kinase (PI3K) and AKT expression levels, indicating that the PI3K/AKT signaling pathway was inhibited. Increased Bcl‑2 and NO levels in both the cerebral cortex and peripheral blood demonstrated the anti‑apoptosis and blood vessel protection effect of CPT. Furthermore, increased SOD activity and declined MDA levels demonstrated suppressed lipid peroxidation. In conclusion, CPT exhibited a protective effect against cerebral stroke through inhibition of the PI3K/AKT‑eNOS signaling pathway. These results suggested the potential of CPT as a promising agent in the treatment of cerebral stroke.

Topics: Animals; bcl-2-Associated X Protein; Disease Models, Animal; Humans; Nitric Oxide; Nitric Oxide Synthase Type III; Oncogene Protein v-akt; Phenanthrenes; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-bcl-2; Rats; Signal Transduction; Stroke

Mitochondrial dysfunction, oxidative stress, inflammation, and metabolic reprograming are crucial contributors to hepatic injury and subsequent liver fibrosis. Poly(ADP-ribose) polymerases (PARP) and their interactions with sirtuins play an important role in regulating intermediary metabolism in this process. However, there is little research into whether PARP inhibition affects alcoholic and non-alcoholic steatohepatitis (ASH/NASH).. We investigated the effects of genetic deletion of PARP1 and pharmacological inhibition of PARP in models of early alcoholic steatohepatitis, as well as on Kupffer cell activation in vitro using biochemical assays, real-time PCR, and histological analyses. The effects of PARP inhibition were also evaluated in high fat or methionine and choline deficient diet-induced steatohepatitis models in mice.. Our results suggests that PARP inhibition is a promising therapeutic strategy in steatohepatitis with high translational potential, considering the availability of PARP inhibitors for clinical treatment of cancer.. Poly(ADP-ribose) polymerases (PARP) are the most abundant nuclear enzymes. The PARP inhibitor olaparib (Lynparza) is a recently FDA-approved therapy for cancer. This study shows that PARP is overactivated in livers of subjects with alcoholic liver disease and that pharmacological inhibition of this enzyme with 3 different PARP inhibitors, including olaparib, attenuates high fat or alcohol induced liver injury, abnormal metabolic alteration, fat accumulation, inflammation and/or fibrosis in preclinical models of liver disease. These results suggest that PARP inhibition is a promising therapeutic strategy in the treatment of alcoholic and non-alcoholic liver diseases.

Topics: Animals; Diet, High-Fat; Disease Models, Animal; Fatty Acids; Fatty Liver, Alcoholic; Humans; Kupffer Cells; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; NAD; Nitrosative Stress; Non-alcoholic Fatty Liver Disease; Oxidative Stress; Phenanthrenes; Phthalazines; Piperazines; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Quinolines; Sirtuin 1

The aim of this study was to investigate the therapeutic effects of Tripterygium wilfordii polyglycosidium (TWP) to rats with dextran sulfate sodium (DSS)-induced pouchitis and its possible mechanism.. Sprague-Dawley rats underwent surgery of ileal pouch anal anastomosis (IPAA) and pouchitis was induced by DSS. Rats were randomly divided into no intervention (NI), normal saline (NS) and TWP groups. Rats were lavaged with normal saline (3ml/day in NS group) or TWP (12mg/kg/day in TWP group) for 7days. General conditions of animals and histopathological examinations were evaluated. Interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor (TNF)-α mRNA expression was measured. Levels of occludin and Zo-1 proteins were measured by immunohistochemistry. In addition, ALT and AST were assessed.. TWP significantly attenuated the symptoms of pouchitis characterized by body weight loss, diarrhea, and bloody stool. Furthermore, TWP diminished histological damage compared with other groups. There was a significant reduction in levels of IL-1β, IL-6, and TNF-α, as well as an increase in IL-10 in the TWP group. The expression of tight junction proteins occludin and Zo-1 were increased in the TWP group. There were no statistical differences in serum ALT and AST among the three groups.. TWP significantly ameliorated pouchitis and inhibited the production of IL-1β, IL-6, and TNF-α as well as increased the levels of IL-10, occludin, and Zo-1 protein in rats. These findings suggest TWP might be a potential therapeutic agent for patients with pouchitis.

Topics: Animals; Anti-Inflammatory Agents; Cytokines; Dextran Sulfate; Disease Models, Animal; Diterpenes; Epoxy Compounds; Humans; Ileum; Intestinal Mucosa; Male; Occludin; Phenanthrenes; Pouchitis; Rats; Rats, Sprague-Dawley; Tripterygium; Zonula Occludens-1 Protein

The traditional Chinese medicine (Tripterygium wilfordiiHook.f., TWH) has been clinically used to treat primary and secondary renal diseases and proteinuria for nearly 40 years. However, there is a rare literature about the effect of triptolide (the main active ingredient of TWH) on the expression of oxidative carbonyl protein (OCP) in diabetic nephropathy (DN). This study aimed to provide experimental evidence for triptolide treatment on DN through its effect on the expression of OCP, in order to investigate the effects of triptolide on the expression of OCP in rats with DN. Sixty SD rats were randomly divided into five groups: control group, high-dose triptolide (Th) group, low-dose triptolide (Tl) group, DN model group, and positive control (benazepril) group. The DN model was established using streptozotocin. Urinary protein excretion, fasting blood glucose (FBG), superoxide dismutase (SOD) in renal homogenate, malondialdehyde (MDA) in renal homogenate and renal nitrotyrosine by immunohistochemistry, and the expression of OCP by oxyblotimmune blotting were detected. In the DN model group, rat urinary protein excretion and renal MDA were significantly increased, while renal SOD significantly decreased and nitrotyrosine expression was obviously upregulated in the kidney. After triptolide treatment, 24-h urinary protein excretion (61.96±19.00 vs. 18.32±4.78 mg/day, P<0.001), renal MDA (8.09±0.79 vs. 5.45±0.68 nmol/L, P<0.001), and nitrotyrosine expression were decreased. Furthermore, renal OCP significantly decreased, while renal SOD (82.50±19.10 vs. 124.00±20.52 U/L, P<0.001) was elevated. This study revealed that triptolide can down-regulate the expression of OCP in the renal cortex of DN rats.

Topics: Animals; Blood Glucose; Diabetic Nephropathies; Disease Models, Animal; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Gene Expression Regulation; Humans; Kidney Cortex; Oxidative Stress; Phenanthrenes; Protein Carbonylation; Proteins; Random Allocation; Rats; Rats, Sprague-Dawley; Streptozocin; Superoxide Dismutase

Chronic pain and depression frequently coexist in clinical setting, and current clinical treatments for this comorbidity have shown limited efficacy. Triptolide (T10), an active component of Tripterygium wilfordii Hook F., has been demonstrated to exert strong analgesic activities in experimental pain models, but whether it possesses anti-depressive actions remains unknown. Using a depression comorbidity of chronic pain rat model induced by spinal nerve ligation (SNL), we investigated the potency of T10 for the treatment of comorbid depression in comparison with a widely used antidepressant, fluoxetine (FLX). Concomitant neuroinflammation changes were also examined in the hippocampus. The results showed that prophylactic and reversal treatments with T10 dose-dependently (30, 100, 300μg/kg) inhibited the depression-like behaviors (DLB) assessed by the forced swim test, sucrose preference test and body weight measurement. The anti-depressive efficacy of T10 at 300μg/kg was significantly stronger than that of FLX at 18mg/kg. T10 at all three doses exhibited more efficient analgesic effects than FLX at 18mg/kg. The combined application of T10 with FLX markedly augmented the effects of T10 or FLX per se, with the facilitating effects of T10 at 30μg/kg being most prominent. In addition, nerve injury caused the activation of microglia and p38 MAPK, the upregulation of IL-1β and TNF-α as well as the downregulation of IL-10 in the hippocampus at postoperative week (POW) 3. These neuroinflammatory responses were reversed by subchronic treatment with T10. Taken together, these results demonstrate that T10 possesses potent anti-depressive function, which is correlated with its immunoregulation in the hippocampus. The combination of a low dose of T10 with FLX may become a more effective medication strategy for the treatment of comorbid depression and chronic pain.

Topics: Analgesics; Animals; Antidepressive Agents; Behavior, Animal; Chronic Pain; Depression; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Drug Therapy, Combination; Encephalitis; Epoxy Compounds; Fluoxetine; Hippocampus; Hyperalgesia; Inflammation Mediators; Male; Microglia; Phenanthrenes; Rats, Sprague-Dawley

The serious side effect of Adriamycin (ADR) is cardiomyopathy. Cryptotanshinone (CRY) is widely and safely used as antioxidant with MTD more than 5 mg/g in rats (p.o).. The objective of this study is to study the protection effects of CRY against ADR-induced mitochondrial dysfunction in cardiomyocytes.. The chemical administration lasted for 20 days with an effective dose of CRY (p.o.) at 50 mg/kg in rats. Mitochondrial respiratory chain complex activities, ATP generation, mitochondrial membrane potential (MMP), superoxide anion free radical, oxidative stress-relative enzymes, and mitochondrial biogenesis-relative factors in normal control, ADR (i.p., 1.25 mg/kg), and ADR (i.p., 1.25 mg/kg) + CYP (p.o., 50 mg/kg) groups were detected.. 50 mg/kg CRY significantly promoted the energy production of ATP (16.99 ± 2.38 nmol/g Pro) (Pro: Protein) by increasing the complexes activities except II (p > 0.05). After the treatment of CRY, the suppressed MMP was increased while superoxide anion free radical (0.57 ± 0.07/mg Pro) was inhibited markedly. Mitochondrial biogenesis-relative factors PGC-1α, NRF-1, and TFAM were also promoted. Remarkable augmentations of NO, inducible nitric oxide synthase (iNOS), and increased activity of GSH-PX (p < 0.05) were also detected after the treatment of CRY, while no obvious changes on the activity of nitric oxide synthase (cNOS; p > 0.05) were observed.. These results suggest that CRY protects against ADR-induced mitochondrial dysfunction in cardiomyocytes. It could be an ideal potential drug of cardioprotection.

Topics: Adenosine Triphosphate; Animals; Cardiomyopathies; Cardiotoxicity; Disease Models, Animal; Doxorubicin; Drugs, Chinese Herbal; Male; Membrane Potential, Mitochondrial; Mitochondria, Heart; Myocytes, Cardiac; Oxidative Stress; Phenanthrenes; Rats, Wistar; Salvia miltiorrhiza

Triptolide is a bioactive component of Chinese herbal plant Tripterygium wilfordii Hook F that has recently been noted to attenuate hepatic and cerebral ischemia/reperfusion (I/R) injuries in rodents. To investigate whether triptolide could protect against myocardial I/R injuries, triptolide (25, 50 or 100 μg/kg) was administrated in Wistar rats that underwent left anterior descending coronary artery ligation in this study. Our data showed that triptolide pretreatment could attenuate myocardial infarction, increase the fractional shortening and left ventricular systolic pressure and decrease the left ventricular end-diastolic pressure in ischemic rats. Also, triptolide was noted to inhibit the activities of lactate dehydrogenase and creatine kinase in I/R rats. Moreover, triptolide administration suppressed macrophage infiltration, inhibited the overproduction of tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and malondialdehyde (MDA) in reperfused myocardium tissues and upregulated the activities of antioxidative superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GPx). In addition, nuclear accumulation of nuclear factor erythroid 2-related factor 2 (Nrf2) and the activity of its downstream target Heme oxygenase-1 (HO-1) in ischemic myocardium tissues were enhanced by triptolide pretreatment. In addition, the HO-1 inhibitor, zinc protoporphyrin-IX, abrograted the cardiac protection mediated by triptolide. Our study reveals a novel cardioprotective effect of triptolide in rats with I/R injuries, wherein the activation of Nrf2/HO-1 signaling was involved.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Biomarkers; Cardiotonic Agents; Cytokines; Cytoprotection; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Enzyme Inhibitors; Epoxy Compounds; Heme Oxygenase (Decyclizing); Inflammation Mediators; Macrophages; Myocardial Contraction; Myocardial Infarction; Myocardial Reperfusion Injury; Myocardium; NF-E2-Related Factor 2; Oxidative Stress; Phenanthrenes; Rats, Wistar; Signal Transduction; Ventricular Function, Left

Stroke is the leading cause of neurological disability in humans. Middle cerebral artery occlusion (MCAO) followed by reperfusion is widely accepted to mimic stroke in basic medical research. Triptolide is one of the major active components of the traditional Chinese herb Tripterygium wilfordii Hook F, and has been reported to have potent anti-inflammatory and immunosuppressive properties. Since its preclinical effects on stroke were still unclear, we decided to study the effects of Triptolide on focal cerebral ischemia/reperfusion injury in this study. The results showed that Triptolide treatment significantly attenuates brain infarction volume, water content, neurological deficits, and neuronal cell death rate, which were increased in the MCAO model rats. Immunohistochemistry was used to analyze the expression of glial fibrillary acidic protein (GFAP), Cyclooxygenase-2 (COX-2), inducible nitric oxide (iNOS), and NF-κB in the ischemic brains. The administration of Triptolide showed down-regulation of the iNOS, COX-2, GFAP, and NF-κB expression in MCAO rats. It also increased the expression of bcl-2, and suppressed levels of bax and caspase-3 compared with the MCAO group. Our findings revealed that Triptolide exerts its neuroprotective effects against inflammation with the involvement of inhibition of NF-κB activation.

Topics: Animals; Anti-Inflammatory Agents; Antineoplastic Agents, Alkylating; Apoptosis; Blotting, Western; Cyclooxygenase 2; Disease Models, Animal; Diterpenes; Epoxy Compounds; Glial Fibrillary Acidic Protein; Immunoenzyme Techniques; In Situ Nick-End Labeling; Infarction, Middle Cerebral Artery; Male; Neuroprotective Agents; NF-kappa B; Nitric Oxide; Phenanthrenes; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; Reperfusion Injury; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Severity of Illness Index; Signal Transduction

Prostaglandin E2 (PGE2), one of the terminal products in the cyclooxygenase pathway, plays an important role in various inflammatory responses. To determine whether selective inhibition of PGE2 may relieve these inflammatory symptoms, we synthesized a selective PGE2 synthesis inhibitor, compound A [1-(6-fluoro-5,7-dimethyl-1,3-benzothiazol-2-yl)-N-[(1S,2R)-2-(hydroxymethyl)cyclohexyl]piperidine-4-carboxamide], then investigated the effects on pyrexia, arthritis and inflammatory pain in guinea pigs. In LPS-stimulated guinea pig macrophages, compound A selectively inhibited inducible PGE2 biosynthesis in a dose-dependent manner whereas enhanced the formation of thromboxane B2 (TXB2). Compound A suppressed yeast-evoked PGE2 production selectively and enhanced the production of TXB2 and 6-keto PGF1αin vivo. In addition, compound A relieved yeast-induced pyrexia and also suppressed paw swelling in an adjuvant-induced arthritis model. The effect on gastrointestinal (GI) ulcer formation was also evaluated and compound A showed a lower GI adverse effect than indomethacin. However, compound A failed to relieve yeast-induced thermal hyperalgesia. These results suggest that selective inhibition of PGE2 synthesis may have anti-pyretic and anti-inflammatory properties without GI side effect, but lack the analgesic efficacy.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Benzothiazoles; Depression, Chemical; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Fever; Guinea Pigs; Imidazoles; Indomethacin; Inflammation; Macrophages; Pain; Peptic Ulcer; Phenanthrenes; Piperidines; Stimulation, Chemical; Thromboxane B2

An inherited deficiency in the frataxin protein causes neurodegeneration of the dorsal root ganglia and Friedreich's ataxia (FA). Frataxin deficiency leads to oxidative stress and inflammatory changes in cell and animal models; however, the cause of the inflammatory changes, and especially what causes brain microglial activation is unclear. Here we investigated: 1) the mechanism by which frataxin deficiency activates microglia, 2) whether a brain-localized inflammatory stimulus provokes a greater microglial response in FA animal models, and 3) whether an anti-inflammatory treatment improves their condition. Intracerebroventricular administration of LPS induced higher amounts of microglial activation in the FA mouse model vs controls. We also observed an increase in oxidative damage in the form of 8-oxoguanine (8-oxo-G) and the DNA repair proteins MUTYH and PARP-1 in cerebellar microglia of FA mutant mice. We hypothesized that frataxin deficiency increases DNA damage and DNA repair genes specifically in microglia, activating them. siRNA-mediated frataxin knockdown in microglial BV2 cells clearly elevated DNA damage and the expression of DNA repair genes MUTYH and PARP-1. Frataxin knockdown also induced a higher level of PARP-1 in MEF cells, and this was suppressed in MUTYH-/- knockout cells. Administration of the PARP-1 inhibitor PJ34 attenuated the microglial activation induced by intracerebroventricular injection of LPS. The combined administration of LPS and angiotensin II provoke an even stronger activation of microglia and neurobehavioral impairment. PJ34 treatment attenuated the neurobehavioral impairments in FA mice. These results suggest that the DNA repair proteins MUTYH and PARP-1 may form a pathway regulating microglial activation initiated by DNA damage, and inhibition of microglial PARP-1 induction could be an important therapeutic target in Friedreich's ataxia.

Topics: Angiotensin II; Animals; Behavior, Animal; Cell Line; Cerebellum; Disease Models, Animal; DNA Damage; DNA Glycosylases; Female; Frataxin; Friedreich Ataxia; Gene Expression; Gene Expression Regulation; Gene Knockdown Techniques; Inflammation; Iron-Binding Proteins; Lipopolysaccharides; Mice; Microglia; Oxidative Stress; Phenanthrenes; Poly(ADP-ribose) Polymerases; RNA, Small Interfering

Poly (ADP-ribose) polymerases (PARPs) play an important role in a range of neurological disorders, however, the role of PARP in early brain injury after subarachnoid hemorrhage (SAH) remains unclear. This study was designed to explore the role and the potential mechanisms of PARP in early brain injury after SAH. Eighty-nine male SD rats were randomly divided into the Sham group, SAH+Vehicle group and SAH+PARP inhibitor (PJ34) group. An endovascular perforation model was used to induce SAH in rats. PJ34 (10mg/kg) or vehicle (0.9% NaCl) was intraperitoneally administered at 5min and 8h after SAH induction. Mortality, SAH grades, neurological function, evans blue extravasation, brain edema, immunofluorescence staining and western blotting were performed. PJ34 reduced BBB permeability and brain edema, improved neurological function and attenuated neuronal cell death in the rat model of SAH. Moreover, PJ34 inhibited the nuclear translocation of NF-κB, decreased the expression of the proinflammatory cytokines IL-1ß, IL-6 and TNF-α, reduced the expression of MMP-9, prevented the degradation of tight junction proteins, and decreased microglia activation. These data indicated that PARP inhibition through PJ34 might be an important therapeutic drug for SAH.

Topics: Animals; Cell Death; Claudin-5; Disease Models, Animal; Interleukin-1beta; Interleukin-6; Male; Matrix Metalloproteinase 9; Microglia; NF-kappa B; Occludin; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Rats; Rats, Sprague-Dawley; Signal Transduction; Subarachnoid Hemorrhage; Tumor Necrosis Factor-alpha

Our laboratory previously showed that sodium tanshinone IIA sulfonate (STS) inhibited store-operated Ca(2+) entry (SOCE) through store-operated Ca(2+) channels (SOCC) via downregulating the expression of transient receptor potential canonical proteins (TRPC), which contribute to the formation of SOCC (Wang J, Jiang Q, Wan L, Yang K, Zhang Y, Chen Y, Wang E, Lai N, Zhao L, Jiang H, Sun Y, Zhong N, Ran P, Lu W. Am J Respir Cell Mol Biol 48: 125-134, 2013). The detailed molecular mechanisms by which STS inhibits SOCE and downregulates TRPC, however, remain largely unknown. We have previously shown that, under hypoxic conditions, inhibition of protein kinase G (PKG) and peroxisome proliferator-activated receptor-γ (PPAR-γ) signaling axis results in the upregulation of TRPC (Wang J, Yang K, Xu L, Zhang Y, Lai N, Jiang H, Zhang Y, Zhong N, Ran P, Lu W. Am J Respir Cell Mol Biol 49: 231-240, 2013). This suggests that strategies targeting the restoration of this signaling pathway may be an effective treatment strategy for pulmonary hypertension. In this study, our results demonstrated that STS treatment can effectively prevent the hypoxia-mediated inhibition of the PKG-PPAR-γ signaling axis in rat distal pulmonary arterial smooth muscle cells (PASMCs) and distal pulmonary arteries. These effects of STS treatment were blocked by pharmacological inhibition or specific small interfering RNA knockdown of either PKG or PPAR-γ. Moreover, targeted PPAR-γ agonist markedly enhanced the beneficial effects of STS. These results comprehensively suggest that STS treatment can prevent hypoxia-mediated increases in intracellular calcium homeostasis and cell proliferation, by targeting and restoring the hypoxia-inhibited PKG-PPAR-γ signaling pathway in PASMCs.

Topics: Animals; Calcium Signaling; Cell Proliferation; Cells, Cultured; Cyclic GMP-Dependent Protein Kinases; Disease Models, Animal; Dose-Response Relationship, Drug; Hypertension, Pulmonary; Hypoxia; Male; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Phenanthrenes; PPAR gamma; Protein Kinase Inhibitors; Pulmonary Artery; Rats, Sprague-Dawley; RNA Interference; Time Factors; Transfection; TRPC Cation Channels; Vascular Remodeling

To investigate the therapeutic effect of triptolide (TP) on diabetic nephropathy (DN) in addition to its influence on helper T lymphocytes (Th) cells and monocytes/macrophages in rat models of DN.. Diabetes was induced in rats by feeding them high-fat diets and administering low-dose streptozotocin (STZ); subsequently, they were treated with TP (6, 12, or 24 mg/kg/day respectively) for 4 weeks. The general characteristics of the rats and their kidney weight to body weight ratio were observed. Liver and kidney function tests, routine blood tests, and 24 h urine protein tests were performed. Histological and ultrastructural pathologic changes in the kidneys were examined. Changes in the proportion of Th1/Th2 cells in peripheral blood and CD4(+) T cells were measured. The serum levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-4, and IL-10 were determined and the expression of these 4 cytokines in the kidneys was measured. Expression of the CD68 macrophage surface marker as well as that of phospho-nuclear factor kappa B (p-NF-κB), NF-κB, monocyte chemoattractant protein (MCP)-1, transforming growth factor (TGF)-β1, fibronectin (FN), IL-12, and signal transducer and activator of transcription-4 (STAT4) was evaluated in the kidneys.. Elevated urine micro-albumin (UMA) and renal histological and ultrastructural changes were observed after the induction of diabetes. DN was associated with delayed immune-inflammatory responses induced by up-regulation of the proportion of Th1 cells and increase of the pro-inflammatory cytokines IFN-γ and TNF-α secreted by Th1 cells. In addition, down-regulation of the proportion of Th2 cells and decrease of the anti-inflammatory cytokines IL-4 and IL-10 secreted by Th2 cells were observed. TP could improve DN by regulating the Th1/Th2 cell balance. Macrophage infiltration as well as expression of inflammatory and pro-fibrogenic factors significantly increased in the kidneys of diabetic rats, which were suppressed by TP with the improvement in the medium-dose TP group (12 mg/kg/d) being the most significant.. TP can improve DN by regulating the Th1/Th2 cell balance and by reducing macrophage infiltration as well as the expression of relevant inflammatory factors in the kidney.

Topics: Animals; Diabetic Nephropathies; Disease Models, Animal; Diterpenes; Epoxy Compounds; Immunosuppressive Agents; Macrophages; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley; Th1 Cells; Th2 Cells

Brain injury resulting from stroke or trauma can be exacerbated by the release of proinflammatory cytokines, proteases, and reactive oxygen species by activated microglia. The microglial activation resulting from brain injury is mediated in part by alarmins, which are signaling molecules released from damaged cells. The nuclear enzyme poly(ADP-ribose) polymerase-1 (PARP-1) has been shown to regulate microglial activation after brain injury, and here we show that signaling effects of the alarmin S100B are regulated by PARP-1. S100B is a protein localized predominantly to astrocytes. Exogenous S100B added to primary microglial cultures induced a rapid change in microglial morphology, upregulation of IL-1β, TNFα, and iNOS gene expression, and release of matrix metalloproteinase 9 and nitric oxide. Most, though not all of these effects were attenuated in PARP-1(-/-) microglia and in wild-type microglia treated with the PARP inhibitor, veliparib. Microglial activation and gene expression changes induced by S100B injected directly into brain were likewise attenuated by PARP-1 inhibition. The anti-inflammatory effects of PARP-1 inhibitors in acutely injured brain may thus be mediated in part through effects on S100B signaling pathways. GLIA 2016;64:1869-1878.

Topics: Alarmins; Animals; Animals, Newborn; Astrocytes; Benzimidazoles; Brain; Brain Injuries; Cells, Cultured; Cytokines; Disease Models, Animal; Gene Expression Regulation; Mice; Mice, Inbred C57BL; Mice, Knockout; Microglia; Nitric Oxide Synthase Type II; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Polysaccharides; S100 Calcium Binding Protein beta Subunit

The aim of this study was to detect the therapeutic effect of dioscin on collagen-induced arthritis (CIA). Mice model of CIA was induced by chicken collagen II and arthritis index was assessed. After suspension of dioscin (100mg/kg/d) or triptolide was intragastrically administered, the left paw swelling and body weight of each mouse were measured. Then tissue samples were assayed by histopathological analysis. The levels of Th1 and Th2 were detected by flow cytometry. The expression of p-STAT1, p-STAT4 and p-STAT6 was demonstrated by western blot analysis, and T-bet and GATA-3 expression was detected by RT-PCR. The paw swelling and arthritis index were decreased and body weight was increased in the high dose of dioscin group compared to the model group (P<0.05). Histopathological analysis revealed that the damage of synovium tissue in dioscin and triptolide group alleviated. The ratio of Th1/Th2 in the dioscin group (0.82±0.24) and triptolide group (0.99±0.44) was lower than that in the model group (1.84±0.70, P<0.05). Additionally, p-STAT4 expression was decreased, and both p-STAT6 and GATA3 expression was increased in the dioscin group than that in the model group (P<0.05). Dioscin might have some therapeutic effects on CIA through regulating the proportion of Th1/Th2 cells, which could reduce the expression of p-STAT4, increase the expression of p-STAT6 and GATA3 in the synovial tissue.

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Collagen Type II; Dioscorea; Diosgenin; Disease Models, Animal; Diterpenes; Epoxy Compounds; GATA3 Transcription Factor; Humans; Male; Mice; Mice, Inbred DBA; Phenanthrenes; STAT Transcription Factors; Synovial Membrane; Th1 Cells; Th1-Th2 Balance; Th2 Cells

The Nuclear factor (erythroid 2-derived)-like 2 (Nrf2) plays a key role in inflammation which is implicated in the pathophysiology of depression. The Nrf2 activators have antidepressant effects in animal models of depression. The present study was undertaken to examine whether TBE-31 [(±)-(4bS,8aR,10aS)-10a-ethynyl-4b,8,8-trimethyl-3,7-dioxo-3,4b,7,8,8a,9,10,10a-octahydrophenanthrene-2,6-dicarbonitrile] and MCE-1 [(±)-3-ethynyl-3-methyl-6-oxocyclohexa-1,4-dienecarbonitrile], the novel Nrf2 activators, could show antidepressant effects in inflammation model of depression. We found that TBE-31 and MCE-1 significantly potentiated nerve growth factor (NGF)-induced neurite outgrowth in PC12 cells, in a concentration dependent manner. The Nrf2 siRNA, but not negative control of siRNA, significantly blocked the potentiating effects of TBE-31 and MCE-1 on neurite outgrowth in PC12 cells. Furthermore, oral administration of TBE-31 or MCE-1 significantly attenuated an increase in serum levels of tumor necrosis factor-α (TNF-α) after administration of lipopolysaccharide (LPS: 0.5mg/kg). In the tail-suspension test and forced swimming test, oral administration of TBE-31 or MCE-1 significantly attenuated an increase in the immobility time after LPS (0.5mg/kg) administration. These findings suggest that the novel Nrf2 activators such as TBE-31 and MCE-1 might be potential therapeutic drugs for inflammation-related depression.

Topics: Animals; Antidepressive Agents; Behavior, Animal; Cyclohexanones; Depression; Disease Models, Animal; Inflammation; Male; Nerve Growth Factor; Neuronal Outgrowth; NF-E2-Related Factor 2; PC12 Cells; Phenanthrenes; Rats; Tumor Necrosis Factor-alpha

Triptolide (TPL), a main active ingredient of Tripterygium wilfordii has been shown to exert anti-inflammatory effect. The role of TPL on glomerular diseases remains unclear.. This study aims to investigate the potential anti-inflammatory effect of TPL in rats with membranous glomerulonephritis (MGN).. Our data showed that the pathological kidney damage was significantly alleviated by TPL treatment in MGN rats. We also found that MGN rats exhibited significantly higher (p < 0.01) level of inflammatory cytokines (TNF-α, IL-1β and MCP-1) than those in normal group, while these inflammatory cytokines levels were significantly reduced in TPL treatment group compared with model group. Additionally, we found that TPL treatment could significantly decrease the malondialdehyde (MDA) level while enhanced superoxide dismutase (SOD) activity. Meanwhile, we also found that IκB kinase inhibitor (IMD-0354) could significantly reduce the accumulation of inflammation damage and oxidative lesions. Furthermore, we observed that both TPL and IMD-0354 treatment could block IκBα degradation and suppress mRNA and protein level of nuclear factor (NF) -κB p65.. Together, all above results suggest that inflammatory response could be attenuated by TPL and this is partly due to the inhibition of NF- κB signaling pathway.

Topics: Animals; Anti-Inflammatory Agents; Disease Models, Animal; Diterpenes; Down-Regulation; Epoxy Compounds; Glomerulonephritis, Membranous; Inflammation; Male; NF-kappa B; Phenanthrenes; Rats; Rats, Sprague-Dawley; Signal Transduction

We have reported that triptolide inhibited pulmonary inflammation in patients with steroid-resistant asthma. In the present study, we investigated whether suppresses airway remodeling and goblet cell hyperplasia, studied the mechanism of triptolide on mucin5ac (Muc5ac) expression in a murine model of asthma.. BALB/c mice were sensitized to intraperitoneal ovalbumin (OVA) followed by repetitive ovalbumin challenge for 6 weeks. Treatments included triptolide (40 μg/kg) and dexamethasone (2mg/kg). The area of bronchial airway (WAt/Pbm), smooth muscle (WAm/Pbm) and mucus index were assessed 24h after the final OVA challenge. Levels of Muc5ac were assessed by ELISA, immunohistology and real-time PCR. Western blot was performed to analyze the phosphorylation of NF-κB p65.. Triptolide and dexamethasone significantly reduced allergen-induced increases in the thickness of bronchial airway, smooth muscle and goblet cell hyperplasia. Levels of lung Muc5ac and Muc5ac mRNA were significantly reduced in mice treated with triptolide and dexamethasone. Phosphorylation of NF-κB p65 was significantly reduced in mice treated with triptolide and dexamethasone.. Triptolide may inhibit airway goblet cell hyperplasia and Muc5ac expression in asthmatic mice via NF-κB. It may be a potential drug for the treatment of patients with severe asthma.

Topics: Airway Remodeling; Animals; Asthma; Blotting, Western; Bronchi; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Goblet Cells; Hyperplasia; Hypertrophy; Mice, Inbred BALB C; Mucin 5AC; Mucus; NF-kappa B; Phenanthrenes; Phosphorylation; RNA, Messenger; Transcription Factor RelA

Preeclampsia is a disorder of pregnancy with a significant impact on maternal and fetal health. The complexity of this multifactorial condition has precluded development of effective therapies and, although many potential pathways have been investigated, the etiology still requires clarification. Our group has investigated the scavenger lectin-like oxidized LDL (LOX-1) receptor, which may respond to factors released from the distressed placenta that contribute to the vascular pathologies observed in preeclampsia. Given the known beneficial effects of sodium tanshinone IIA sulfonate (STS; a component of Salvia miltiorrhiza) on vasodilation, reduction of oxidative stress, and lipid profiles, we have investigated its role as a potential treatment strategy. We hypothesized that STS would improve vascular endothelial function and, combined with a reduction in oxidative stress, would improve pregnancy outcomes in a rat model of preeclampsia (reduced uteroplacental perfusion pressure, RUPP). We further hypothesized this may occur via the action of STS on the LOX-1 and/or platelet-activating factor (PAF) receptor axes. The RUPP model increased maternal blood pressure, vascular oxidative stress, and involvement of the vascular PAF receptor. Treatment with STS during pregnancy decreased both oxidative stress and involvement of the PAF receptor; however, it also increased involvement of the LOX-1 receptor, which is in line with the concept that scavenger receptors, such as LOX-1 and PAF, are upregulated in response to ligand binding and/or under pathological conditions. In this model of preeclampsia, however, the vascular actions of STS did not lead to improvements in pregnancy outcome such as fetal biometrics or maternal blood pressure.

Topics: Animals; Blood Pressure; Disease Models, Animal; Endothelium, Vascular; Female; Lipoproteins, LDL; Oxidative Stress; Phenanthrenes; Placenta; Pre-Eclampsia; Pregnancy; Rats, Sprague-Dawley; Vasodilation

Estrogen receptor (ER) is a major therapeutic target for the treatment of breast cancer, because of the crucial role of estrogen signaling deregulation in the development and progression of breast cancer. In this study, we report the identification of a novel ERα binding compound, cryptotanshinone (CPT), by screening the CADD database. We also show that CPT effectively inhibits estrogen-induced ER transactivation and gene expression of ER target genes. Furthermore, we showed that CPT suppressed breast cancer cell growth mainly in an ERα dependent manner. Finally, we confirmed the potential therapeutic efficiency of CPT using xenograft experiments in vivo. Taken together, our results describe a novel mechanism for the anticancer activity of CPT and provide supporting evidence for its use as a potential therapeutic agent to treat patients with ERα positive breast cancer.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; Estrogen Receptor alpha; Female; Gene Expression Regulation, Neoplastic; Humans; Mice; Models, Molecular; Molecular Docking Simulation; Phenanthrenes; Protein Conformation; Receptors, Estrogen; Signal Transduction; Transcription, Genetic; Tumor Burden; Xenograft Model Antitumor Assays

In this study, we studied the effect of triptolide (TPL) on locomotor function in rats with spinal cord injury. A total of 40 rats were studied after dividing them in two major groups, one was experimental group denoted as TPL group while other was control group denoted as PBS group. Each group was subdivided in four subgroups having five rats each (n = 5). TPL was given intraperitonially at the rate of 5 mg/kg/day in TPL group while PBS was given at the same time interval in the same manner in control group for comparison. A reduction in the cavity area of tissue sections was observed by bright field microscopy from 0.22 ± 0.05 to 0.12 ± 0.05 mm(2) in experimental group after 28 days of treatment while BBB score also improved from 1 to 5 after 14 days of treatment. SPSS software, one way ANOVA, was used for recording statistical analysis and values were expressed as mean ± SEM where P value of <0.01 was considered significant. The expression of I-kBα and NF-kB p65 was also studied using western blotting and after recording optical density (OD) values of western blots. It was observed that treatment with TPL significantly reduced the expression of these factors after 28 days of treatment compared with controls.

Topics: Animals; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; I-kappa B Proteins; Inflammation; Motor Activity; Phenanthrenes; Rats; Rats, Sprague-Dawley; Recovery of Function; Spinal Cord Injuries; Time Factors

We previously showed that activity of the large conductance calcium-activated potassium (Big-K; BK) channels is suppressed in 3xTg Alzheimer disease (AD) model mice. However, its behavioral significance is not known. In the present report, ventricular injection of the BK channel activator isopimaric acid (ISO) was conducted to examine whether BK channel activation ameliorates cognition in 3xTg mice. The novel object recognition (NOR) test revealed that chronic injection of ISO improved non-spatial memory in 3xTg mice. In the Morris water maze, the probe test demonstrated an improved spatial memory after ISO injection. Electrophysiological underpinnings of the ISO effect were then examined in slices obtained from the mice after behavior. At hippocampal CA1 synapses, the basic synaptic transmission was abnormally elevated and long-term potentiation (LTP) was partially suppressed in 3xTg mice. These were both recovered by ISO treatment. We then confirmed suppressed BK channel activity in 3xTg mice by measuring the half-width of evoked action potentials. This was also recovered by ISO treatment. We previously showed that the recovery of BK channel activity accompanies reduction of neuronal excitability in pyramidal cells. Here again, pyramidal cell excitability, as assessed by calculating the frequency of evoked spikes, was elevated in the 3xTg mouse and was normalized by ISO. ELISA experiments demonstrated an ISO-induced reduction of Aβ1-42 content in hippocampal tissue in 3xTg mice. The present study thus suggests a potential therapeutic utility of BK channel activators for AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; CA1 Region, Hippocampal; Carboxylic Acids; Charybdotoxin; Cognition Disorders; Disease Models, Animal; Drug Delivery Systems; Gene Expression Regulation; Humans; In Vitro Techniques; Large-Conductance Calcium-Activated Potassium Channels; Long-Term Potentiation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neurotoxins; Patch-Clamp Techniques; Peptide Fragments; Phenanthrenes; Recognition, Psychology

Poly(ADP-ribose) polymerase (PARP) is activated by oxidative stress and plays an important role in traumatic brain injury (TBI). The objective of this study was to investigate whether PARP activation participated in the blood-brain barrier (BBB) disruption and edema formation in a mouse model of controlled cortical impact (CCI). N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide (PJ34) (10 mg/kg), a selective PARP inhibitor, was administered intraperitoneally at 5 min and 8 h after experimental CCI. After 6 h and 24 h of CCI, the permeability of the cortical BBB was determined after Evans Blue administration. The water content of the brain was also measured. Treatment with PJ34 markedly attenuated the permeability of the BBB and decreased the brain edema at 6 h and 24 h after CCI. Our data showed the up-regulation of nuclear factor-κB in cytosolic fractions and nuclear fractions in the injured cortex, and these changes were reversed by PJ34. Moreover, PJ34 significantly lessened the activities of myeloperoxidase and the levels of matrix metalloproteinase-9, enhanced the levels of occludin, laminin, collagen IV and integrin β1, reduced neurological deficits, decreased the contusion volume, and attenuated the necrotic and apoptotic neuronal cell death. These data suggest the protective effects of PJ34 on BBB integrity and cell death during acute TBI.

Topics: Animals; Blood-Brain Barrier; Brain; Brain Edema; Brain Injuries; Capillary Permeability; Cell Death; Disease Models, Animal; Male; Matrix Metalloproteinase 9; Mice, Inbred BALB C; Neurons; Neuroprotective Agents; NF-kappa B; Occludin; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Random Allocation

The use of triptolide (TP) is limited by its poor water solubility and severe toxicity. In this study, we developed an active drug delivery system (TP-loaded nanoparticles) that could help improve the water solubility of TP and decrease its toxicity. Then, we investigated whether TP-loaded nanoparticles could be used to establish a novel premature ovarian insufficiency mouse model. The mice treated with TP-loaded nanoparticles for 35 days displayed normal growth, decreased serum antimullerian hormone, prominent ovarian fibrosis and vacuolar changes, fewer follicles and corpus lutea, increased collapsed oocytes and follicle apoptosis, and sterility. In conclusion, this model appears to show the reproductive characteristics associated with premature ovarian insufficiency in women and will allow us to study the mechanism of the effects of traditional Chinese medicine on gonadal toxicity.

Topics: Animals; Apoptosis; Disease Models, Animal; Diterpenes; Drug Delivery Systems; Epoxy Compounds; Female; Fertility; Follicle Stimulating Hormone; In Situ Nick-End Labeling; Mice; Mice, Inbred C57BL; Nanoparticles; Phenanthrenes; Primary Ovarian Insufficiency

Macrophages are important therapeutic targets for various disorders, including infectious diseases, inflammatory diseases, metabolic diseases, and cancer. In this study, we report a novel oral delivery system for the targeted delivery of anti-inflammatory therapeutics to macrophages. Using this formulation, the model drug tylophorine malate (NK007) was tightly incorporated inside beta-glucan particle shells by the formation of colloidal particles with chitosan, tripolyphosphate, and alginate via electrostatic interactions. This formulation specifically delivered NK007 to macrophages in vivo after oral gavage and effectively cured colitis in the dextran sulfate sodium-induced murine colitis model, highlighting the utility of beta-glucan particles as an oral anti-inflammation drug delivery system by targeting macrophages. In this work, NK007 was selected as the model drug. However, this novel oral carrier system has the potential to be applied as a platform for the treatment of many other diseases for which macrophages are the therapeutic targets.

Topics: Alginates; Alkaloids; Animals; Anti-Inflammatory Agents; Chemistry, Pharmaceutical; Chitosan; Colitis; Colon; Dextran Sulfate; Disease Models, Animal; Drug Delivery Systems; Glucuronic Acid; Hexuronic Acids; Indolizines; Macrophages; Mice; Pharmaceutical Preparations; Phenanthrenes

Transcranial magnetic stimulation (TMS) is fragmentarily reported to be beneficial to Alzheimer's patients. Its underlying mechanism was investigated. TMS was applied at 1, 10 or 15 Hz daily for 4 weeks to young Alzheimer's disease model mice (3xTg), in which intracellular soluble amyloid-β is notably accumulated. Hippocampal long-term potentiation (LTP) was tested after behavior. TMS ameliorated spatial learning deficits and enhanced LTP in the same frequency-dependent manner. Activity of the large conductance calcium-activated potassium (Big-K; BK) channels was suppressed in 3xTg mice and recovered by TMS frequency-dependently. These suppression and recovery were accompanied by increase and decrease in cortical excitability, respectively. TMS frequency-dependently enhanced the expression of the activity-dependently expressed scaffold protein Homer1a, which turned out to enhance BK channel activity. Isopimaric acid, an activator of the BK channel, magnified LTP. Amyloid-β lowering was detected after TMS in 3xTg mice. In 3xTg mice with Homer1a knocked out, amyloid-β lowering was not detected, though the TMS effects on BK channel and LTP remained. We concluded that TMS facilitates BK channels both Homer1a-dependently and -independently, thereby enhancing hippocampal LTP and decreasing cortical excitability. Reduced excitability contributed to amyloid-β lowering. A cascade of these correlated processes, triggered by TMS, was likely to improve learning in 3xTg mice.

Topics: Alzheimer Disease; Animals; Carboxylic Acids; Carrier Proteins; Disease Models, Animal; Hippocampus; Homer Scaffolding Proteins; Large-Conductance Calcium-Activated Potassium Channels; Long-Term Potentiation; Male; Maze Learning; Membrane Transport Modulators; Mice, 129 Strain; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Neurons; Phenanthrenes; Tissue Culture Techniques; Transcranial Magnetic Stimulation

Friedreich's ataxia (FRDA) is an autosomal recessive disorder, caused by reduced levels of the protein frataxin. This protein is located in the mitochondria, where it functions in the biogenesis of iron-sulphur clusters (ISCs), which are important for the function of the mitochondrial respiratory chain complexes. Moreover, disruption in iron biogenesis may lead to oxidative stress. Oxidative stress can be the cause and/or the consequence of mitochondrial energy imbalance, leading to cell death. Fibroblasts from two FRDA mouse models, YG8R and KIKO, were used to analyse two different categories of protective compounds: deuterised poly-unsaturated fatty acids (dPUFAs) and Nrf2-inducers. The former have been shown to protect the cell from damage induced by lipid peroxidation and the latter trigger the well-known Nrf2 antioxidant pathway. Our results show that the sensitivity to oxidative stress of YG8R and KIKO mouse fibroblasts, resulting in cell death and lipid peroxidation, can be prevented by d4-PUFA and Nrf2-inducers (SFN and TBE-31). The mitochondrial membrane potential (ΔΨm) of YG8R and KIKO fibroblasts revealed a difference in their mitochondrial pathophysiology, which may be due to the different genetic basis of the two models. This suggests that variable levels of reduced frataxin may act differently on mitochondrial pathophysiology and that these two cell models could be useful in recapitulating the observed differences in the FRDA phenotype. This may reflect a different modulatory effect towards cell death that will need to be investigated further.

Topics: Animals; Antioxidants; Cell Death; Cells, Cultured; Disease Models, Animal; Fibroblasts; Frataxin; Friedreich Ataxia; Iron; Iron-Binding Proteins; Lipid Peroxidation; Membrane Potential, Mitochondrial; Mice; Mice, Inbred C57BL; Mitochondria; Oxidative Stress; Phenanthrenes; Phenotype

The activation of NOD-like receptor (NLR) family, pyrin-domain containing 3 (NLRP3) inflammasome has now been proven to have a close connection with myocardial ischemia (MI) during acute phase, but the mechanisms are not completely clear. This study investigated the role of NLRP3 inflammasome in pathogenesis of MI injury including inflammation and lipid accumulation, as well as the effects of sodium tanshinone IIA sulfonate (STS) and diltiazem hydrochloride (DI).. Occlusion of left anterior descending (LAD) in canines was employed to induce MI. STS and DI were given intravenously 15 min after LAD occlusion. Cardiac function, inflammation and lipid levels, as well as related signaling pathways were determined.. MI induced in Beagle dog was characterized by elevated ST-segment and increased CK-MB level in serum. Cardiac NLRP3 inflammasome was activated with elevated myocardial IL-1β and IL-18 concentrations mediated by ROS over-production and TXNIP over-expression in MI dogs. Additionally, pro-inflammatory cytokines induced impairment of cardiac JAK2-STAT3 inflammatory pathway and insulin signaling pathway in this model, resulting in down-regulation of cardiac PPAR-α expression, subsequently causing lipid metabolism disorders characterized by elevation of myocardial lipid concentrations. These abnormalities were attenuated by the treatment of STS and DI.. These data firstly demonstrated that cardiac NLRP3 inflammasome activation driven by cardiac ROS over-production and TXNIP up-expression resulted in impairment of the JAK2-STAT3 and insulin signaling pathways, leading to disorder of lipid metabolism in myocardial ischemic dogs through PPAR-α over-expression. STS and DI might target cardiac NLRP3 inflammasome in preventing MI injury.

Topics: Animals; Biomarkers; Carrier Proteins; Coronary Occlusion; Coronary Vessels; Creatine Kinase, MB Form; Disease Models, Animal; Dogs; Drugs, Chinese Herbal; Electrocardiography; Inflammasomes; Janus Kinase 2; Lipid Metabolism; Male; Myocardial Ischemia; Phenanthrenes; PPAR alpha; Reactive Oxygen Species; Signal Transduction; STAT3 Transcription Factor

A gene encoding a functionally unknown protein that is specifically expressed in the thyroidectomized chicken fatty liver and has a predicted amino acid sequence similar to that of NAD(P)H-dependent carbonyl reductase was overexpressed in Escherichia coli; its product was purified and characterized. The expressed enzyme was an NAD(P)H-dependent broad substrate specificity carbonyl reductase and was inhibited by arachidonic acid at 1.5 μm. Enzymological characterization indicated that the enzyme could be classified as a cytosolic-type carbonyl reductase. The enzyme's 3D structure was determined using the molecular replacement method at 1.98 Å resolution in the presence of NADPH and ethylene glycol. The asymmetric unit consisted of two subunits, and a noncrystallographic twofold axis generated the functional dimer. The structures of the subunits, A and B, differed from each other. In subunit A, the active site contained an ethylene glycol molecule absent in subunit B. Consequently, Tyr172 in subunit A rotated by 103.7° in comparison with subunit B, which leads to active site closure in subunit A. In Y172A mutant, the Km value for 9,10-phenanthrenequinone (model substrate) was 12.5 times higher than that for the wild-type enzyme, indicating that Tyr172 plays a key role in substrate binding in this carbonyl reductase. Because the Tyr172-containing active site lid structure (Ile164-Gln174) is not conserved in all known carbonyl reductases, our results provide new insights into substrate binding of carbonyl reductase. The catalytic properties and crystal structure revealed that thyroidectomized chicken fatty liver carbonyl reductase is a novel enzyme.

Topics: Aldehyde Reductase; Aldo-Keto Reductases; Amino Acid Sequence; Amino Acid Substitution; Animals; Biocatalysis; Catalytic Domain; Chickens; Databases, Protein; Disease Models, Animal; Fatty Liver; Gene Expression Regulation, Enzymologic; Hypothyroidism; Liver; Molecular Sequence Data; Mutant Proteins; NADP; Phenanthrenes; Protein Conformation; Protein Stability; Protein Subunits; Recombinant Proteins; Tyrosine

IgA nephropathy (IgAN) is the finding of immune deposits predominantly containing polymeric IgA in the glomerular mesangium on renal biopsy. Recently studies show that inflammation may involve in the progression of renal glomerulosclerosis and tubulointerstitial scarring in IgAN. This study was designed to evaluate the renoprotective effect of triptolide on IgAN rat model. IgAN was induced in Sprague-Dawley rats by oral and intravenous immunization with BSA for 12 weeks. Rats were treated with triptolide (200 μg/kg/d intragastrically) from 12 to 28 weeks. At Week 28, the rats was sacrificed, kidneys and blood samples were collected for further analysis. Our data shown that IgAN rat model showed marked deterioration of proteinuria together with higher levels of the urine protein:creatinine ratio compared to the normal control. Animals that underwent intermittent exposure to triptolide treatment exhibited significant improvements in the functional parameters without severe side effects. Rats developing IgAN had profound mesangial proliferation and mesangial expansion, intense and diffuse glomerular IgA deposition, while triptolide treatment significantly attenuated it. We also observed that treatment with triptolide significantly decreases serum levels of IL-1β and IL-18, and may exerted anti-inflammatory effects by down-regulating NLRP3 and TLR4 expression. Our study clearly demonstrated that triptolide prevents IgAN progression via an amelioration of inflammasome-mediated proinflammatory cytokine production, thus brought a light of hope for treatment of IgAN.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrier Proteins; Disease Models, Animal; Diterpenes; Epoxy Compounds; Gene Expression Regulation; Glomerulonephritis, IGA; Interleukin-18; Interleukin-1beta; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Phenanthrenes; Rats; Rats, Sprague-Dawley; Toll-Like Receptor 4

The effects of sodium tanshinone IIA sulfonate (STS) on coronary no-reflow (CNR) relevant to microvascular obstruction (MVO) remain unknown. Studies had shown that fibrinogen-like protein 2 (FGL2) expressed in microvascular endothelial cells (MECs) is a key mediator in MVO. Thus, we aimed to elucidate the roles of STS in CNR and relations between STS and FGL2.. Myocardial ischemia/reperfusion was selected to represent CNR model. The no-reflow zone and infarct area were assessed using Thioflavin S and TTC staining, and cardiac functional parameters were detected using echocardiography. Western blot was used to detected FGL2 level, fibrin level, protease-activated receptor-1 (PAR-1) activation and inflammation cells infiltration. FGL2 and inflammation cells were also identified by IHC. Microthrombus was detected by Carstairs' and MSB staining. We also detected the roles of STS on FGL2 expression, thrombin generation, phospho-Akt and NF-κB levels in MECs.. Upon treatment with STS in CNR model, the no-reflow and infarct areas decreased significantly and cardiac function improved. The FGL2 expression was inhibited by STS in vivo as well as in vitro with thrombin generation inhibition. In addition, STS up-regulates Akt phosphorylation and suppressed NF-κB expression in activated MECs. Furthermore, fibrin deposition, PAR-1 activation and inflammatory response were inhibited with STS administration in CNR model.. Our results displayed a novel pharmacological action of STS on CNR. STS is able to ameliorate CNR through inhibition of FGL2 expression mediated by Akt and NF-κB pathways as well as prevention of MVO by suppressing fibrin deposition and inflammation.

Topics: Animals; Coronary Circulation; Disease Models, Animal; Down-Regulation; Endothelial Cells; Fibrin; Fibrinogen; Male; No-Reflow Phenomenon; Phenanthrenes; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Receptor, PAR-1; Reperfusion Injury; Signal Transduction

To study the effects of triptolide, a Chinese herb extract, on retinal ganglion cells (RGCs) in a rat model of chronic glaucoma.. Eighty Wistar rats were randomly divided into triptolide group (n=40) and normal saline (NS) group (n=40). Angle photocoagulation was used to establish the model of glaucoma, with right eye as laser treated eye and left eye as control eye. Triptolide group received triptolide intraperitoneally daily, while NS group received NS. Intraocular pressure (IOP), anti-CD11b immunofluorescent stain in retina and optic nerve, RGCs count with Nissel stain and microglia count with anti-CD11b immunofluorescence stain in retina flat mounts, retinal tumor necrosis factor (TNF)-α mRNA detection by reverse transcription-polymerase chain reaction, and double immunofluorescent labeling with anti-TNF-α and anti-CD11b in retinal frozen section were performed.. Mean IOP of the laser treated eyes significantly increased 3 weeks after photocoagulation (P<0.05), with no statistical difference between the two groups (P>0.05). RGCs survival in the laser treated eyes was significantly improved in the triptolide group than the NS group (P<0.05). Microglia count in superficial retina of the laser treated eyes was significantly less in the triptolide group (30.40±4.90) than the NS group (35.06±7.59) (P<0.05). TNF-α mRNA expression in the retina of the laser treated eyes in the triptolide group decreased by 60% compared with that in the NS group (P<0.01). The double immunofluorescent labeling showed that TNF-α was mainly distributed around the microglia.. Triptolide improved RGCs survival in this rat model of chronic glaucoma, which did not depend on IOP decrease but might be exerted by inhibiting microglia activities and reducing TNF-α secretion.

Topics: Animals; Cell Survival; Disease Models, Animal; Diterpenes; Epoxy Compounds; Glaucoma; Injections, Intraperitoneal; Intraocular Pressure; Microglia; Phenanthrenes; Rats; Rats, Wistar; Retinal Ganglion Cells; Tumor Necrosis Factor-alpha

Although the exact cause of Alzheimer's disease (AD) remains elusive, mounting evidence continues to support the involvement of neuroinflammation in the development of AD. Triptolide isolated from the herb Tripterygium wilfordii Hook F has anti-inflammatory and immunosuppressive activities. In this study, we observed the effects of triptolide on dendritic spines of hippocampal neurons in model rats with AD. Thirty male SD rats were randomly divided into control group, AD model group and triptolide-treated group. The AD model group was made with bilateral microinjection of aggregated beta-amyloid protein (Aβ)1-40 into hippocampus in rats and the control group rats were injected with normal saline in the same way. The triptolide-treated group rats were administered triptolide intraperitoneally for 30 days after microinjection of aggregated Aβ1-40 into hippocampus. Dendritic morphology of hippocampal neurons in each group was analyzed using Golgi staining and ImageJ software. Our data showed that the total number of intersection points of dendrites and spine density in hippocampal neurons in the AD model group were decreased as compared with the control group. However, the total number of intersection points of dendrites and spine density in hippocampal neurons in the triptolide-treated group were increased as compared with the AD model group. Our results indicate that triptolide can alleviate the degeneration of dendritic spines in hippocampal neurons in model rats with AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Dendritic Spines; Disease Models, Animal; Diterpenes; Epoxy Compounds; Hippocampus; Immunosuppressive Agents; Male; Nerve Degeneration; Peptide Fragments; Phenanthrenes; Rats; Rats, Sprague-Dawley

Traumatic brain injury (TBI) causes neuronal cell death as well as microglial activation and related neurotoxicity that contribute to subsequent neurological dysfunction. Poly (ADP-ribose) polymerase (PARP-1) induces neuronal cell death through activation of caspase-independent mechanisms, including release of apoptosis inducing factor (AIF), and microglial activation. Administration of PJ34, a selective PARP-1 inhibitor, reduced cell death of primary cortical neurons exposed to N-Methyl-N'-Nitro-N-Nitrosoguanidine (MNNG), a potent inducer of AIF-dependent cell death. PJ34 also attenuated lipopolysaccharide and interferon-γ-induced activation of BV2 or primary microglia, limiting NF-κB activity and iNOS expression as well as decreasing generation of reactive oxygen species and TNFα. Systemic administration of PJ34 starting as late as 24 h after controlled cortical impact resulted in improved motor function recovery in mice with TBI. Stereological analysis demonstrated that PJ34 treatment reduced the lesion volume, attenuated neuronal cell loss in the cortex and thalamus, and reduced microglial activation in the TBI cortex. PJ34 treatment did not improve cognitive performance in a Morris water maze test or reduce neuronal cell loss in the hippocampus. Overall, our data indicate that PJ34 has a significant, albeit selective, neuroprotective effect after experimental TBI, and its therapeutic effect may be from multipotential actions on neuronal cell death and neuroinflammatory pathways.

Topics: Animals; Blotting, Western; Brain Injuries; Cell Death; Disease Models, Animal; Enzyme Inhibitors; Male; Mice; Mice, Inbred C57BL; Microglia; Neurons; Neuroprotective Agents; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Rats, Sprague-Dawley; Recovery of Function

Pancreatic adenocarcinoma is the fourth leading cause for cancer-related mortality with a survival rate of less than 5%. Late diagnosis and lack of effective chemotherapeutic regimen contribute to these grim survival statistics. Relapse of any tumor is largely attributed to the presence of tumor-initiating cells (TIC) or cancer stem cells (CSC). These cells are considered as hurdles to cancer therapy as no known chemotherapeutic compound is reported to target them. Thus, there is an urgent need to develop a TIC-targeted therapy for pancreatic cancer.. We isolated CD133(+) cells from a spontaneous pancreatic ductal adenocarcinoma mouse model and studied both surface expression, molecular markers of pancreatic TICs. We also studied tumor initiation properties by implanting low numbers of CD133(+) cells in immune competent mice. Effect of Minnelide, a drug currently under phase I clinical trial, was studied on the tumors derived from the CD133(+) cells.. Our study showed for the first time that CD133(+) population demonstrated all the molecular markers for pancreatic TIC. These cells initiated tumors in immunocompetent mouse models and showed increased expression of prosurvival and proinvasive proteins compared to the CD133(-) non-TIC population. Our study further showed that Minnelide was very efficient in downregulating both CD133(-) and CD133(+) population in the tumors, resulting in a 60% decrease in tumor volume compared with the untreated ones.. As Minnelide is currently under phase I clinical trial, its evaluation in reducing tumor burden by decreasing TIC as well as non-TIC population suggests its potential as an effective therapy.

Topics: AC133 Antigen; Animals; Antigens, CD; Antigens, Surface; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cell Transformation, Neoplastic; Disease Models, Animal; Diterpenes; Epoxy Compounds; Gene Expression; Glycoproteins; Immunophenotyping; Mice; Mice, Transgenic; Neoplastic Stem Cells; NF-kappa B; Organophosphates; Pancreatic Neoplasms; Peptides; Phenanthrenes; Phenotype

Cryptotanshinone is one of the compounds extracted from the root of Salvia miltiorrhiza Bunge. Unlike other tanshinones, only a small number of studies have focused on cryptotanshinone for medical treatment. In the present study, the A549 lung cancer cell line and xenograft models of human lung tumors were used to assess the anti-cancer effect of cryptotanshinone. The effect of cryptotanshinone on human lung cancer, including growth inhibition, cell cycle arrest and apoptosis factors, were identified in vitro, and inhibition of tumor formation, improvement of body condition as well as pathological apoptotic effects were detected in vivo. These results suggested that cryptotanshinone is a potential drug for the treatment and prevention of human lung cancer.

Topics: Animals; Apoptosis; Body Weight; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Transformation, Neoplastic; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Humans; Lung Neoplasms; Mice; Phenanthrenes; Tumor Burden; Tumor Stem Cell Assay; Xenograft Model Antitumor Assays

Glioblastoma multiforme (GBM) is a lethal solid cancer in adults. Temozolomide (TMZ) is a first-line chemotherapeutic agent but the efficacy is limited by intrinsic and acquired resistance in GBM. Triptolide (TPL), a derivative from traditional Chinese medicine, demonstrated anti-tumor activity. In this study, we explored the interaction of TPL and TMZ in glioma-initiating cells (GICs) and the potential mechanism. A GIC line (GIC-1) was successfully established. Cell viability of GIC-1 after treatment was measured using a CCK-8 assay. The interaction between TPL and TMZ was calculated from Chou-Talalay equations and isobologram. Self-renewal was evaluated with tumor sphere formation assay. Apoptosis was assessed with flow cytometry and western blot. Luciferase assay was employed to measure NF-κB transcriptional activity. The expression of NF-κB downstream genes, NF-κB nuclear translocalization and phoshorylation of IκBα and p65 were evaluated using western blot. We found that GIC-1 cells were resistant to TMZ, with the expected IC50 of 705.7 μmol/L. Co-treatment with TPL yielded a more than three-fold dose reduction of TMZ. TPL significantly increased the percentage of apoptotic cells and suppressed the tumor sphere formation when combined with TMZ. Phosphorylation of IκBα and p65 coupled with NF-κB nuclear translocalization were notably inhibited after a combined treatment. Co-incubation synergistically repressed NF-κB transcriptional activity and downstream gene expression. TPL sensitizes GICs to TMZ by synergistically enhancing apoptosis, which is likely resulting from the augmented repression of NF-κB signaling. TPL is therefore a potential chemosensitizer in the treatment of GBM.

Topics: Animals; Antineoplastic Agents, Alkylating; Apoptosis; Cell Line, Tumor; Cell Transformation, Neoplastic; Dacarbazine; Disease Models, Animal; Diterpenes; Drug Synergism; Epoxy Compounds; Glioma; Humans; Mice; Neoplasm Transplantation; NF-kappa B; Phenanthrenes; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; Stimulation, Chemical; Temozolomide; Transcription, Genetic; X-Linked Inhibitor of Apoptosis Protein

Triptolide (TPL) is a major active component isolated from the natural herb Tripteryglum wilfordii Hook. F. It has proved to possess a variety of pharmacological effects including anti-inflammatory and antitumor activities. The aim of the present study is to explore the efficiency of combination therapy with TPL and oxaliplatin (OXA) and identify the in vitro and in vivo cytotoxicity on colon cancer lines and mice model. Cell viability was measured by MTT assay and cell apoptosis rate was analyzed by FACS assay after treatment with TPL and OXA alone, and TPL combined with OXA in colon cancer cell line SW480. The results demonstrated that combination therapy of TPL and OXA could effectively inhibit the proliferation of colon cancer cell line SW480 and induce cell apoptosis of colon cancer cells. It was partly induced by inhibiting nuclear translocation of β-catenin and the expression of the target genes in cell cycle, which was detected by western blotting and real-time PCR. Moreover, in nude mice model, tumor growth was significantly suppressed in the group treated with TPL in combination with OXA. There was no obvious cytotoxity in mice analyzed by normal blood test and liver and kidney toxicity test. In conclusion, our result revealed that the combination therapy with TPL and OXA exerted synergistic antitumor effects at low concentration in colon cancer cells, with less cytotoxity, which exhibited high potency for clinical applications.

Topics: Animals; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; beta Catenin; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Disease Models, Animal; Diterpenes; Drug Synergism; Epoxy Compounds; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Male; Mice; Molecular Structure; Organoplatinum Compounds; Oxaliplatin; Phenanthrenes; Protein Transport

Triptolide is one of the main active components in the Chinese herb Tripterygium wilfordii Hook F, which has been demonstrated to possess anti‑inflammatory properties. The aim of this study was to investigate the effects of triptolide on lipopolysaccharide (LPS)‑induced acute lung injury (ALI) in mice and to explore the possible mechanisms. Mice were administered LPS intranasally to induce lung injury, and triptolide was administered intraperitoneally 1 h prior to the LPS challenge. Triptolide‑treated mice exhibited significantly reduced levels of leukocytes, myeloperoxidase activity and edema of the lung, as well as tumour necrosis factor‑α, interleukin (IL)‑1β and IL‑6 production in the bronchoalveolar lavage fluid compared with LPS‑treated mice. Additionally, western blot analysis showed that triptolide inhibited the LPS‑induced phosphorylation of nuclear factor of κ light polypeptide gene enhancer in B cells inhibitor‑α and nuclear factor κ‑light‑chain‑enhancer of activated B cells‑p65 (NF‑κB p65) and the expression of Toll‑like receptor 4 (TLR4). In conclusion, the results from the present study suggest that the anti‑inflammatory effect of triptolide against LPS‑induced ALI may be due to its ability to inhibit the TLR4‑mediated NF‑κB signalling pathway. Triptolide may therefore be a promising potential therapeutic agent for ALI treatment, which may ultimately aid the clinical therapy for patients with ALI.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents, Non-Steroidal; Bronchoalveolar Lavage Fluid; Chemokines; Disease Models, Animal; Diterpenes; Epoxy Compounds; I-kappa B Proteins; Interleukin-1beta; Interleukin-6; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; NF-kappa B; NF-KappaB Inhibitor alpha; Phenanthrenes; Phosphorylation; Signal Transduction; Tumor Necrosis Factor-alpha

It has been shown that triptolide has beneficial effects in the treatment of neuropathic pain, but its effects on bone cancer pain (BCP) remain unclear. In this study, we aimed to explore the potential role of spinal regulated activation of normal T cell expressed and secreted (RANTES) in the antinociceptive effects of triptolide on BCP. A BCP model was induced by injecting Walker 256 mammary gland carcinoma cells into the intramedullary space of rat tibia. Intrathecal administration of triptolide (0.5, 1, 2 μg) could dose-dependently alleviate mechanical hyperalgesia and spontaneous pain. In addition, there were also concomitant decreases in RANTES mRNA and protein expression levels in spinal dorsal horn. These results suggest that the antinociceptive effects of triptolide are related with inhibition of spinal RANTES expression in BCP rats. The findings of this study may provide a promising drug for the treatment of BCP.

Topics: Analgesics; Animals; Blotting, Western; Bone Neoplasms; Carcinoma 256, Walker; Chemokine CCL5; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Neoplasm Transplantation; Pain; Pain Measurement; Phenanthrenes; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; Spinal Cord

Mitochondrial disorders are highly heterogeneous conditions characterized by defects of the mitochondrial respiratory chain. Pharmacological activation of mitochondrial biogenesis has been proposed as an effective means to correct the biochemical defects and ameliorate the clinical phenotype in these severely disabling, often fatal, disorders. Pathways related to mitochondrial biogenesis are targets of Sirtuin1, a NAD(+)-dependent protein deacetylase. As NAD(+) boosts the activity of Sirtuin1 and other sirtuins, intracellular levels of NAD(+) play a key role in the homeostatic control of mitochondrial function by the metabolic status of the cell. We show here that supplementation with nicotinamide riboside, a natural NAD(+) precursor, or reduction of NAD(+) consumption by inhibiting the poly(ADP-ribose) polymerases, leads to marked improvement of the respiratory chain defect and exercise intolerance of the Sco2 knockout/knockin mouse, a mitochondrial disease model characterized by impaired cytochrome c oxidase biogenesis. This strategy is potentially translatable into therapy of mitochondrial disorders in humans.

Topics: Animals; Dietary Supplements; Disease Models, Animal; Electron Transport Complex IV; Energy Metabolism; Enzyme Activation; Gene Expression; Mice; Mice, Knockout; Mitochondria; Mitochondrial Diseases; Molecular Chaperones; NAD; Niacinamide; Oxidative Phosphorylation; Phenanthrenes; Phenotype; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Pyridinium Compounds; Sirtuin 1

To investigate the effects of cryptotanshinone (CRY), an active component of Chinese medicine, on ovarian androgen production, insulin resistance (IR), and glucose metabolism in mice.. Animal model and in vitro tissue model.. University-affiliated laboratory.. Mice.. Ovarian IR was induced by dexamethasone (DEX) in vivo. Animals were randomized to receive CRY treatment for 3 days or not. Ovulation rates, serum steroid levels, and glucose uptake in ovaries were quantified, and proteins in the phosphatidylinositol 3-hydroxy kinase pathway were measured. In vitro ovarian IR was also induced by DEX for 3 days. Ovarian steroid hormone secretion and glucose uptake were measured, and the hormone-synthesizing enzymes were determined by semiquantitative reverse transcription-polymerase chain reaction.. Ovarian glucose uptake, in vivo ovulation rate, serum and culture medium steroid level, and molecular expression of phosphatidylinositol 3-hydroxy kinase and steroidogenic enzymes.. Dexamethasone significantly increased ovulation rates in vivo and increased T and E2 production and decreased ovarian glucose uptake in vivo and in vitro. Cryptotanshinone significantly reduced ovulation rates in vivo and decreased T and estrogen production in vitro. Cryptotanshinone attenuated the inhibition of DEX on AKT2 and suppressed the up-regulation of CYP11 and CYP17 expression by DEX.. Cryptotanshinone reversed DEX-induced androgen excess and ovarian IR in mice through activation of insulin signaling and the regulation of glucose transporters and hormone-synthesizing enzymes. This suggests a potential role for CRY in treating the ovulatory dysfunction associated with PCOS.

Topics: Animals; Blood Glucose; Dexamethasone; Disease Models, Animal; Estradiol; Female; Gene Expression Regulation, Enzymologic; Glucose Metabolism Disorders; Glucose Transport Proteins, Facilitative; Insulin; Insulin Resistance; Mice; Ovary; Ovulation; Phenanthrenes; Phosphatidylinositol 3-Kinase; Polycystic Ovary Syndrome; Proto-Oncogene Proteins c-akt; Signal Transduction; Steroid 17-alpha-Hydroxylase; Testosterone; Tissue Culture Techniques

Triptolide, a major bioactive ingredient of a widely used herbal medicine, has been shown to possess multiple pharmacological functions, including potential neuroprotective effects pertinent to Alzheimer's disease (AD) in vitro. However, the therapeutic potential of triptolide for AD in vivo has not been thoroughly evaluated. In the present study, we investigated the impact of peripherally administered triptolide on AD-related behavior and neuropathology in APPswe/PS1ΔE9 (APP/PS1) mice, an established model of AD. Our results showed that two-month treatment with triptolide rescued cognitive function in APP/PS1 mice. Immunohistochemical analyses indicated that triptolide treatment led to a significant decrease in amyloid-β (Aβ) deposition and neuroinflammation in treated mice. In contrast to previous findings in vitro, biochemical analyses showed that triptolide treatment did not significantly affect the production pathway of Aβ in vivo. Intriguingly, further analyses revealed that triptolide treatment upregulated the level of insulin-degrading enzyme, a major Aβ-degrading enzyme in the brain, indicating that triptolide treatment reduced Aβ pathology by enhancing the proteolytic degradation of Aβ. Our findings demonstrate that triptolide treatment ameliorates key behavioral and neuropathological changes found in AD, suggesting that triptolide may serve as a potential therapeutic agent for AD.

Topics: Alzheimer Disease; Amyloid beta-Protein Precursor; Animals; Anxiety; Brain; Cognition; Disease Models, Animal; Diterpenes; Epoxy Compounds; Habituation, Psychophysiologic; Inflammation; Memory Disorders; Mice; Mice, Transgenic; Phenanthrenes; Presenilin-1; Protein Processing, Post-Translational; Proteolysis; Up-Regulation

Dinaciclib is a potent CDK1, 2, 5 and 9 inhibitor being developed for the treatment of cancer. Additional understanding of antitumor mechanisms and identification of predictive biomarkers are important for its clinical development. Here we demonstrate that while dinaciclib can effectively block cell cycle progression, in vitro and in vivo studies, coupled with mouse and human pharmacokinetics, support a model whereby induction of apoptosis is a main mechanism of dinaciclib's antitumor effect and relevant to the clinical duration of exposure. This was further underscored by kinetics of dinaciclib-induced downregulation of the antiapoptotic BCL2 family member MCL1 and correlation of sensitivity with the MCL1-to-BCL-xL mRNA ratio or MCL1 amplification in solid tumor models in vitro and in vivo. This MCL1-dependent apoptotic mechanism was additionally supported by synergy with the BCL2, BCL-xL and BCL-w inhibitor navitoclax (ABT-263). These results provide the rationale for investigating MCL1 and BCL-xL as predictive biomarkers for dinaciclib antitumor response and testing combinations with BCL2 family member inhibitors.

Topics: Aniline Compounds; Animals; Antineoplastic Agents; Apoptosis; bcl-X Protein; Bridged Bicyclo Compounds, Heterocyclic; Cell Cycle Checkpoints; Cell Line, Tumor; Cyclic N-Oxides; Disease Models, Animal; Diterpenes; Drug Resistance, Neoplasm; Drug Synergism; Epoxy Compounds; Female; Gene Dosage; Humans; Indolizines; Male; Mice; Myeloid Cell Leukemia Sequence 1 Protein; Neoplasms; Phenanthrenes; Pyridinium Compounds; RNA, Messenger; Sulfonamides; Xenograft Model Antitumor Assays

The complex pathogenesis of Alzheimer's disease (AD) involves multiple contributing factors, including amyloid β (Aβ) peptide accumulation, inflammation and oxidative stress. Effective therapeutic strategies for AD are still urgently needed. Triptolide is the major active compound extracted from Tripterygium wilfordii Hook.f., a traditional Chinese medicinal herb that is commonly used to treat inflammatory diseases. The 5-month-old 5XFAD mice, which carry five familial AD mutations in the β-amyloid precursor protein (APP) and presenilin-1 (PS1) genes, were treated with triptolide for 8 weeks. We observed enhanced spatial learning performances, and attenuated Aβ production and deposition in the brain. Triptolide also inhibited the processing of amyloidogenic APP, as well as the expression of βAPP-cleaving enzyme-1 (BACE1) both in vivo and in vitro. In addition, triptolide exerted anti-inflammatory and anti-oxidative effects on the transgenic mouse brain. Triptolide therefore confers protection against the effects of AD in our mouse model and is emerging as a promising therapeutic candidate drug for AD.

Topics: Alzheimer Disease; Amyloid beta-Protein Precursor; Amyloid Precursor Protein Secretases; Animals; Anti-Inflammatory Agents; Antioxidants; Aspartic Acid Endopeptidases; Brain; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Humans; Immunosuppressive Agents; Learning; Maze Learning; Memory; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Phenanthrenes; Plant Extracts

The mechanism underlying small-for-size graft failure after reperfusion is still unknown. Toll-like receptor 4 (TLR4) has attracted a great deal of attention in inflammation and allograft rejection in recent years. Medicinally, triptolide has anti-inflammatory, immunosuppressive, and antineoplastic activities. In the present study, we studied the effect of triptolide on TLR4 expression in small-for-size grafts.. The potential inhibitory effect of triptolide on TLR4 messenger ribonucleic acid and protein in a mouse model of small-for-size liver graft injury was assessed. We also assessed the expression of tumor necrosis factor α and interleukin-6 in this model.. The expression of hepatic TLR4 messenger ribonucleic acid and protein and downstream mediators, such as tumor necrosis factor α and interleukin-6, were reduced in the triptolide pretreatment groups (50 and 100 μg) in small-for-size liver grafts. In these same triptolide pretreatment groups, edema and necrosis were reduced and the levels of alanine aminotransferase and aspartate aminotransferase were significantly decreased in these small-for-size liver grafts.. This study showed that triptolide might inhibit TLR4 in vivo and that pretreatment with triptolide could inhibit TLR4 activation and reduce small-for-size liver graft injury.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Blotting, Western; Disease Models, Animal; Diterpenes; Epoxy Compounds; Gene Expression Regulation; Graft Rejection; Immunosuppressive Agents; Liver; Liver Transplantation; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; RNA, Messenger; Toll-Like Receptor 4

Given the importance of inflammation in the onset and progression of diabetic cardiomyopathy, we investigated the potential protective effects of triptolide, an anti-inflammatory agent, in streptozotocin-induced diabetic rat model and in H9c2 rat cardiac cells exposed to high glucose.. Diabetic rats were treated with triptolide (100, 200, or 400 μg/kg/day respectively) for 6 weeks. At the end of this study, after cardiac function measurements were performed, rats were sacrificed and their hearts were harvested for further histologic and molecular biologic analysis. Enhanced activity and expression of nuclear factor-kappaB (NF-κB) p65 in diabetic hearts were associated with increased inflammatory response, as demonstrated by increased pro-inflammatory cytokines, cell adhesion molecules and invading inflammatory cells, as well as increased fibrosis, in line with impaired left ventricular function. Triptolide attenuated these morpho-functional alterations. Furthermore, triptolide (20 ng/ml) also attenuated high glucose-induced inflammation in H9c2 rat cardiac cells.. Our data demonstrate that anti-inflammatory effects of triptolide involving the NF-κB signaling pathway can improve left ventricular function under diabetic conditions, suggesting triptolide treatment might be beneficial in diabetic cardiomyopathy.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Diabetes Mellitus, Experimental; Diabetic Cardiomyopathies; Disease Models, Animal; Diterpenes; Epoxy Compounds; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley; Ventricular Function, Left

The fear that schistosomes will become resistant to praziquantel (PZQ) motivates the search for alternatives to treat schistosomiasis. The antimalarials quinine (QN) and halofantrine (HF) possess moderate antischistosomal properties. The major metabolic pathway of QN and HF is through cytochrome P450 (CYP) 3A4. Accordingly, this study investigates the effects of CYP3A4 inhibitor, ketoconazole (KTZ), on the antischistosomal potential of these quinolines against Schistosoma mansoni infection by evaluating parasitological, histopathological, and biochemical parameters. Mice were classified into 7 groups: uninfected untreated (I), infected untreated (II), infected treated orally with PZQ (1,000 mg/kg) (III), QN (400 mg/kg) (IV), KTZ (10 mg/kg)+QN as group IV (V), HF (400 mg/kg) (VI), and KTZ (as group V)+HF (as group VI) (VII). KTZ plus QN or HF produced more inhibition (P<0.05) in hepatic CYP450 (85.7% and 83.8%) and CYT b5 (75.5% and 73.5%) activities, respectively, than in groups treated with QN or HF alone. This was accompanied with more reduction in female (89.0% and 79.3%), total worms (81.4% and 70.3%), and eggs burden (hepatic; 83.8%, 66.0% and intestinal; 68%, 64.5%), respectively, and encountering the granulomatous reaction to parasite eggs trapped in the liver. QN and HF significantly (P<0.05) elevated malondialdehyde levels when used alone or with KTZ. Meanwhile, KTZ plus QN or HF restored serum levels of ALT, albumin, and reduced hepatic glutathione (KTZ+HF) to their control values. KTZ enhanced the therapeutic antischistosomal potential of QN and HF over each drug alone. Moreover, the effect of KTZ+QN was more evident than KTZ+HF.

Topics: Animals; Anthelmintics; Disease Models, Animal; Drug Synergism; Female; Humans; Intestines; Ketoconazole; Liver; Male; Mice; Parasite Load; Phenanthrenes; Quinine; Schistosoma mansoni; Schistosomiasis mansoni; Treatment Outcome

To investigate the changes in retinal microglia and retinal ganglion cell (RGC) survival after long-term administration of a Chinese herb extract, triptolide, in a DBA/2J mice. DBA/2J mice (n = 96) were administered triptolide (n = 48) 25 µg/kg or vehicle (n = 48) and were judged at 7, 9, 11 months of age. Long-term triptolide treatment tended to attenuate the anterior segment pathology in experimental group, though intraocular pressure was not significantly different between the two groups. In the experimental group, RGC survival was improved (7, 9, 11 months: p = 0.035, 0.004, 0.014), and microglia activation was suppressed based on a more ramified appearance (9, 11 months: p = 0.024, 0.013) and a lower total microglial cell count (7, 9, 11 months: p = 0.028, 0.025, 0.014). Double-immunofluorescence staining revealed TNF? localized to microglia, TNFR1 localized to the RGCs and nerve fiber layer. These findings indicate that long-term triptolide administration suppressed microglia activation and improved RGC survival in DBA/2J mice.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Survival; Disease Models, Animal; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Female; Follow-Up Studies; Glaucoma; Immunohistochemistry; Immunosuppressive Agents; Intraocular Pressure; Mice; Mice, Inbred DBA; Microglia; Phenanthrenes; Receptors, Tumor Necrosis Factor, Type I; Retinal Ganglion Cells; Treatment Outcome

Recombinant tissue plasminogen activator (rt-PA) is the only pharmacological treatment approved for thrombolysis in patients suffering from ischemic stroke, but its administration aggravates the risk of hemorrhagic transformations. Experimental data demonstrated that rt-PA increases the activity of poly(ADP-ribose)polymerase (PARP). The aim of the present study was to investigate whether PJ34, a potent (PARP) inhibitor, protects the blood-brain barrier components from rt-PA toxicity. In our mouse model of cerebral ischemia, administration of rt-PA (10 mg/kg, i.v.) 6h after ischemia aggravated the post-ischemic degradation of ZO-1, claudin-5 and VE-cadherin, increased the hemorrhagic transformations (assessed by brain hemoglobin content and magnetic resonance imaging). Furthermore, rt-PA also aggravated ischemia-induced functional deficits. Combining PJ34 with rt-PA preserved the expression of ZO-1, claudin-5 and VE-cadherin, reduced the hemorrhagic transformations and improved the sensorimotor performances. In vitro studies also demonstrated that PJ34 crosses the blood-brain barrier and may thus exert its protective effect by acting on endothelial and/or parenchymal cells. Thus, co-treatment with a PARP inhibitor seems to be a promising strategy to reduce rt-PA-induced vascular toxicity after stroke.

Topics: Animals; Blood-Brain Barrier; Brain; Brain Ischemia; Disease Models, Animal; Mice; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Stroke; Tissue Plasminogen Activator

Anti-angiogenesis targeting VEGFR2 has been considered as an important strategy for cancer therapy. Tylophorine is known to possess anti-inflammatory and antitumor activity, but its roles in tumor angiogenesis, the key step involved in tumor growth and metastasis, and the involved molecular mechanism is still unknown. Therefore, we examined its anti-angiogenic effects and mechanisms in vitro and in vivo.. We used tylophorine and analyzed its inhibitory effects on human umbilical vein endothelial cells (HUVEC) in vitro and Ehrlich ascites carcinoma (EAC) tumor in vivo.. Tylophorine significantly inhibited a series of VEGF-induced angiogenesis processes including proliferation, migration, and tube formation of endothelial cells. Besides, it directly inhibited VEGFR2 tyrosine kinase activity and its downstream signaling pathways including Akt, Erk and ROS in endothelial cells. Using HUVECs we demonstrated that tylophorine inhibited VEGF-stimulated inflammatory responses including IL-6, IL-8, TNF-α, IFN-γ, MMP-2 and NO secretion. Tylophorine significantly inhibited neovascularization in sponge implant angiogenesis assay and also inhibited tumor angiogenesis and tumor growth in vivo. Molecular docking simulation indicated that tylophorine could form hydrogen bonds and aromatic interactions within the ATP-binding region of the VEGFR2 kinase unit.. Tylophorine exerts anti-angiogenesis effects via VEGFR2 signaling pathway thus, may be a viable drug candidate in anti-angiogenesis and anti-cancer therapies.

Topics: Alkaloids; Angiogenesis Inhibitors; Animals; Antineoplastic Agents, Phytogenic; Cell Movement; Cell Proliferation; Cell Survival; Cytokines; Disease Models, Animal; Female; Human Umbilical Vein Endothelial Cells; Humans; Indolizines; Male; Matrix Metalloproteinase 2; Mice; Molecular Conformation; Molecular Docking Simulation; Neoplasms; Neovascularization, Physiologic; Nitric Oxide; Phenanthrenes; Protein Binding; Protein Interaction Domains and Motifs; Signal Transduction; Tumor Burden; Tylophora; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2; Xenograft Model Antitumor Assays

The transient receptor potential melastatin 4 (TRPM4) channel is expressed in the sinoatrial node, but its physiologic roles in this tissue with cardiac pacemaker properties remain unknown. This Ca(2+)-activated nonselective cation channel (NSCCa) induces cell depolarization at negative potentials. It is implicated in burst generation in neurons and participates in induction of ectopic beating in cardiac ventricular preparations submitted to hypoxia/reoxygenation. Accordingly, TRPM4 may participate in action potential (AP) triggering in the sinoatrial node.. The purpose of this study was to investigate the influence of TRPM4 on spontaneous heart beating.. Spontaneous APs were recorded using intracellular microelectrodes in mouse, rat, and rabbit isolated right atria.. In the spontaneously beating mouse atrium, superfusion of the TRPM4-specific inhibitor 9-phenanthrol produced a concentration-dependent reduction in AP rate (maximal reduction = 62% that of control; EC50 = 8 × 10(-6) mol●L(-1)) without affecting other AP parameters. These effects were absent in TRPM4(-/-) mice. 9-Phenanthrol exerted a rate-dependent reduction with a higher effect at low rates. Similar results were obtained in rat. Moreover, application of 9-phenanthrol produced a reduction in diastolic depolarization slope in rabbit sinus node pacemaker cells.. These data showed that TRPM4 modulates beating rate. Pacemaker activity in the sinoatrial node results from the slow diastolic depolarization slope due to the "funny" current, Na/Ca exchange, and a Ca(2+)-activated nonselective cation current, which can be attributable in part to TRPM4 that may act against bradycardia.

Topics: Animals; Bradycardia; Disease Models, Animal; Female; Gene Expression Regulation; Heart Atria; Heart Rate; Mice; Mice, Inbred C57BL; Patch-Clamp Techniques; Phenanthrenes; Protein Kinase Inhibitors; Rabbits; Rats; Sinoatrial Node; TRPM Cation Channels

Myocardial infarction (MI) is a cause of high morbidity and mortality in the world. Sodium tanshinone IIA sulphonate (STS) has been well used in Oriental medicine for treating cardiovascular diseases, however, the underlying mechanisms remain unclear. Alterations of circulating lipid profiles, increased fatty acid β-oxidation and oxidative stress play most important roles in the pathogenesis of MI. The present study aims to elucidate whether STS possesses cardioprotective effect against MI driven by isoproterenol (ISO), and to investigate its potential mechanisms of action.. MI was induced by subcutaneous injection of ISO (85 mg/kg at interval of 24 h for 2 consecutive days) to rats. The rats were randomly divided into 6 groups: (1) control; (2) ISO; (3) STS (16 mg/kg) +control; (4-6) STS (16, 8, 4 mg/kg) +ISO.. Our study showed that STS could ameliorate cardiac dysfunction and variation of myocardial zymogram, up-regulate antioxidant systems, and maintain the levels of circulating lipids driven by supramaximal doses ISO as well. Moreover, modulation of redox-sensitive extracellular signal-regulated kinase1/2 (ERK1/2)/Nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) and AMP-activated protein kinase (AMPK)/acetyl CoA carboxylase (ACC)/carnitine palmitoyltransferase (CPT) 1 pathways were involved in STS induced cardioprotection.. STS exerts strong favorable cardioprotective action. Additionally, the properties of STS, such as anti-dyslipidemia, anti-oxidant and inhibition of fatty acid β-oxidation, may be the mechanisms underlying the observed results.

Topics: Animals; Antioxidants; Cardiotonic Agents; Disease Models, Animal; Fatty Acids; Gene Expression Regulation; Heart; Hemodynamics; Isoproterenol; Lipids; Male; Myocardial Infarction; Myocardium; Oxidation-Reduction; Oxygen; Phenanthrenes; Plant Extracts; Rats; Rats, Sprague-Dawley; Time Factors

Regulatory T cells play a key role in suppressing tumor immunity. Triptolide, a major active component isolated from the Chinese medicinal herb Tripterygium wilfordii, has been proven to possess multiple antitumor activities. Here, we investigated the effect of triptolide on regulatory T cells and on the level of IL-10, transforming growth factor-β, and vascular endothelial growth factor in tumor-bearing mice. Fifty male C57BL/6 mice were randomly grouped as follows: normal control group, model group with B16-F10 cells implanted, and three treatment groups with cyclophosphamide, triptolide-high dose, triptolide-low dose. The proportion of regulatory T cells in the spleen and axillary lymph nodes was evaluated by flow cytometric analysis. Production of cytokines IL-10, transforming growth factor-β, and vascular endothelial growth factor in serum was measured using enzyme-labeled immunosorbent assay kits. The mRNA levels of Foxp3, IL-10, and transforming growth factor-β in the spleen and vascular endothelial growth factor in tumor tissue were detected by real-time PCR. The results showed that triptolide significantly decreased the proportion of regulatory T cells and lowered the Foxp3 level in the spleen and axillary lymph nodes of tumor-bearing mice. Production of IL-10 and transforming growth factor-β in peripheral blood, and the mRNA level of IL-10 and transforming growth factor-β in the spleen were also decreased. Additionally, triptolide could remarkably inhibit production of vascular endothelial growth factor in tumor-bearing mice. In conclusion, our study demonstrated that triptolide might inhibit tumor growth by inhibiting regulatory T cells and some cytokines such as IL-10 and transforming growth factor-β, as well as production of vascular endothelial growth factor.

Topics: Animals; Antineoplastic Agents, Phytogenic; Disease Models, Animal; Diterpenes; Down-Regulation; Drugs, Chinese Herbal; Epoxy Compounds; Forkhead Transcription Factors; Interleukin-10; Lymph Nodes; Male; Melanoma; Mice, Inbred C57BL; Phenanthrenes; Phytotherapy; RNA, Messenger; Spleen; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tripterygium; Vascular Endothelial Growth Factor A

To investigate the mechanisms underlying the protective effects of sodium tanshinone IIA sulfonate (STS) in an ischemia-reperfusion (I/R)-induced rat myocardial injury model.. Male SD rats were iv injected with STS, STS+LY294002 or saline (NS) for 15 d. Then the hearts were subjected to 30 min of global ischemia followed by 2 h of reperfusion. Cardiac function, infarction size and area at risk were assessed. Cell apoptosis was evaluated with TUNEL staining, DNA laddering and measuring caspase-3 activity. In addition, isolated cardiomyocytes of neonatal rats were pretreated with the above drugs, then exposed to H2O2 (200 mol/L) for 1 h. Cell apoptosis was detected using flow cytometric assay. The levels of p-Akt, p-FOXO3A and Bim were examined with immunoblotting.. Compared to NS group, administration of STS (20 mg/kg) significantly reduced myocardial infarct size (40.28%±5.36% in STS group vs 59.52%±7.28% in NS group), and improved the myocardial function as demonstrated by the increased values of dp/dtmax, LVDP and coronary flow at different reperfusion time stages. Furthermore, STS significantly decreased the rate of apoptotic cells (15.11%±3.71% in STS group vs 38.21%±7.83% in NS group), and reduced caspase-3 activity to nearly a quarter of that in NS group. Moreover, STS significantly increased the phosphorylation of Akt and its downstream target FOXO3A, and decreased the expression of pro-apoptotic gene Bim. Co-treatment with the PI3K inhibitor LY294002 (40 mg/kg) partially countered the protective effects induced by STS treatment. In isolated cardiomyocytes, STS exerted similar protective effects as shown in the ex vivo I/R model.. STS pretreatment reduces infarct size and improves cardiac function in an I/R-induced rat myocardial injury model via activation of Akt/FOXO3A/Bim-mediated signal pathway.

Topics: Animals; Animals, Newborn; Apoptosis; Apoptosis Regulatory Proteins; Bcl-2-Like Protein 11; Cardiotonic Agents; Chromones; Disease Models, Animal; Flow Cytometry; Forkhead Box Protein O3; Forkhead Transcription Factors; Gene Expression Regulation; In Situ Nick-End Labeling; Male; Membrane Proteins; Morpholines; Myocardial Infarction; Myocardial Reperfusion Injury; Myocytes, Cardiac; Phenanthrenes; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Signal Transduction

Tylophorine and analogs are phenanthroindolizidine alkaloids, several of which have been reported to have anticancer, antiviral, and anti-inflammatory properties. However, their function in the immune system remains widely unknown. Transcription factor Foxp3 is critical for the development and function of Treg, which down-regulates the immune system and maintains tolerance to self-antigens. In the present study, we defined a novel tylophorine analog, W-8, enhanced TGF-β-induced Foxp3 expression at the mRNA and the protein levels. Interestingly, W-8 synergistically increased the level of TGF-β-induced p-Smad3 through inhibition of the AKT/mTOR pathway and enhanced the demethylation of the promoter region of the Foxp3 through inhibition of the ERK pathway and DNMT1 expression. Moreover, administration of W-8 suppressed TNBS-induced murine colitis and increased Tregs in lymphoid tissues. Finally, W-8 enhanced conversion of naïve T cells to Tregs in vivo. In summary, our results defined a novel compound that enhanced Foxp3 expression through transcriptional and epigenetic programs, and it might serve as a therapeutic agent for inflammatory diseases.

Topics: Alkaloids; Animals; Blotting, Western; Colitis; Disease Models, Animal; Forkhead Transcription Factors; Indolizines; Lymphocyte Activation; Mice; Phenanthrenes; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; T-Lymphocytes, Regulatory; Up-Regulation

Microglia are the major immune cells in the central nervous system and play a key role in brain injury pathology. However, the role of activated microglia after subacute cerebral ischemia (SCI) remains unknown. To address this issue, we established a permanent middle cerebral artery occlusion (pMCAO) rat model and treated pMCAO rats with N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-N,N-dimethylacetamide (PJ34) (an inhibitor of microglial activation), or with vehicle alone. Finally, we determined the differences between the PJ34-and vehicle-treated rats with respect to neurological deficits, infarct volume, neuronal loss and the expression of CD11b (a marker of microglial activation), glial cell line-derived neurotrophic factor (GDNF) and tumor necrosis factor-α (TNF-α) at 1, 3 and 7 days after treatment. We found that the PJ34-treated rats had more severe neurological deficits and a larger infarct volume and exhibited a decreased CD11b expression, more neuronal loss, decreased expression of GDNF mRNA and protein but increased expression of TNF-α mRNA and protein compared with the vehicle-treated rats at 3 and 7 days after treatment. These results indicate that activated microglia provide a neuroprotective role through balancing GDNF and TNF-α expression following SCI.

Topics: Animals; Brain Ischemia; CD11b Antigen; Disease Models, Animal; Glial Cell Line-Derived Neurotrophic Factor; Infarction, Middle Cerebral Artery; Male; Microglia; Phenanthrenes; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha

Triptolide, isolated from the herb Tripterygium wilfordii, has been shown to potently induce apoptosis in various malignant cells by inhibiting RNA synthesis and nuclear factor-κB activity. Previously, we showed that triptolide promotes apoptosis in acute myeloid leukemia (AML) cells via the mitochondria-mediated pathway, in part, by decreasing levels of the anti-apoptotic proteins XIAP and Mcl-1. MRx102 is a triptolide derivative, currently in preclinical development. Here we show that MRx102 potently promoted apoptosis in AML cell lines, with EC(50) values of 14.5±0.6 nM and 37.0±0.9 nM at 48 h for OCI-AML3 and MV4-11 cells, respectively. MRx102, at low nanomolar concentrations, also induced apoptosis in bulk, CD34(+) progenitor, and more importantly, CD34(+)CD38(-) stem/progenitor cells from AML patients, even when they were protected by coculture with bone marrow derived mesenchymal stromal cells. MRx102 decreased XIAP and Mcl-1 protein levels and inhibited RNA synthesis in OCI-AML3 cells. In vivo, MRx102 greatly decreased leukemia burden and increased survival time in non-obese diabetic/severe combined immunodeficiency mice harboring Ba/F3-ITD cells. Collectively, we demonstrated that MRx102 has potent antileukemic activity both in vitro and in vivo, has the potential to eliminate AML stem/progenitor cells and overcome microenvironmental protection of leukemic cells, and warrants clinical investigation.

Topics: Animals; Antineoplastic Agents, Alkylating; Apoptosis; Bone Marrow; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Leukemia, Myeloid, Acute; Male; Mice; Mice, Inbred NOD; Mice, SCID; Myeloid Cell Leukemia Sequence 1 Protein; Neoplastic Stem Cells; Phenanthrenes; Proto-Oncogene Proteins c-bcl-2; Transcription, Genetic; Tumor Microenvironment; X-Linked Inhibitor of Apoptosis Protein; Xenograft Model Antitumor Assays

Spinal muscular atrophy (SMA) is a progressive neuromuscular disease. Since disease severity is related to the amount of survival motor neuron (SMN) protein, up-regulated functional SMN protein levels from the SMN2 gene are considered a major SMA drug-discovery strategy. In this study, we investigated the possible effects of triptolide, a diterpene triepoxide purified from Tripterygium wilfordii Hook. F., as a new compound for increasing SMN protein.. The effects and mechanisms of triptolide on the production of SMA protein were determined by cell-based assays using the motor neuronal cell line NSC34 and skin fibroblasts from SMA patients. Wild-type (Smn(+/+) SMN2(-/-) , C57BL/6) and SMA-like (Smn(-/-) SMN2) mice were injected with triptolide (0.01 or 0.1 mg·kg(-1) ·day(-1) , i.p.) and their survival rate and level of change in SMN protein in neurons and muscle tissue measured.. In NSC34 cells and human SMA fibroblasts, pM concentrations of triptolide significantly increased SMN protein expression and the levels of SMN complex component (Gemin2 and Gemin3). In human SMA fibroblasts, triptolide increased SMN-containing nuclear gems and the ratio of full-length transcripts (FL-SMN2) to SMN2 transcripts lacking exon 7 (SMN2Δ7). Furthermore, in SMA-like mice, triptolide significantly increased SMN protein levels in the brain, spinal cord and gastrocnemius muscle. Furthermore, triptolide treatment increased survival and reduced weight loss in SMA-like mice.. Triptolide enhanced SMN protein production by promoting SMN2 activation, exon 7 inclusion and increasing nuclear gems, and increased survival in SMA mice, which suggests triptolide might be a potential candidate for SMA therapy.

Topics: Animals; Apoptosis; Blotting, Western; Body Weight; Cell Culture Techniques; Cell Line; Cell Survival; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Epoxy Compounds; Fibroblasts; Gemini of Coiled Bodies; Humans; Kaplan-Meier Estimate; Mice; Mice, Knockout; Molecular Structure; Motor Neurons; Muscular Atrophy, Spinal; Neuroprotective Agents; Phenanthrenes; Real-Time Polymerase Chain Reaction; Survival of Motor Neuron 2 Protein; Transcription, Genetic; Tripterygium; Up-Regulation

Under conditions of increased oxidative stress, such as pre-eclampsia and diabetes, overstimulation of PARP leads to endothelial dysfunction. Inhibition of PARP has been demonstrated to reverse the vascular dysfunction associated with diabetes in vivo. The present study was carried out to investigate the role of PARP in mediating the endothelial dysfunction associated with pre-eclampsia.. Uteroplacental perfusion was surgically reduced in pregnant rats to produce the reduced uterine perfusion pressure (RUPP) rat model of pre-eclampsia and the PARP inhibitor, PJ34, was administered either before or after surgery. Mean arterial BP and vascular function were measured in normal pregnant (NP) and both control and PJ34-treated RUPP rats. Mesenteric vessels from NP rats were incubated with either 3% RUPP or NP plasma alone or in combination with PJ34. Finally, immunohistochemical staining was carried out to measure nitrotyrosine (byproduct of peroxynitrite) immunoreactivity.. RUPP rats were characterized by hypertension, fetal growth restriction and endothelial dysfunction when compared with NP rats. PJ34 administered in vivo before, but not after, surgery prevented the development of both endothelial dysfunction and hypertension. RUPP plasma-induced impaired vasorelaxation was prevented following co-incubation with PJ34 in vitro. Furthermore, the protective effect of PARP inhibition in vivo was accompanied by a reduction in nitrotyrosine immunoreactivity.. PJ34 prevented the development of both endothelial dysfunction and hypertension and reduced vascular nitrotyrosine immunoreactivity, thus suggesting a role for oxidative-nitrosative stress/PARP activation in the aberration in both vascular and haemodynamic function in this rat model of pre-eclampsia.

Topics: Animals; Disease Models, Animal; Endothelium, Vascular; Enzyme Inhibitors; Female; Hypertension; Mesenteric Arteries; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Pre-Eclampsia; Pregnancy; Rats; Rats, Sprague-Dawley

Traumatic brain injury (TBI) is characterized by neuroinflammation, brain edema, and cerebral damage leading to impairment of neurobehavioral function. Triptolide (PG-490), a diterpenoid component from Tripterygium wilfordii Hook F., has anti-inflammatory properties. Whether triptolide has neuroprotective functions when treating TBI is unclear. To investigate this possibility, Sprague-Dawley rats were treated with triptolide immediately after TBI had been induced by a controlled cortical impact procedure or after a sham procedure. TBI produced neuroinflammation when measured on day 1 after TBI, induced cerebral damage when measured on day 1 and day 3, and impaired neurobehavioral functioning over a 28-day observation period. Triptolide suppressed TBI-induced increases in contusion volume, cell apoptosis, edema and the levels of various pro-inflammatory mediators in the brain. Thriptolide reversed the TBI-induced decrease in brain levels of anti-inflammatory cytokine interleukin-10. Importantly, triptolide improved neurobehavioral outcomes regarding motor, sensory, reflex and balance function. We conclude that triptolide confers neuroprotection against TBI, at least in part, via its anti-inflammatory activity.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Brain Edema; Brain Injuries; Disease Models, Animal; Diterpenes; Epoxy Compounds; Inflammation Mediators; Male; Neurons; Neuroprotective Agents; Phenanthrenes; Rats; Rats, Sprague-Dawley; Recovery of Function

Pheochromocytomas (PHEOs) and paragangliomas (PGLs) are specific types of neuroendocrine tumors that originate in the adrenal medulla or sympathetic/parasympathetic paraganglia, respectively. Although these tumors are intensively studied, a very effective treatment for metastatic PHEO or PGL has not yet been established. Preclinical evaluations of novel therapies for these tumors are very much required. Therefore, in this study we tested the effect of triptolide (TTL), a potent nuclear factor-kappaB (NF-κB) inhibitor, on the cell membrane norepinephrine transporter (NET) system, considered to be the gatekeeper for the radiotherapeutic agent 131I-metaiodobenzylguanidine (131I-MIBG). We measured changes in the mRNA and protein levels of NET and correlated them with proapoptotic factors and metastasis inhibition. The study was performed on three different stable PHEO cell lines. We found that blocking NF-κB with TTL or capsaicin increased both NET mRNA and protein levels. Involvement of NF-κB in the upregulation of NET was verified by mRNA silencing of this site and also by using NF-κB antipeptide. Moreover, in vivo treatment with TTL significantly reduced metastatic burden in an animal model of metastatic PHEO. The present study for the first time shows how NF-κB inhibitors could be successfully used in the treatment of metastatic PHEO/PGL by a significant upregulation of NET to increase the efficacy of 131I-MIBG and by the induction of apoptosis.

Topics: Animals; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Gene Expression Regulation, Neoplastic; Humans; Mice; Mice, Nude; Neoplasm Metastasis; NF-kappa B; Norepinephrine Plasma Membrane Transport Proteins; Paraganglioma; Phenanthrenes; Pheochromocytoma; Rats; RNA Interference; Transcription, Genetic; Tumor Burden

Apoptosis, necrosis, and inflammation are hallmarks of cisplatin nephrotoxicity; however, the role and mechanisms of necrosis and inflammation remains undefined. As poly(ADP-ribose) polymerase 1 (PARP1) inhibition or its gene deletion is renoprotective in several renal disease models, we tested whether its activation may be involved in cisplatin nephrotoxicity. Parp1 deficiency was found to reduce cisplatin-induced kidney dysfunction, oxidative stress, and tubular necrosis, but not apoptosis. Moreover, neutrophil infiltration, activation of nuclear factor-κB, c-Jun N-terminal kinases, p38 mitogen-activated protein kinase, and upregulation of proinflammatory genes were all abrogated by Parp1 deficiency. Using proximal tubule epithelial cells isolated from Parp1-deficient and wild-type mice and pharmacological inhibitors, we found evidence for a PARP1/Toll-like receptor 4/p38/tumor necrosis factor-α axis following cisplatin injury. Furthermore, pharmacological inhibition of PARP1 protected against cisplatin-induced kidney structural/functional damage and inflammation. Thus, our findings suggest that PARP1 activation is a primary signal and its inhibition/loss protects against cisplatin-induced nephrotoxicity. Targeting PARP1 may offer a potential therapeutic strategy for cisplatin nephrotoxicity.

Topics: Acute Kidney Injury; Animals; Antineoplastic Agents; Apoptosis; Cells, Cultured; Cisplatin; Disease Models, Animal; Enzyme Activation; Enzyme Inhibitors; Gene Expression Regulation; Inflammation Mediators; JNK Mitogen-Activated Protein Kinases; Kidney Tubules; Male; Mice; Mice, Knockout; Necrosis; Nephritis; Neutrophil Infiltration; NF-kappa B; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Signal Transduction; Time Factors; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Due to the immunoinflammatory pathology of Alzheimer's disease (AD) brain, recent studies have begun to focus attention on the role of anti-inflammatory drugs or immunomodulators in AD. Triptolide isolated from the herb Tripterygium wilfordii Hook F has anti-inflammatory and immunosuppressive activities. In this study, we observed the effects of triptolide on synaptophysin expression in AD cellular model. AD cellular model was established by action of Aβ-stimulated microglial conditioned medium (MCM) on cultured rat hippocampal neurons (HN). Immunocytochemical staining, western blot and RT-PCR were used to observe the effects of triptolide at different dosages on the synaptophysin expression of hippocampal neurons in AD cellular model at different time points during incubation of cultures. After 24 h of cultivation, the expression level of synaptophysin in MCM/HN model group was decreased as compared with normal HN group and MCM/HN control group, and the expression level of synaptophysin in MCM/HN low-dose triptolide group and MCM/HN high-dose triptolide group was increased as compared with MCM/HN model group. It is concluded that triptolide can promote the synaptophysin expression of hippocampal neurons in the AD cellular model.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Animals; Anti-Inflammatory Agents, Non-Steroidal; Base Sequence; Cells, Cultured; Culture Media, Conditioned; Disease Models, Animal; Diterpenes; Epoxy Compounds; Gene Expression; Hippocampus; Immunologic Factors; Microglia; Neurons; Neuroprotective Agents; Peptide Fragments; Phenanthrenes; Protein Multimerization; Rats; RNA, Messenger; Synaptophysin; Vesicular Transport Proteins

Although TRAIL (tumor necrosis factor (TNF)-related apoptosis inducing ligand) is a well-known apoptosis inducer, we have previously demonstrated that acidic extracellular pH (pHe) switches TRAIL-induced apoptosis to regulated necrosis (or necroptosis) in human HT29 colon and HepG2 liver cancer cells. Here, we investigated the role of RIPK1 (receptor interacting protein kinase 1), RIPK3 and PARP-1 (poly (ADP-ribose) polymerase-1) in TRAIL-induced necroptosis in vitro and in concanavalin A (Con A)-induced murine hepatitis. Pretreatment of HT29 or HepG2 with pharmacological inhibitors of RIPK1 or PARP-1 (Nec-1 or PJ-34, respectively), or transient transfection with siRNAs against RIPK1 or RIPK3, inhibited both TRAIL-induced necroptosis and PARP-1-dependent intracellular ATP depletion demonstrating that RIPK1 and RIPK3 were involved upstream of PARP-1 activation and ATP depletion. In the mouse model of Con A-induced hepatitis, where death of mouse hepatocytes is dependent on TRAIL and NKT (Natural Killer T) cells, PARP-1 activity was positively correlated with liver injury and hepatitis was prevented both by Nec-1 or PJ-34. These data provide new insights into TRAIL-induced necroptosis with PARP-1 being active effector downstream of RIPK1/RIPK3 initiators and suggest that pharmacological inhibitors of RIPKs and PARP-1 could be new treatment options for immune-mediated hepatitis.

Topics: Adenosine Triphosphate; Animals; Apoptosis; Cell Line; Disease Models, Animal; Hep G2 Cells; Hepatocytes; HT29 Cells; Humans; Hydrogen-Ion Concentration; Imidazoles; Indoles; Killer Cells, Natural; Mice; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Reactive Oxygen Species; Receptor-Interacting Protein Serine-Threonine Kinases; RNA Interference; RNA, Small Interfering; TNF-Related Apoptosis-Inducing Ligand

Peripheral nerve injury generally results in spinal neuronal and glial plastic changes associated with chronic behavioral hypersensitivity. Spinal mitogen-activated protein kinases (MAPKs), eg, p38 or extracellular signal-regulated kinases (ERKs), are instrumental in the development of chronic allodynia in rodents, and new p38 inhibitors have shown potential in acute and neuropathic pain patients. We have previously shown that the cannabinoid type 2 receptor agonist JWH015 inhibits ERK activity by inducing MAPK phosphatase (MKP)-1 and MKP-3 (the major regulators of MAPKs) in vitro in microglial cells. Therefore, we decided to investigate the role of these phosphatases in the mechanisms of action of JWH015 in vivo using the rat L5 nerve transection model of neuropathic pain. We observed that peripheral nerve injury reduced spinal MKP-1/3 expression and activity and that intrathecal JWH015 reduced established L5 nerve injury-induced allodynia, enhanced spinal MKP-1/3 expression and activity, and reduced the phosphorylated form of p38 and ERK-1/2. Triptolide, a pharmacological blocker of MKP-1 and MKP-3 expression, inhibited JWH015's effects, suggesting that JWH015 exerts its antinociceptive effects by modulating MKP-1 and MKP-3. JWH015-induced antinociception and MKP-1 and MKP-3 expression were inhibited by the cannabinoid type 2 receptor antagonist AM630. Our data suggest that MKP-1 and MKP-3 are potential targets for novel analgesic drugs.. MAPKs are pivotal in the development of chronic allodynia in rodent models of neuropathic pain. A cannabinoid type 2 receptor agonist, JWH015, reduced neuropathic allodynia in rats by reducing MAPK phosphorylation and inducing spinal MAPK phosphatases 1 and 3, the major regulators of MAPKs.

Topics: 4-Nitrophenylphosphatase; Animals; Disease Models, Animal; Diterpenes; Dual Specificity Phosphatase 1; Dual Specificity Phosphatase 6; Epoxy Compounds; Gene Expression Regulation; Hyperalgesia; Immunosuppressive Agents; Indoles; Male; Mitogen-Activated Protein Kinase Phosphatases; Nerve Tissue Proteins; Neuralgia; Phenanthrenes; Rats; Rats, Sprague-Dawley; Receptor, Cannabinoid, CB2; Signal Transduction; Spinal Cord; Time Factors

To investigate the neuroprotective effects of LLDT-67, a novel derivative of triptolide, in MPTP-induced mouse Parkinson's disease (PD) models and in primary cultured astrocytes, and to elucidate the mechanisms of the action.. In order to induce PD, C57BL/6 mice were injected MPTP (30 mg/kg, ip) daily from d 2 to d 6. MPTP-induced behavioral changes in the mice were examined using pole test, swimming test and open field test. The mice were administered LLDT-67 (1, 2, or 4 mg/kg, po) daily from d 1 to d 11. On d 12, the mice were decapitated and brains were collected for immunohistochemistry study and measuring monoamine levels in the striatum. Primary cultured astrocytes from the cortices of neonatal C57BL/6 mouse pups were prepared for in vitro study.. In MPTP-treated mice, administration of LLDT-67 significantly reduced the loss of tyrosine hydroxylase-positive neurons in the substantia nigra, and ameliorated the behavioral changes. LLDT-67 (4 mg/kg) significantly increased the expression of NGF in astrocytes in the substantia nigra and striatum of the mice. Furthermore, administration of LLDT-67 caused approximately 2-fold increases in the phosphorylation of TrkA at tyrosine 751, and marked increases in the phosphorylation of AKT at serine 473 as compared with the mice model group. In the cultured astrocytes, LLDT-67 (1 and 10 nmol/L) increased the NGF levels in the culture medium by 179% and 160%, respectively.. The neuroprotective effect of LLDT-67 can be mostly attributed to its ability to enhance NGF synthesis in astrocytes in the midbrain and to rescue dopaminergic neurons indirectly through TrkA activation.

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Animals, Newborn; Astrocytes; Cells, Cultured; Corpus Striatum; Disease Models, Animal; Diterpenes; Dopaminergic Neurons; Dose-Response Relationship, Drug; Male; Mice; Mice, Inbred C57BL; Nerve Growth Factor; Neuroprotective Agents; Parkinsonian Disorders; Phenanthrenes; Phosphorylation; Proto-Oncogene Proteins c-akt; Receptor, trkA; Substantia Nigra; Time Factors; Up-Regulation

Poly(ADP-ribose) polymerase (PARP) is well known to be an enzyme that repairs damaged DNA and also induces cell death when overactivated. It has been reported that PARP plays a significant role in burn and smoke inhalation injury, and the pathophysiology is thought to be localized in the airway during early stages of activation. Therefore, we hypothesized that local inhibition of PARP in the airway by direct delivery of low dose PJ-34 [poly(ADP-ribose) polymerase inhibitor] into the bronchial artery would attenuate burn and smoke-induced acute lung injury. The bronchial artery in sheep was cannulated in preparation for surgery. After a 5-7 day recovery period, sheep were administered a burn and inhalation injury. Adult female sheep (n=19) were divided into four groups following the injury: (1) PJ-34 group A: 1h post-injury, PJ-34 (0.003mg/kg/h, 2mL/h) was continuously injected into the bronchial artery, n=5; (2) PJ-34 group B: 1h post-injury, PJ-34 (0.03mg/kg/h, 2mL/h) was continuously injected into bronchial artery, n=4; (3) CONTROL GROUP: 1h post-injury, an equivalent amount of saline was injected into the bronchial artery, n=5; (4) Sham group: no injury, no treatment, same operation and anesthesia, n=5. After injury, all animals were placed on a ventilator and fluid resuscitated equally. Pulmonary function as evaluated by measurement of blood gas analysis, pulmonary mechanics, and pulmonary transvascular fluid flux was severely deteriorated in the control group. However, the above changes were markedly attenuated by PJ-34 infusion into the bronchial artery (P/F ratio at 24h: PJ-34 group A 398±40*, PJ-34 group B 438±41*†‡, Control 365±58*, Sham 547±47; * vs. sham [p<0.05], † vs. control [p<0.05], ‡ vs. PJ-34 group A [p<0.05]). Our data strongly suggest that local airway production of poly(ADP-ribose) polymerase contributes to pulmonary dysfunction following smoke inhalation and burn.

Topics: Acute Lung Injury; Animals; Bronchial Arteries; Burns; Disease Models, Animal; Enzyme Inhibitors; Female; Lung; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Respiratory Function Tests; Sheep; Smoke Inhalation Injury

Airway remodelling contributes to increased morbidity and mortality in asthma. We have reported that triptolide, the major component responsible for the immunosuppressive and anti-inflammatory effects of Tripterygium wilfordii Hook F, inhibited pulmonary inflammation in patients with steroid-resistant asthma. In the present study, we investigated whether triptolide inhibits airway remodelling in a mouse asthma model and observed the effects of triptolide on the transforming growth factor-β₁ (TGF-β₁)/Smad pathway in ovalbumin (OVA)-sensitized mice. BALB/c mice were sensitized to intraperitoneal OVA followed by repetitive OVA challenge for 8 weeks. Treatments included triptolide (40 μg/kg) and dexamethasone (2 mg/kg). The area of bronchial airway (WAt/basement membrane perimeter) and smooth muscle (WAm/basement membrane perimeter), mucus index and collagen area were assessed 24 hr after the final OVA challenge. Levels of TGF-β(1) were assessed by immunohistology and ELISA, levels of TGF-β(1) mRNA were measured by RT-PCR, and levels of pSmad2/3 and Smad7 were assessed by Western blot. Triptolide and dexamethasone significantly reduced allergen-induced increases in the thickness of bronchial airway and smooth muscle, mucous gland hypertrophy, goblet cell hyperplasia and collagen deposition. Levels of lung TGF-β(1) , TGF-β(1) mRNA and pSmad2/3 were significantly reduced in mice treated with triptolide and dexamethasone, and this was associated with a significant increase in levels of Smad7. Triptolide may function as an inhibitor of asthma airway remodelling. It may be a potential drug for the treatment of patients with a severe asthma airway.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Blotting, Western; Bronchoalveolar Lavage Fluid; Dexamethasone; Disease Models, Animal; Diterpenes; Enzyme-Linked Immunosorbent Assay; Epoxy Compounds; Female; Immunosuppressive Agents; Mice; Mice, Inbred BALB C; Ovalbumin; Phenanthrenes; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Smad Proteins; Transforming Growth Factor beta1

Endothelium integrity is important for the normal functioning of vessels, the disruption of which can lead to disease. The blood-retinal barrier required for normal retinal function is compromised in diabetic retinopathy, causing retinal vascular leakage. Previously, we demonstrated the ability of Sac-0601[((2R,3S)-3-acetoxy-6-((3S,10R,13R,17R)-10,13-dimethyl-17-((R)-6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yloxy)-3,6-dihydro-2H-pyran-2-yl)methyl acetate], a pseudo-sugar derivative of cholesterol, to increase survival of retinal endothelial cells. In the present study, we evaluated the ability of Sac-0601 to prevent retinal vascular leakages in vitro and in vivo. Sac-0601 treatment blocked VEGF-induced formation of actin stress fibers and stabilized the cortical actin ring in retinal endothelial cells. It also inhibited degradation of occludin, an important tight junction protein, and blocked VEGF-induced disruption of its linear pattern at the cell border. The [(14)C] sucrose permeability assay demonstrated that Sac-0601 was able to prevent VEGF-induced retinal endothelial permeability. The compound inhibited the vascular leakage in retina of mice intravitreally injected with VEGF. And it also significantly reduced the leakage in retina of diabetic retinopathy mice model. Taken together, our findings suggest the potential therapeutic usefulness of Sac-0601 for retinal vascular permeability diseases.

Topics: Animals; Capillary Permeability; Cell Line; Diabetic Retinopathy; Disease Models, Animal; Endothelial Cells; Humans; Membrane Proteins; Mice; Occludin; Phenanthrenes; Pyrans; Retinal Vessels; Stress Fibers; Tight Junctions; Vascular Diseases; Vascular Endothelial Growth Factor A

Microsomal prostaglandin E synthase-1 (mPGES1) is critical for prostaglandin E(2) formation in ductus arteriosus (DA) and, accordingly, in its patency. We previously reported that mPGES1 deletion, unlike cyclo-oxygenase (COX) suppression, is not followed by upregulation of relaxant nitric oxide (NO). Consequently, we proposed that a mPGES1 inhibitor may be better than currently used COX inhibitors in managing premature infants with persistent DA (PDA).. To assess the effect of the mPGES1 inhibitor, 2-(6-chloro-1H-phenanthro[9,10d]imidazole-2-yl)isophthalonitrile (MF63) on DA ex vivo and in vivo (p.o. to the mother).. Experiments were carried out with mice bearing human mPGES1. We utilized isolated, wire-mounted DA for isometric recording and a whole-body freezing technique to assess the DA caliber as it occurs in vivo.. MF63 (10 μM) contracted the isolated DA. DA constriction was also seen in vivo after a single 10-mg kg(-1) dose. Conversely, a 30-mg kg(-1) dose gave inconsistent results, combining constriction with no effect. DA dilatation followed instead a repeated lower dose (twice daily for 3 days), and postnatal closure of the vessel was also delayed. Chronic pretreatment had no effect on endothelial NO synthase mRNA expression in fetal DA, nor did it modify the contraction to NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (100 μM).. MF63 has a dual action on DA, the constriction being associated with accessory dilatation. The latter effect should be explained before considering further a mPGES1 inhibitor for management of PDA.

Topics: Animals; Disease Models, Animal; Dose-Response Relationship, Drug; Ductus Arteriosus; Ductus Arteriosus, Patent; Enzyme Inhibitors; Female; Gene Knock-In Techniques; Humans; Imidazoles; Intramolecular Oxidoreductases; Mice; Mice, Inbred C57BL; Nitric Oxide Synthase Type III; Phenanthrenes; Pregnancy; Prostaglandin-E Synthases; Vasoconstriction; Vasodilation

The study is to test the efficacy of triptolide-coated stent on anti-inflammatory and inhibiting intimal hyperplasia in preventing restenosis.. A total of 15 triptolide-coated stents (TCSs), 12 drug-eluting stents (DESs), and 12 bare metal stents (BMSs) were deployed in porcine coronary arteries. Coronary angiography, histopathologic and inflammatory markers levels were analyzed at 4 week after stenting.. At 4-week follow-up, quantitative coronary angiography (QCA) showed that minimum lumen diameter was greater, and percentage stenosis and late lumen loss in TCS group and DES group were less than that of BMS group. Compared to BMS group, injury and inflammation score in TCS group and DES group were decreased, neointimal area significantly reduced and lumen area enlarged in TCS, DES group as well.. Triptolide-coated stent showed the effect of preventing restenosis via inhibition of neointimal hyperplasia and anti-inflammation acts after coronary angioplasty.

Topics: Analysis of Variance; Angioplasty, Balloon, Coronary; Animals; Biomarkers; Disease Models, Animal; Diterpenes; Drug-Eluting Stents; Epoxy Compounds; Immunosuppressive Agents; Phenanthrenes; Swine; Tunica Intima

Ischemia and reperfusion have been identified as a complex cascade of inflammatory mediators that are involved in the pathogenesis of hepatic injury. Triptolide (diterpenoid triepoxide), was extracted from a purified component of a traditional Chinese Medicine, Tripterygium wilfondii Hook F. Currently, triptolide has been shown to have anti-inflammatory, immunosuppressive, and antineoplastic activity. Accumulated data have shown that Th17 cells might contribute to the pathogenesis of liver diseases. Triptolide has been shown to reduce interleukin (IL)-17 expression in inflammatory bowel disease and arthritis. However, the role of triptolide in liver ischemia/reperfusion (I/R) and whether it can attenuate injury and the potential mechanism have not been investigated. Mice were treated with triptolide (0.1mg/kg) for 1 week or IL-17 antibody (50 μg/mouse) 2 days before ischemic insult. Partial warm ischemia was produced in the hepatic lobes of C57BL/6 mice for 90 min, followed by various periods of reperfusion. We demonstrated that IL-17 was involved in the inflammatory response to hepatic I/R injury, and that triptolide inhibited IL-17 generation and suppressed neutrophil migration after liver I/R injury through downregulation of signal transducer and activator of transcription 3 (STAT3) transcription. Also, triptolide pretreatment protected the liver from warm I/R injury, at least in part, mediated by the upregulation of Foxp3 expression. These results could pave the way for the use of triptolide as a novel agent to attenuate I/R injury.

Topics: Animals; Antibodies, Monoclonal; Cell Movement; Disease Models, Animal; Diterpenes; Epoxy Compounds; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Interleukin-17; Liver; Liver Diseases; Mice; Mice, Inbred C57BL; Neutrophils; Phenanthrenes; Phytotherapy; Reperfusion Injury; STAT3 Transcription Factor; Tripterygium

Current study was conducted to identify constituents of Taxus wallichiana Zucc. that might be responsible for its folk use in anti-inflammatory conditions. Taxusabietane A was isolated from the bark extract of Taxus wallichiana Zucc. Taxusabietane A was analyzed for in-vitro and in-vivo anti-inflammatory activities using Lipoxygenase (LOX) inhibition assay and carrageenan-induced paw edema model. Taxusabietane A revealed considerable LOX inhibitory activity with the IC(50) value being 57 ± 0.31. Standard compound Baicalein showed the IC(50) value being 22.1 ± 0.03 μM. Taxusabietane A also showed significant (5 and 10 mg/kg) anti-inflammatory activity induced by carrageenan. However, this study highlighted the potential of Taxusabietane A to be further explored as a new lead compound for management of conditions associated with inflammation.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Disease Models, Animal; Edema; Female; Flavanones; Lipoxygenase Inhibitors; Male; Phenanthrenes; Phytotherapy; Plant Bark; Plant Extracts; Rats; Rats, Wistar; Taxus

The root of Salvia miltiorrhiza Bunge, a well-known traditional Chinese medicine, has been used effectively for the treatment of cardiovascular diseases for a long time. The mechanisms underlying this therapeutic effect are not, however, fully understood. Tanshinone IIA (Tan IIA) is one of the major active components of this Chinese medicine. Therefore, the present study was performed to investigate whether Tan IIA, which has shown a cardio-protective capacity in myocardial ischemia, has an inhibitory effect on the inflammatory responses following myocardial infarction (MI) and its potential mechanisms. In an in vivo study, rat MI model was induced by permanent left anterior descending coronary artery (LAD) ligation. After the operation rats were divided into three groups (sham, MI and Tan IIA). Tan IIA was administered intragastrically at a dose of 60mg/kg body wt./day. One week later, rats were sacrificed and the hemodynamic, pathological and molecular biological indices were examined. In an in vitro study, the inflammatory model was established by TNF-alpha stimuli on cardiacmyocyte and cardiac fibroblasts. Tan IIA attenuates the MI pathological changes and improves heart function, and reduces expression of MCP-1, TGF-beta(1) and macrophage infiltration. Furthermore, Tan IIA could also decrease the expression of TNF-alpha and activation of nuclear transcription factor-kappa B (NF-kappaB). In vitro, Tan IIA could reduce MCP-1 and TGF-beta(1)secretion of cardiac fibroblasts. The present study demonstrated that the cardioprotective effects of Tan IIA might be attributed to its capacity for inhibiting inflammatory responses.

Topics: Abietanes; Animals; Anti-Inflammatory Agents; Chemokine CCL2; Coronary Vessels; Disease Models, Animal; Drugs, Chinese Herbal; Fibroblasts; Heart; Inflammation; Macrophages; Male; Myocardial Infarction; Myocardium; Myocytes, Cardiac; NF-kappa B; Phenanthrenes; Phytotherapy; Plant Roots; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

The aim of this work was to evaluate the suitability of ethosomes as carriers for the topical application of triptolide in a rat model of erythema.. We determined the optimal conditions for preparing ethosomes, and we measured their vesicle size by a laser particle-size analyzer and the efficiency of entrapment of triptolide by ultracentrifugation.. The in vitro percutaneous permeation of triptolide-loaded ethosomes was investigated by measuring diffusion across a sample of rat skin. To explore the transdermal delivery in vivo, we used a model in which erythema was induced in rats by methyl nicotinate and determined the change in erythema index caused by the anti-inflammatory activity of triptolide by a reflection spectrophotometer.. The optimal conditions for preparing triptolide ethosomes consisted of ultrasonication of 45% (v/v) ethanol and 2% (w/v) DPPC for 5 minutes, which produced an average vesicle size of 51.4 nm and an entrapment efficiency of 98%. This ethosomal formulation of triptolide caused the greatest in vitro 24-hour accumulation of triptolide (83.7%) with no permeation time delay, and it reduced erythema in vivo more rapidly and more completely than other formulations.. Ethosomes might be a promising carrier that would enable the beneficial properties of triptolide to be safely delivered in a topical formulation.

Topics: Administration, Cutaneous; Animals; Anti-Inflammatory Agents; Diffusion; Disease Models, Animal; Diterpenes; Drug Carriers; Drug Delivery Systems; Epoxy Compounds; Erythema; Ethanol; Female; Liposomes; Male; Particle Size; Permeability; Phenanthrenes; Rats; Rats, Sprague-Dawley

Several experiments were designed to determine whether the systemic, postischemic administration of PJ34,which is a poly-adenosine diphosphate (ADP)-ribose polymerase inhibitor, decreased tissue injury and inflammation after hind-limb ischemia reperfusion (I/R).. C57BL6 mouse limbs were subjected to 1.5 h ischemia followed by 24-h reperfusion. The treatment group (PJ) received intraperitoneal PJ34 (30 mg/kg) immediately before reperfusion, as well as 15 min and 2 h into reperfusion. The control group (CG) received lactated Ringer's alone at the same time intervals as PJ34 administration. The skeletal muscle levels of adenosine triphosphate (ATP), macrophage inflammatory protein-2 (MIP-2), keratinocyte derived chemokine (KC), and myeloperoxidase (MPO) were measured. Quantitative measurement of skeletal muscle tissue injury was assessed by microscopic analysis of fiber injury.. ATP levels were higher in limbs of PJ versus CG mice (absolute ATP: 4.7 +/- 0.35 vs 2.3 +/- 0.15-ng/mg tissue, P = .002). The levels of MIP-2, KC, and MPO were lower in PJ versus CG mice (MIP-2: 1.4 +/- 0.34 vs 3.67 +/- 0.67-pg/mg protein, P = .014; KC: 4.97 +/- 0.97 vs 12.65 +/- 3.05-pg/mg protein, P = .037; MPO: 46.27 +/- 10.53 vs 107.34 +/- 13.58-ng/mg protein, P = .008). Muscle fiber injury was markedly reduced in PJ versus CG mice (4.25 +/- 1.9% vs 22.68 +/- 3.0% total fibers, P = .0004).. Systemic postischemic administration of PJ34 preserved skeletal muscle energy levels, decreased inflammatory markers, and preserved tissue viability post-I/R. These results support PARP inhibition as a viable treatment for skeletal muscle I/R in a clinically relevant post hoc scenario.

Topics: Adenosine Triphosphate; Animals; Chemokine CXCL1; Chemokine CXCL2; Disease Models, Animal; Hindlimb; Ischemia; Male; Mice; Mice, Inbred C57BL; Muscle, Skeletal; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Reperfusion Injury

Autosomal dominant polycystic kidney disease (ADPKD), a major cause of end-stage renal failure, results from genetic mutation of either polycystin-1 (Pkd1) or polycystin-2 (Pkd2). In order to develop novel therapies to treat the advancement of disease progression, numerous rodent models of different genetic backgrounds are available to study cyst development.. Here, a Pkd1-floxed inducible mouse model using the interferon responsive Mx1Cre-recombinase was utilized to test the effect of the small molecule triptolide. Relative to other Pkd1 inactivation models, cyst progression in this neonatal to adult transition model is attenuated. Following the characterization of inducible cyst formation in these mice, the development of kidney cysts from triptolide or vehicle-treated animals was analysed.. Although Pkd1 deletion on postnatal Days P10 and P12 resulted in numerous cysts by P35, daily injections with triptolide beginning on Day P16 significantly reduced the total number of cysts per kidney, with a pronounced effect on the number of microcysts and the overall cystic burden. Additionally, renal function as assessed by blood urea nitrogen levels was also improved in triptolide-treated mice at both the P22 and P35 time points. As the Pkd1(flox/flox);Mx1Cre model has not been previously used for drug development studies, the feasibility of a 6-month adult Pkd1 inactivation study was also tested. While kidney cyst formation was minimal and focal in nature, livers of these Pkd1-deficient mice were severely cystic, enlarged and pale.. These results suggest that the Pkd1(flox/flox);Mx1Cre model of ADPKD is amenable to short-term kidney cyst formation drug studies; however, it may be problematic for long-term therapeutic research where widespread liver cysts and fibrosis could compromise drug metabolism.

Topics: Aging; Animals; Antineoplastic Agents, Alkylating; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p21; Disease Models, Animal; Disease Progression; Diterpenes; Epoxy Compounds; GTP-Binding Proteins; Integrases; Kidney; Mice; Mice, Mutant Strains; Mutation; Myxovirus Resistance Proteins; Phenanthrenes; Polycystic Kidney, Autosomal Dominant; Time Factors; TRPP Cation Channels

Spinal cord injury (SCI) is a cause of major neurological disability, and no satisfactory treatment is currently available. Traumatic SCI directly damages the cell bodies and/or processes of neurons and triggers a series of endogenous processes, including neuroinflammatory response and reactive astrogliosis. In this study, we found that triptolide, one of the major active components of the traditional Chinese herb Tripterygium wilfordii Hook F, inhibited astrogliosis and inflammation and promoted spinal cord repair. Triptolide was shown to prevent astrocytes from reactive activation by blocking the JAK2/STAT3 pathway in vitro and in vivo. Furthermore, astrocytic gliosis and glial scar were greatly reduced in injured spinal cord treated with triptolide. Triptolide treatment was also shown to decrease the ED-1 or CD11b-positive inflammatory cells at the lesion site. Using neurofilament staining and anterograde tracing, a significantly greater number of regenerative axons were observed in the triptolide-treated rats. Importantly, behavioral tests revealed that injured rats receiving triptolide had improved functional recovery as assessed by the Basso, Beattie, and Bresnahan open-field scoring, grid-walk, and foot-print analysis. These results suggested that triptolide promoted axon regeneration and locomotor recovery by attenuating glial scaring and inflammation, and shed light on the potential therapeutic benefit for SCI.

Topics: Animals; Animals, Newborn; Astrocytes; Cells, Cultured; Disease Models, Animal; Diterpenes; Epoxy Compounds; Exploratory Behavior; Glial Fibrillary Acidic Protein; Gliosis; Immunosuppressive Agents; Inflammation; Janus Kinase 2; Male; Neurofilament Proteins; Phenanthrenes; Psychomotor Performance; Pyramidal Tracts; Rats; Rats, Sprague-Dawley; Signal Transduction; Spinal Cord Injuries; STAT3 Transcription Factor

Membranous nephropathy is a major cause of nephrotic syndrome in adults where podocyte injuries were found to mediate the development of proteinuria. Triptolide, a major active component of Tripterygium wilfordii Hook F, has potent immunosuppressive, anti-inflammatory and antiproteinuric effects. To study its antiproteinuric properties, we established an experimental rat model of passive Heymann nephritis and a C5b-9 injury model of podocytes in vitro. Treatment or pretreatment with triptolide markedly reduced established proteinuria as well as the titer of circulating rat anti-rabbit IgG antibodies in these nephritic rats, accompanied by a reduction in glomerular C5b-9 deposits. Expression of desmin, a marker of podocyte injury, diminished after triptolide treatment, whereas quantitative analysis of mean foot process width showed that effacement of foot processes was substantially reversed. In in vitro studies we found that triptolide deactivated NADPH oxidase, suppressed reactive oxygen species generation and p38 mitogen-activated protein kinase, and restored RhoA signaling activity. Triptolide did not interfere with the formation of C5b-9 on the membrane of podocytes. Thus, triptolide reduces established heavy proteinuria and podocyte injuries in rats with passive Heymann nephritis, and protects podocytes from C5b-9-mediated injury.

Topics: Administration, Oral; Animals; Cell Line; Complement Membrane Attack Complex; Cytoprotection; Desmin; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Glomerulonephritis, Membranous; Heymann Nephritis Antigenic Complex; Immunoglobulin G; Immunosuppressive Agents; Mice; NADPH Oxidases; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Podocytes; Proteinuria; Rabbits; Rats; Rats, Sprague-Dawley; Rats, Wistar; Reactive Oxygen Species; rho GTP-Binding Proteins; rhoA GTP-Binding Protein; Signal Transduction; Tacrolimus; Time Factors

Triptolide (TPT) is a diterpenoid triepoxide extracted from the Chinese herb Tripterygium wilfordii Hook. F. It exhibits potent immunosuppressive and anti-inflammatory properties. This study was undertaken to investigate its effects on prolongation of islet allograft survival in rodents. Additionally, we investigated whether TPT would be toxic to islet function in vivo.. We transplanted BALB/c islets to either chemically induced diabetic C57BL/6 mice or spontaneously diabetic nonobese diabetic (NOD) mice. TPT was injected within 2 weeks or continuously, until rejection, in the two combinations. Then, we evaluated the toxicity of TPT on islet function by daily injection to naive BALB/c or diabetic BALB/c that was cured by syngeneic islet transplantation under the kidney capsule. Mice injected with cyclosporine A (CsA) or vehicle served as controls. Intraperitoneal glucose tolerance tests (IPGTTs) performed at 4 and 8 weeks in the naive BALB/c group, and at 2, 4, 6, and 8 weeks in the syngeneic transplanted group.. The medium survival time of islets allograft from TPT treated C57BL/6 and NOD recipients were 28.5 days (range 24-30 days, n=10) and 33.0 days (range 15-47 days, n=6), respectively, and they were significantly different from those of the vehicle treated controls, which were 14.0 days (range 13-16 days, n=6) and 5.0 days (range 4-10 days, n=6), respectively (all P<0.0001). The IPGTT demonstrated that there was no difference between the TPT treated and vehicle treated groups, either in the normal or syngeneic transplanted islet BALB/c mice. However, CsA injection impaired islet function in both normal and syngeneic transplanted mice as early as 4 weeks.. TPT prolonged islets allograft survival in a chemically induced diabetic or an autoimmune diabetic murine model without impairment of islet function.

Topics: Animals; Blood Glucose; Cyclosporine; Diabetes Mellitus; Diabetes Mellitus, Experimental; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Glucose Tolerance Test; Graft Rejection; Graft Survival; Immunosuppressive Agents; Islets of Langerhans; Islets of Langerhans Transplantation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred NOD; Phenanthrenes; Time Factors; Transplantation, Homologous; Weight Gain

Several lines of research suggest that estrogens (and estrogenic compounds) are neuroprotective following experimental traumatic brain injury. However, therapeutic use of estrogens in this and other regards remains controversial. Therefore, analysis of estrogen-like compounds without potential problems similar to estrogens seems warranted. (±) Z-Bisdehydrodoisynolic acid (Z-BDDA) is a seco-steroid that has potent estrogenic as well as antioxidant activities in vitro and in vivo. We evaluated the therapeutic potential of Z-BDDA (300μg/0.1cc/100g body weight, sc) to promote the recovery of behavioral function following lateral fluid percussion injury (FPI) to the brain in male rats. Two hours subsequent to FPI, treatment with Z-BDDA began with a bolus subcutaneous (sc) injection followed by booster treatments given 24 and 48h later. Behavioral testing was initiated on the second day after FPI and results of Z-BDDA treatments were compared to treatment with vehicle only and to sham FPI surgery. Z-BDDA effectively enhanced recovery of coordinated limb movement assessed by locomotor placing performance across the duration of the study. Z-BDDA treated animals also performed better on a spatial memory task in the Morris water maze, showing improved learning curves across days of testing. Vestibulomotor function, measured by beam walk performance, appeared to improve in Z-BDDA treated animals, however these results did not reach statistical significance (p>0.05). Following cessation of the behavioral testing, all animals underwent assessments of gross neuroanatomical pathology. Cortical lesion size and cell death analysis with Fluoro-jade B failed to reveal Z-BDDA enhanced neuroprotection. These findings support our hypothesis that Z-BDDA can facilitate behavioral recovery following FPI in adult male rats although the mechanism(s) of these effects remain to be determined.

Topics: Animals; Brain Injuries; Disease Models, Animal; Estrogens; Male; Phenanthrenes; Rats; Rats, Long-Evans; Recovery of Function; Secosteroids

To investigate the underlying neuroprotective mechanisms of Tanshinone II A (TSA) on rat cerebral ischemia in vivo.. Study of TSA on rat cerebral ischemia in vivo: Male SD rats were divided into four groups (sham-operated, ischemic and treated group (lower dose and higher dose). Chronic cerebral ischemmia after permanent bilateral carotid artery ligation was introduced as an in vivo ischemic model. After ischemia impairment, TSA (2, 4 mg x kg(-1) x d(-1)) was administrated by ip for 30 days in treated group. We used Morris water maze to investigate the learning and memory. Levels of malondialdehyde (MDA), activity of superoxide dismetase (SOD) and glutathione peroxidase (GPX) in brain tissue were detected by spectrophotometer. High-performance liquid chromatography (HPLC) with fluorescence detection was applied to measure the contents of glutamate and gamma-aminobutyric acid (GABA) in cortex and hippocampus.. TSA can improve learning and memory deficits in vascular dementia. An elevation of SOD and GPX activity and decrease of MDA level were shown in TSA treated group after brain ischemia. Decreased glutamate and gamma-aminobutyric acid induced by chronic brain ischemia were markedly inhibited by TSA pretreatment.. The neuroprotective effect of TSA are partly due to its functions as follow: anti-free radical injury; regulating the content of glutamate and gamma-aminobutyric acid.

Topics: Abietanes; Animals; Dementia, Vascular; Disease Models, Animal; Drugs, Chinese Herbal; Humans; Male; Malondialdehyde; Maze Learning; Memory; Neuroprotective Agents; Phenanthrenes; Random Allocation; Rats; Rats, Sprague-Dawley

Chronic kidney disease (CKD) is a common cause of end-stage renal disease. Antihypertensive agents are used clinically to inhibit the progression of CKD, but cannot prevent eventual renal failure. This study investigated the effect of Tanshinone IIA, an active component of Salvia miltiorrhiza, in rats suffering from CKD induced by 5/6 nephrectomy. After development of renal insufficiency, the rats were treated with Tanshinone IIA (10 mg/kg) for 8 weeks. Serum creatinine, angiotensin II (Ang II), transforming growth factor β1 (TGF-β1) and collagen IV levels were significantly reduced in Tanshinone IIA treated rats compared with a control group. In addition, Tanshinone IIA suppressed increases in urinary protein excretion in CKD rats. These findings suggest that chronic oral administration of Tanshinone IIA can improve renal dysfunction associated with CKD.

Topics: Abietanes; Administration, Oral; Angiotensin II; Animals; Collagen Type IV; Creatinine; Disease Models, Animal; Drugs, Chinese Herbal; Kidney; Male; Phenanthrenes; Proteinuria; Rats; Rats, Sprague-Dawley; Renal Insufficiency, Chronic; Salvia miltiorrhiza; Transforming Growth Factor beta1

To observe the prevention and therapeutic effects of tanshitone (TAN) on retinoic acid induced osteoporosis in mice.. The mice osteoporosis was induced by given retinoic acid intragastrically for two weeks. The histomorphological features of bone were observed and biochemical indexes in serum (Ca, P, ALP, TRAP, E2, BGP) were determined after the mice were given TAN at the dose of 40, 80, 160 mg x kg(-1) respectly.. Tanshinone can induce high conversion of osteoporosis. The levels of P, ALP, TRAP and BGP in the TAN groups were lower than the model group, while the E2 level was higher than the model group.. Tanshitone can prevent the loss bone in the experimental mice. The mechanism may be that it improves the level of estrogenic hormone and inhibits the high bone turnover.

Topics: Abietanes; Alkaline Phosphatase; Animals; Bone Density; Disease Models, Animal; Drugs, Chinese Herbal; Female; Humans; Male; Mice; Osteoporosis; Phenanthrenes; Tretinoin

Tripterygium wilfordii Hook. f. (Celastraceae) has been traditionally used as folk medicine for centuries in China for the treatment of immune-inflammatory diseases.. This study aimed to assess the antiangiogenic activities which support the therapeutic use of Tripterygium wilfordii and its terpenoids for angiogenesis disease such as cancer.. The ethanol extract of Tripterygium wilfordii and subsequent fractions were evaluated on an in vivo antiangiogenic zebrafish embryo model.. Three antiangiogenic terpenoids were isolated by bioassay-guided purification, namely, celastrol (4), cangoronine (5) and triptolide (7). Among them, triptolide manifested the most potent antiangiogenic activity against vessel formation by nearly 50% at 1.2 microM. Semi-quantitative RT-PCR analysis revealed that triptolide dose- and time-dependently reduced the mRNA expression of angiopoietin (angpt)2 and tie2 in zebrafish, indicating the involvement of angpt2/tie2 signaling pathway in the antiangiogenic action of triptolide.. The discovery of an alternative pathway further confirms the value of ethnopharmacological investigations into traditional botanicals for leads for potential drug development.

Topics: Angiogenesis Inhibitors; Animals; Disease Models, Animal; Diterpenes; Drugs, Chinese Herbal; Embryo, Nonmammalian; Epoxy Compounds; Neovascularization, Physiologic; Pentacyclic Triterpenes; Phenanthrenes; Plant Roots; Reverse Transcriptase Polymerase Chain Reaction; Terpenes; Tripterygium; Triterpenes; Zebrafish

The amyloid precursor protein (APP) is cleaved enzymatically by non-amyloidogenic and amyloidogenic pathways. alpha-Secretase cleaves APP within beta-amyloid protein (Abeta) sequence, resulting in the release of a secreted fragment of APP (sAPPalpha) and precluding Abeta generation. Cryptotanshinone (CTS), an active component of the medicinal herb Salvia miltiorrhiza, has been shown to improve learning and memory in several pharmacological models of Alzheimer's disease (AD). However, the effects of CTS on the Abeta plaque pathology and the APP processing in AD are unclear. Here we reported that CTS strongly attenuated amyloid plaque deposition in the brain of APP/PS1 transgenic mice. In addition, CTS significantly improved spatial learning and memory in APP/PS1 mice assessed by the Morris water maze testing. To define the exact molecular mechanisms involved in the beneficial effects of CTS, we investigated the effects of the CTS on APP processing in rat cortical neuronal cells overexpressing Swedish mutant human APP695. CTS was found to decrease Abeta generation in concentration-dependent (0-10muM) manner. Interestingly, the N-terminal APP cleavage product, sAPPalpha was markedly increased by CTS. Further study showed that alpha-secretase activity was increased by CTS. Taken together, our results suggested CTS improved the cognitive ability in AD transgenic mice and promoted APP metabolism toward the non-amyloidogenic products pathway in rat cortical neuronal cells. CTS shows a promising novel way for the therapy of AD.

Topics: Alzheimer Disease; Amyloid beta-Peptides; Amyloid beta-Protein Precursor; Amyloid Precursor Protein Secretases; Animals; Brain; Camphanes; Cerebral Cortex; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Memory Disorders; Mice; Mice, Inbred C57BL; Mice, Transgenic; Panax notoginseng; Phenanthrenes; Plaque, Amyloid; Presenilin-1; Rats; Salvia miltiorrhiza; Up-Regulation

Triptolide (TPL), a diterpenoid triepoxide purified from the Chinese herb Tripterygium wilfordii Hook F, has been reported to potentiate the anti-tumor effect in various cancer cells. However, the effect of TPL on oral cancers is not yet evaluated. Herein we first demonstrate that TPL induces prominent growth inhibition and apoptosis in two oral cancer cell lines, SCC25 and OEC-M1 and in KB cells. Our results indicate that TPL induces a dose-dependent apoptosis of these cells at nanomolar concentration. Apoptosis signalings are both activated through time upon TPL treatment detected by elevated caspase-3, 8, 9 activities. In xenograft tumor mouse model, TPL injection successfully inhibits the tumor growth via apoptosis induction which was demonstrated by TUNEL assay. These results demonstrate that TPL exerts anti-tumor effect on oral cancer and KB cells and suggest further the potential of TPL combining with other chemotherapeutic agents or radiotherapy for advanced oral cancer.

Topics: Animals; Antineoplastic Agents; Apoptosis; Carcinoma, Squamous Cell; Disease Models, Animal; Diterpenes; Drug Screening Assays, Antitumor; Epoxy Compounds; Humans; KB Cells; Mice; Mice, Inbred NOD; Mice, SCID; Mouth Neoplasms; Phenanthrenes

Phenanthrene imidazoles 26 and 44 have been identified as novel potent, selective and orally active mPGES-1 inhibitors. These inhibitors are significantly more potent than the previously reported chlorophenanthrene imidazole 1 (MF63) with a human whole blood IC50 of 0.20 and 0.14 microM, respectively. It exhibited a significant analgesic effect in a guinea pig hyperalgesia model at oral doses as low as 14 mg/kg. Both active and selective mPGES-1 inhibitors (26 and 44) have a relatively distinct pharmacokinetic profile and are suitable for clinical development.

Topics: Administration, Oral; Animals; Benzimidazoles; Disease Models, Animal; Drug Design; Enzyme Inhibitors; Guinea Pigs; Humans; Hyperalgesia; Intramolecular Oxidoreductases; Nitriles; Phenanthrenes; Prostaglandin-E Synthases; Recombinant Proteins; Structure-Activity Relationship

Available treatments for multiple sclerosis (MS) require frequent injections and have significant side effects. In this study, we examined the immunomodulatory properties of orally administered triptolide, a major diterpenoid triepoxide isolated from a twining vine Tripterygium wilfordii. SJL/J mice were primed with PLP(139-151) peptide and orally treated with triptolide (100mug/kg per day) from the day of EAE induction (preventive regime) and after the onset of clinical signs (therapeutic regime). Triptolide delayed disease onset, reduced clinical symptoms, decreased the relapse rate, and suppressed inflammation and demyelination in CNS tissue of EAE mice when compared to vehicle-treated animals. Molecular analysis revealed a marked increase of heat shock protein 70 (Hsp70) mRNA and protein in the CNS tissue of triptolide-treated animals. Cytokine and chemokine expression analysis from EAE tissues and in vitro macrophages detected a decrease of key pro-inflammatory mRNAs. Triptolide inhibited IkappaBalpha phosphorylation and NF-kappaB nuclear translocation by stabilization of NF-kappaB/IkappaBalpha complex, possibly due to a direct physical interaction between NF-kappaB and Hsp70 proteins. Lymph node cell proliferation assay in EAE confirmed the immunosuppressive efficacy of triptolide. Our data indicate that daily oral administration of triptolide exhibits not only a preventive but also a therapeutic effect on EAE. These effects might be explained by the increase in Hsp70 levels driven by triptolide and stabilization of the NF-kappaB/IkappaBalpha complex leading to an attenuated inflammatory response.

Topics: Administration, Oral; Analysis of Variance; Animals; Cell Line, Transformed; Cytokines; Cytoplasm; Disease Models, Animal; Diterpenes; Encephalomyelitis, Autoimmune, Experimental; Epoxy Compounds; HSP70 Heat-Shock Proteins; I-kappa B Proteins; Immunoprecipitation; Immunosuppressive Agents; Lymph Nodes; Macrophages; Mice; Myelin Proteolipid Protein; NF-kappa B; NF-KappaB Inhibitor alpha; Peptide Fragments; Phenanthrenes; Phosphorylation; Phytotherapy; RNA, Messenger; Signal Transduction; Time Factors; Tripterygium; Up-Regulation

Triptolide and tripdiolide are thought to be active components of the Chinese antirheumatic herbal remedy Tripterygium wilfordii Hook F, which has been shown to be effective in treating murine lupus nephritis. This study was undertaken to examine the therapeutic effect of triptolide and tripdiolide on established lupus nephritis in (NZB x NZW)F1 mice.. (NZB x NZW)F1 mice were treated with vehicle, triptolide, or tripdiolide for 15 weeks beginning at the age of 29 weeks (after the development of lupus nephritis). Body weight, proteinuria, and anti-double-stranded DNA (anti-dsDNA) antibodies were monitored, and the kidney and spleen were assessed histologically. Culture supernatants of spleen mononuclear cells were assayed for cytokines.. By 28 weeks, most (NZB x NZW)F1 mice had developed lupus nephritis. Vehicle-treated mice exhibited progressive proteinuria, hypoalbuminemia, elevated blood urea nitrogen (BUN) levels, and evidence of severe nephritis. In contrast, proteinuria and BUN levels were significantly reduced in mice treated with either triptolide or tripdiolide as compared with those treated with vehicle. There was no hypoalbuminemia or apparent evidence of lupus nephritis in mice treated with either of the 2 diterpenoids. At 44 weeks of age, the survival rate in mice treated with vehicle (35.7%) was markedly lower than that in mice treated with either triptolide (87.5%) or tripdiolide (88.2%). The mean level of anti-dsDNA antibody in mice treated with tripdiolide was lower than that in the vehicle-treated mice upon completion of the treatment course. Production of tumor necrosis factor, interleukin-6, and monocyte chemoattractant protein 1 by spleen cells was also decreased after diterpenoid therapy.. Therapy with triptolide or tripdiolide significantly ameliorated lupus nephritis in (NZB x NZW)F1 mice, reduced cytokine and chemokine production, and prolonged survival.

Topics: Animals; Disease Models, Animal; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Female; Immunosuppressive Agents; Lupus Nephritis; Mice; Phenanthrenes; Spleen; Tripterygium

This study examined the effects of tanshinone derivatives (tanshinone I, cryptotanshinone, 15,16-dihydrotanshinone I) on prostaglandin (PG) and nitric oxide (NO) metabolism in an attempt to establish their anti-inflammatory mechanisms and to present a scientific rationale for the use of Salvia miltiorrhiza (danshen) in inflammatory conditions. From lipopolysaccharide-treated RAW 264.7 cells, cyclooxygenase-2 (COX-2)-mediated PGE2 production was inhibited by tanshinone I, cryptotanshinone and 15,16-dihydrotanshinone I, while only cryptotanshinone and 15,16-dihydrotanshinone I inhibited inducible NO synthase (iNOS)-mediated NO synthesis at 1-50 microM. Particularly, cryptotanshinone was found to be a down-regulator of proinflammatory molecule expression, including COX-2 and iNOS. The electrophoretic mobility shift assay showed that cryptotanshinone and 15,16-dihydrotanshinone I also inhibited the activation of the transcription factors, such as nuclear transcription factor-kappaB and activator protein-1. Moreover, cryptotanshinone exhibited in vivo anti-inflammatory activity against carrageenan-induced paw edema in rats. Overall, these results provide additional scientific rationale for the anti-inflammatory use of danshen in Chinese medicine. Especially, cryptotanshinone and 15,16-dihydrotanshinone I are important constituents.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Carrageenan; Cell Line; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Down-Regulation; Edema; Electrophoretic Mobility Shift Assay; Furans; Lipopolysaccharides; Macrophages; Male; Mice; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Phenanthrenes; Plant Roots; Quinones; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza

Triptolide, the principal active ingredient in the extract of Chinese herb Tripterygium wilfordii Hook , has both anti-inflammatory and immunomodulatory activities. However, the potential therapeutic role of triptolide in IBD was still unknown. Interleukin-10 deficient mice, a well characterized experimental model of inflammatory bowel disease, spontaneously developed a Th1 T cell-mediated colitis with many similarities to Crohn's disease. This study was designed to investigate the therapeutic effect of triptolide on the chronic colitis in IL-10-/- mice.. Triptolide was intraperitoneally administrated every another day for 8 weeks to IL-10-/- mice. The gross and histological appearances of the colon, the level of inflammatory mediators and transcription factor activation in the colon were evaluated and compared with the control group.. The 8-week administration of triptolide resulted in a significant decrease in the severity of colitis, together with lower production of TNF-alpha ,IFN-gamma and IL-4 in colon. The level of serum amyloid A was decreased in triptolide-treated mice. Gene expressions of IL-12 and IL-23 in colon were also downregulated after treatment. Furthermore, administration of triptolide markedly reduced NF-small ka, CyrillicB activation in colon mucosa of IL-10-/- mice.. The efficacy of tritpolide treatment for the reduction of intestinal inflammation in IL-10-/- mice is a result of both anti-inflammatory and immunosuppressive activity. Triptolide holds significant potential for clinical applications for CD treatment.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Chronic Disease; Colitis; Colon; Disease Models, Animal; Diterpenes; Epoxy Compounds; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-23; Interleukin-4; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; NF-kappaB-Inducing Kinase; Phenanthrenes; Protein Serine-Threonine Kinases; Serum Amyloid A Protein; Tumor Necrosis Factor-alpha

To explore the molecular biological mechanism of tanshinone II A (TSN) in preventing hypertensive left ventricular hypertrophy (HLVH) through studying the effects of TSN on angiotensin receptor (ATR) expression and free calcium ion ([Ca2+]i) in rats with hypertrophic myocardium caused by abdominal aorta constriction.. SD rats were established into HLVH model by abdominal aorta constriction operation, they were randomly divided into the model group, the three treated groups treated respectively with intra peritoneal injection of low dose TSN (10 mg/kg) and high dose TSN (20 mg/kg) and gastrogavage of Valsartan (10 mg/kg) once a day 4 weeks after modeling. Besides, 8 sham-operated SD rats were set up as the control group. Eight weeks later, rats' caudal arterial pressure was measured, and their hearts were taken for measuring the left ventricular mass index (LVMI) and myocardial fiber diameter (MFD) by HE stain of the pathological section. Moreover, the mRNA and protein expressions of AT1 and AT2 receptors in the left ventricular tissue were detected by RT-PCR and Western blot, and [Ca2+]i concentration was determined with laser-scanning confocal microscope.. (1) The elevated blood pressure in the TSN treated groups, either high or low dose, remained unchanged, significantly higher than that in the control group and the Valsartan treated group (P < 0.01, P < 0.05). (2) LVMI and MFD in the three treated groups were significantly lower than those in the model group (P <0.01), respectively, although they were higher than those in the control group (P <0.05). (3) The mRNA and protein expressions of AT1 receptor were obviously lower in the three treated groups than those in the model groups (P < 0.05); but the lowering was more significant in the valsartan treated group (P < 0.05). (4) The mRNA and protein expressions of AT2 receptor were significantly higher in the Valsartan treated group as compared with other groups (P < 0.05), while the difference among the other groups showed no statistical significance (P > 0.05). (5) The elevated (Ca2+]i concentration in hypertrophic myocardium after modeling was significantly lowered after treatment in the three treated groups (P < 0.05), but the lowering in the high TSN treated group was more significant than that in the Valsartan treated group (P <0.05).. The inhibition of TSN on myocardial hypertrophy is blood pressure independent, its mechanism is possibly related with the inhibition on AT1R gene expression and the blocking of free calcium ion influx in cardiac muscle cells. AT2 receptor may participate the effect of Valsartan in lowering blood pressure and reversing myocardial hypertrophy.

Topics: Abietanes; Animals; Blood Pressure; Calcium; Cardiomyopathy, Hypertrophic; Disease Models, Animal; Female; Humans; Male; Myocardium; Phenanthrenes; Random Allocation; Rats; Rats, Sprague-Dawley; Receptors, Angiotensin

Tanshinone IIA isolated from Danshen is widely used in Oriental medicine. However, the action of tanshinone IIA in inflammatory bone-resorptive diseases remains unknown. Here we examined the effect of tanshinone IIA in inflammation-mediated osteoclastic bone resorption. Tanshinone IIA inhibited osteoclast differentiation in cocultures of bone marrow cells and calvarial osteoblasts. Tanshinone IIA regulated the expression of receptor activator of NF-kappaB ligand and osteoprotegerin in osteoblasts treated with lipopolysaccharide (LPS). Also, tanshinone IIA inhibited prostaglandin E(2) (PGE(2)) synthesis by inhibiting Cyclooxygenase-2 (COX-2) expression induced by LPS. Furthermore, tanshinone IIA greatly suppressed bone loss in the mouse models of bone loss. Our findings suggest that tanshinone IIA inhibits osteoclast formation by inhibiting COX-2/PGE(2) signaling and by suppressing bone erosion in vivo. These results suggest that tanshinone IIA may be of therapeutic value as an anti-bone-resorptive drug in the treatment of bone-related disease.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Bone Diseases; Bone Marrow Cells; Bone Resorption; Cell Differentiation; Cyclooxygenase 2; Dinoprostone; Disease Models, Animal; Drugs, Chinese Herbal; Gene Expression Regulation; Inflammation; Male; Mice; Mice, Inbred ICR; Osteoblasts; Osteoclasts; Phenanthrenes

To determine the influence of tanshinone on the levels of nitric oxide synthase (NOS) and acetylcholinesterase (AChE) in the brain of an Alzheimer's Disease (AD) rat model and on its potential therapeutic mechanism.. 100 Male Sprague Dawley rats were divided into three groups: control group, model group and tanshinone treatment group. 10 microg A beta 1-42 was injected bilaterally into the dorsal lateral region of the dentate gyrus in the hippocampus of rats in the model and tanshinone treatment groups to prepare the AD models. 24h after modeling, tanshinone, 50mg/kg, was administered by gastric perfusion to rats in the tanshinone treatment group. Later, immunohistochemical assay and Western blot analysis were used to detect expression of neuronal NOS (nNOS) and inducible NOS (iNOS) in the rat hippocampus. Activity of AChE in each subregion (CA1 approximately CA4) of rats' hippocampus was determined by a histochemical technique.. Expression of nNOS in the model group was down-regulated whereas iNOS was up-regulated. After A beta 1-42 injection, the number of AChE positive fibers in each subregion (CA1 approximately CA4) of the hippocampus was decreased compared with controls. With tanshinone administration, the changes were improved to varying degrees.. Tanshinone modulates AChE and NOS proteins concentrations in the hippocampus of AD rats. This may have therapeutic potential in AD rats.

Topics: Abietanes; Acetylcholinesterase; Alzheimer Disease; Animals; Disease Models, Animal; Gene Expression Regulation, Enzymologic; Hippocampus; Immunosuppressive Agents; Male; Nitric Oxide Synthase Type I; Nitric Oxide Synthase Type II; Phenanthrenes; Rats; Rats, Sprague-Dawley

Tanshinone IIA (Tan-IIA) was isolated from Salviae Miltiorrhizae Radix. Our previous studies showed that Tan-IIA induced apoptosis in human colon cancer colo 205 cells, but the molecular mechanisms of the effect of Tan-IIA on human colon cancer were not clearly elucidated. The protein expression of ErbB-2 was up-regulated and activated in human and experimental colon cancers. In the present study, the effects of Tan-IIA on the protein expression of ErbB-2 in colo 205 cells were investigated. In vitro, colo 205 cells were treated with various concentrations of Tan-IIA (1, 2 and 5 mug/ ml) for 24 h, and the protein expression of TNF-alpha, ErbB-2 and caspase-3 was assayed by Western blotting. For the in vivo studies, male SCID mice were xenografted with colo 205 cells, and from day 10, Tan IIA (20 mg/kg/day, dissolved in corn oil) was administered by oral feeding for 30 days. As a control, mice with xenografted tumors were separately treated with corn oil (0.1 ml/10 g body weight). Expression of TNF-alpha, ErbB-2 and caspase-3 proteins was measured by Western blot analysis. Our results showed that Tan-IIA down-regulated the protein expression of ErbB-2 and up-regulated TNF-alpha and caspase-3 in colo 205 cells in vitro. In a colo 205 xenograft model, treatment with Tan-IIA caused up-regulation of TNF-alpha, caspase-3 and down-regulation of ErbB-2 protein expression as compared to the controls. Based on these observations, one possible molecular mechanisms by which Tan-IIA inhibits the proliferation of colo 205 cells is through the down-regulation of ErbB-2 protein expression and the up-regulation of the protein expression of TNF-alpha and caspase-3.

Topics: Abietanes; Actins; Animals; Antineoplastic Agents, Phytogenic; Caspase 3; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; Disease Models, Animal; Down-Regulation; Humans; Male; Mice; Mice, SCID; Neoplasm Transplantation; Phenanthrenes; Receptor, ErbB-2; Tumor Necrosis Factor-alpha; Up-Regulation

To study the effects of cryptotanshinone on androgen synthesis for the prenatally androgenized male rats.. On days 16-18 of pregnancy, rats were injected s. c. with testosterone propionas continuously for 3 days; male offspring were studied as subject. Serum concentrations of testosterone (T), 17a-hydroxy progesterone (17-OHP), blood glucose, and insulin were measured by radioimmunoassay. Then, the rats were treated with cryptotanshinone by gavage for 14 days, and the levels of serum T, 17-OHP and insulin were detected and the 17a-hydroxylase protein expression in interstitial cell was measured using the method of immunohistochemistry.. There was no difference between the male groups who were prenatally androgenized in serum levels of T, but the 17-OHP, fasting insulin levels and homeostatic model assessment for insulin resistance (HOMA-IR) elevated significantly (P < 0.05). Cryptotanshinone could lower the levels of 17-OHP (P < 0.05) but had no effect on 17a-hydroxylase.. Prenatally androgenized male rats exhibit elevated 17-OHP and diminished insulin sensitivity. Cryptotanshinone could decrease 17-OHP, but has no effect on insulin, indicating it may reduce androgen synthesis.

Topics: Androgens; Animals; Disease Models, Animal; Female; Humans; Male; Maternal Exposure; Phenanthrenes; Polycystic Ovary Syndrome; Pregnancy; Prenatal Exposure Delayed Effects; Random Allocation; Rats; Rats, Wistar

Tanshinone IIA (Tan IIA) is a member of the major lipophilic components abstracted from the root of Salvia miltiorrhiza Bunge and has the capacity of anti-atherosclerosis. To investigate the potential mechanism, we established an animal model by giving high fatty diet to rabbits and Tan IIA was given in different dose. Then, superoxide dismutase (SOD) activity and the malondialdehyde (MDA) level in serum were detected using spectrophotometry; cluster of differentiation 40 (CD40) expression of cellular membrane fraction of aortas and matrix metalloproteinase-2 (MMP-2) activity of total protein extract of aortas were detected by Western Blotting and Zymography, respectively. Compared with the control group, the level of MDA, the expression of CD40 and the MMP-2 activity were increased while the SOD activity was decreased significantly in model group. After Tan IIA administration, the SOD activity was significantly increased while the level of MDA was decreased; both the expression of CD40 and the activity of MMP-2 were decreased. It is suggested that Tan IIA not only inhibits the oxidation but also suppresses the inflammation in atherosclerotic lesion. Our data suggest that not only anti-oxidation but also anti-inflammation by decrease the expression of CD40 and MMP-2 activity maybe the potential mechanisms by which Tan IIA anti-atherosclerosis.

Topics: Abietanes; Animals; Anti-Inflammatory Agents; Antioxidants; Atherosclerosis; CD40 Antigens; Dietary Fats; Disease Models, Animal; Dose-Response Relationship, Drug; Down-Regulation; Female; Male; Malondialdehyde; Matrix Metalloproteinase 2; Phenanthrenes; Plant Roots; Rabbits; Salvia miltiorrhiza; Superoxide Dismutase

Atherosclerosis is the leading cause of death in Western societies and a chronic inflammatory disease. However, the key mediators linking recruitment of inflammatory cells to atherogenesis remain poorly defined. Poly(ADP-ribose) polymerase 1 (PARP1) is a nuclear enzyme, which plays a role in acute inflammatory diseases.. In order to test the role of PARP in atherogenesis, we applied chronic pharmacological PARP inhibition or genetic PARP1 deletion in atherosclerosis-prone apolipoprotein E-deficient mice and measured plaque formation, adhesion molecules, and features of plaque vulnerability. After 12 weeks of high-cholesterol diet, plaque formation in male apolipoprotein E-deficient mice was decreased by chronic inhibition of enzymatic PARP activity or genetic deletion of PARP1 by 46 or 51%, respectively (P < 0.05, n >or= 9). PARP inhibition or PARP1 deletion reduced PARP activity and diminished expression of inducible nitric oxide synthase, vascular cell adhesion molecule-1, and P- and E-selectin. Furthermore, chronic PARP inhibition reduced plaque macrophage (CD68) and T-cell infiltration (CD3), increased fibrous cap thickness, and decreased necrotic core size and cell death (P < 0.05, n >or= 6).. Our data provide pharmacological and genetic evidence that endogenous PARP1 is required for atherogenesis in vivo by increasing adhesion molecules with endothelial activation, enhancing inflammation, and inducing features of plaque vulnerability. Thus, inhibition of PARP1 may represent a promising therapeutic target in atherosclerosis.

Topics: Animals; Apolipoproteins E; Atherosclerosis; Cell Adhesion Molecules; Cholesterol; Disease Models, Animal; E-Selectin; Enzyme Inhibitors; Inflammation; Inflammation Mediators; Macrophages; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Necrosis; Nitric Oxide Synthase Type II; P-Selectin; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; T-Lymphocytes; Vascular Cell Adhesion Molecule-1

Inhaled airborne pollutants such as particulate matter increase the susceptibility to adverse health consequences and cardiovascular events. Diesel exhaust contributes significantly to the ambient particle pollution burden. The purpose of this investigation was to determine if exposure to a common component of diesel exhaust, phenanthraquinone (PQ), impairs endothelium-dependent vasodilation of the femoral principal nutrient artery and to determine whether age, gender, and/or hormonal status alter the putative effects of PQ on vasodilation. Vasodilation to acetylcholine (ACh) was assessed in vitro in intact control (age 6, 14, and 24 mo) and ovariectomized (age 6, 14, and 24 mo) female rats and intact (age 6 and 24 mo) male rats. Gender did not influence vasodilator capacity of the femoral principal nutrient artery, and there was an age-related decline in endothelium-dependent vasodilation in both female and male 24-mo-old rats. Exposure to PQ elicited a gender-specific affect in 6-mo-old rats; i.e., vasodilation was impaired 63% in male rats but had no effect in female rats. Exposure to PQ abolished vasodilation in 14- and 24-mo-old rats of both genders, and ovariectomy compromised vasodilator responsiveness to ACh in all age groups. The data demonstrate a vasoprotective mechanism in young female rats that may be related to endogenous ovarian hormones and provides evidence that suggests certain subsets of the population (e.g., elderly, males, and postmenopausal women) may be more susceptible to the adverse consequences of airborne pollutants.

Topics: Age Factors; Air Pollutants; Animals; Disease Models, Animal; Endothelium, Vascular; Female; Femoral Artery; Male; Phenanthrenes; Rats; Rats, Inbred F344; Sex Factors; Vasodilation; Vehicle Emissions

The main objective of the present study was to evaluate the reduction in halofantrine (Hf) toxicity, an antimalarial drug frequently associated with QT interval prolongation in electrocardiogram, by its entrapment in poly-epsilon-caprolactone nanocapsules (NC). The acute lethal dose (LD(100)) of Hf.HCl experimentally observed was 200 mg/kg whereas the calculated LD(50) was 154 mg/kg. In contrast, the LD(100) for Hf-NC was 300 mg/kg with a longer mean time to death than Hf.HCl. The calculated LD(50) was 249 mg/kg for Hf-NC. The Hf entrapped in PCL NC presented a greater efficacy than PLA-PEG NC and than Hf solution in P. berghei-infected mice at 1 mg/kg. The cardiovascular parameters, ECG and arterial blood pressure, were evaluated in anaesthetized Wistar rats after the IV administration of a single, especially high dose (100 and 150 mg/kg) of halofantrine base loaded-nanocapsules (Hf-NC) or halofantrine chlorhydrate (Hf.HCl) solution. It was observed that Hf solution caused prolongation of the QT and PR intervals of the ECG; however, this effect was significantly (P<0.001) reduced when Hf was administered entrapped in nanocapsules. The treatment with Hf.HCl induced a pronounced bradycardia and severe hypotension leading to death. The effect of Hf-NC upon heart rate was reduced from 58 to 75% for 100 and 150 mg/kg, respectively, when compared with Hf.HCl solution. These findings show that the encapsulation of halofantrine reduces the QT interval prolongation of ECG in rats and suggest that a modification of drug distribution was possible by using nanocapsules. Hf encapsulation was the main factor responsible for the significant reduction in cardiac toxicity observed.

Topics: Animals; Antimalarials; Bradycardia; Disease Models, Animal; Dose-Response Relationship, Drug; Electrocardiography; Heart; Hypotension; Lethal Dose 50; Long QT Syndrome; Malaria; Male; Mice; Nanocapsules; Phenanthrenes; Plasmodium berghei; Rats; Rats, Wistar

We have previously proved that oxidized low-density lipoprotein (oxLDL), a proatherogenic lipoprotein, plays a pivotal role in the development of atherosclerotic calcification (AC). The present study was performed to investigate whether tanshinone II A (TS II A), an anti-oxidant which has been shown to inhibit in vitro oxidation of LDL, has the effects to inhibit AC in rat model and by which, if any, mechanisms.. Rat AC model was induced by excessive vitamin D(2) (VD) and high cholesterol diet (HCD), which was proven to be successful histopathologically and biochemically.. Administration of AC rats with TS II A (35, 70 mg/kg) dose-dependently attenuated the AC pathological changes, meanwhile reduced the vessel contents of lipid and calcium. However, TS II A had no effects on serum levels of lipids, calcium and 25-OH VD. Further studies revealed that TS II A decreased serum concentration of oxLDL, reduced the superoxide anion production and malondialdehyde (MDA) in vessel. In addition, TS II A increased vessel Cu/Zn SOD activity, upregulated vessel mRNA and protein expression of Cu/Zn SOD.. The results suggested that TS II A significantly attenuated the AC in rat model, which might be attributed to its inhibition of oxLDL production independent of the serum levels of lipids, calcium and 25-OH VD, and that increasing of Cu/Zn SOD activity as well as mRNA and protein expression by TS II A might protect LDL against oxidation induced by superoxide anion in vessel.

Topics: Abietanes; Animals; Antioxidants; Aorta, Thoracic; Atherosclerosis; Calcinosis; Calcium; Cholesterol; Cholesterol, Dietary; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme Induction; Ergocalciferols; Lipoproteins, LDL; Male; Malondialdehyde; Oxidative Stress; Phenanthrenes; Rats; Rats, Sprague-Dawley; RNA, Messenger; Superoxide Dismutase; Superoxides; Time Factors

Tanshinone IIB (TSB) is a major active constituent of the root of Salvia miltiorrhiza (Danshen) used in the treatment of acute stroke. Danshen extracts and TSB have shown marked neuron-protective effects in mouse studies but there is a lack of clinical evidence for the neuron-protective effects of Danshen and its active ingredients. This study investigated the neuron-protective effects of TSB in experimentally stroked rats. TSB at 5 and 25 mg/kg by intraperitoneal injection significantly reduced the focal infarct volume, cerebral histological damage and apoptosis in rats subjected to middle cerebral artery occlusion (MCAO) compared to MCAO rats receiving vehicle. This study demonstrated that TSB was effective in reducing stroke-induced brain damage and may represent a novel drug candidate for further development. Further mechanistic studies are needed for the neuron-protective activity of TSB.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Brain; Brain Infarction; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Infarction, Middle Cerebral Artery; Injections, Intraperitoneal; Male; Nerve Degeneration; Neurons; Neuroprotective Agents; Phenanthrenes; Plant Extracts; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza; Stroke; Treatment Outcome

Cryptotanshinone (CTS), a major constituent from the roots of Salvia miltiorrhiza (Danshen), is widely used in the treatment of coronary heart disease, stroke and less commonly Alzheimer's disease. Our recent study indicates that CTS is a substrate for P-glycoprotein (PgP/MDR1/ABCB1). This study has investigated the nature of the brain distribution of CTS across the brain-blood barrier (BBB) using several in vitro and in vivo rodent models. A polarized transport of CTS was found in rat primary microvascular endothelial cell (RBMVEC) monolayers, with facilitated efflux from the abluminal side to luminal side. Addition of a PgP (e.g. verapamil and quinidine) or multi-drug resistance protein 1/2 (MRP1/2) inhibitor (e.g. probenecid and MK-571) in both luminal and abluminal sides attenuated the polarized transport. In a bilateral in situ brain perfusion model, the uptake of CTS into the cerebrum increased from 0.52 +/- 0.1% at 1 min to 11.13 +/- 2.36 ml/100 g tissue at 30 min and was significantly greater than that of sucrose. Co-perfusion of a PgP/MDR1 (e.g. verapamil) or MRP1/2 inhibitor (e.g. probenecid) significantly increased the brain distribution of CTS by 35.1-163.6%. The brain levels of CTS were only about 21% of those in plasma, and were significantly increased when coadministered with verapamil or probenecid in rats. The brain levels of CTS in rats subjected to middle cerebral artery occlusion and rats treated with quinolinic acid (a neurotoxin) were about 2- to 2.5-fold higher than the control rats. Moreover, the brain levels in mdr1a(-/-) and mrp1(-/-) mice were 10.9- and 1.5-fold higher than those in the wild-type mice, respectively. Taken collectively, these findings indicate that PgP and Mrp1 limit the brain penetration of CTS in rodents, suggesting a possible role of PgP and MRP1 in limiting the brain penetration of CTS in patients and causing drug resistance to Danshen therapy and interactions with conventional drugs that are substrates of PgP and MRP1. Further studies are needed to explore the role of other drug transporters in restricting the brain penetration of CTS and the clinical relevance.

Topics: Alzheimer Disease; Animals; ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP-Binding Cassette Transporters; Biological Transport; Blood-Brain Barrier; Brain; Capillary Permeability; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Endothelial Cells; Infarction, Middle Cerebral Artery; Male; Mice; Mice, Knockout; Microcirculation; Multidrug Resistance-Associated Proteins; Neuroprotective Agents; Neurotoxicity Syndromes; Phenanthrenes; Plant Roots; Quinolinic Acid; Rats; Rats, Sprague-Dawley; Salvia miltiorrhiza; Stroke; Tissue Distribution; Triterpenes

The effect of triptolide, which is isolated from Tripterygium Wilfordii, on induction and development of murine AA amyloidosis was studied. In the first experiment, we examined the ability of triptolide to inhibit initiation of amyloidosis. Oral or intraperitoneal administration of 480 microg/kg/day triptolide inhibited splenic amyloid deposition in both rapid and chronic induction models of mouse AA amyloidosis. Moreover, serum amyloid A (SAA) and interleukin (IL)-6 levels were also suppressed remarkably. Triptolide also immediately decreased SAA levels and reduced the incidence of amyloidosis even under conditions of high SAA levels. In the second experiment, we evaluated the influence of triptolide on development and resorption of amyloid deposition. Amyloid deposition was induced in mice by 28 daily injections of casein. After splenic and hepatic biopsies to confirm the presence of amyloid deposits, the mice immediately started to receive daily injections of 480 microg/kg/day triptolide with or without casein. Treatment with triptolide for 35 days and 105 days prevented deposition of amyloid and promoted resorption of splenic amyloid deposits. In conclusion, we show for the first time that triptolide inhibits induction and development of experimental murine amyloidosis. These results suggest that through suppression of IL-6, triptolide can reduce production of SAA. Amyloid deposition is prevented when levels of the amyloid-forming precursor protein SAA are decreased significantly.

Topics: Amyloidosis; Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Female; Mice; Mice, Inbred ICR; Phenanthrenes; Serum Amyloid A Protein; Tripterygium

To study the preventive effect of Sodium Tanshinone II A sulfonic acid on intimal hyperplasia in rabbit iliac artery balloon injury model and explore the possible mechanism.. Thirty male pure hreed New Zealand white rabbits were undertaken experimental balloon injury in left iliac artery. Then the rabbits were assigned into treatment group (n=15) and control group (n=15), paired with weights. Sodium Tanshinone II A sulfonic acid had been injected intravenously with 7.5 - 9 mg/day for 6 days in treatment group. Saline of equivalence was given in contol group. The balloon injured arteries were harvested in the 7th, 14th, and 28th days after balloon injuy, and Paraffin sections were made. At last, HE staining, apoptosis TUNEL assay were undertaken.. (1) HE staining analysis: Media and intimal areas in treatment group at 14th day post-operation were larger than that in the 7th day (P = 0.003 and < 0.001, respectively). Media and intimal areas in treatment group decreased at the 28th day post-operation, while increased in control. Both media and intimal areas were significantly different (P < 0.001 respectively. (2) Tunel analysis discovered that, apoptosis reached peak in both treatment and control groups at the 28th post-operation. Differences of apoptosis cells counts in media and intimal between treatment and control groups were non-significant at the 7th, and 28th days, while differences at the 14th day were significant(p = 0.031 and 0.029 respectively). Apoptosis cells counting in treatment group at the 14th day increased more dramatically than that in the control.. Intravenous Sodium Tanshinone II A sulfonic acid inhibites intimal proliferation after arterial balloon injury in rabits. The effect can e partially explaineArte by the induction of apoptosis in injured artery. Clinical effect of tanshinone II A still needs further evaluation. Sodium TA-II A sulfonic acid may be of potential therapeutic value in the prevention of. To study the preventive effect of Sodium Tanshinone II A sulfonic acid on intimal by perplasia in rabbit iliac artery balloon injury model and explore the possible mechanism.. Thirty male pure breed Nexw Zealand white rabbits were un-dertaken experimental balloon injury in left iliac artery. Then the rabbits were assigned into treatment group (n=15) and control group (n=15), paired with weights. Sodium Tanshinone II A sulfonie acid had been injected intraxenously with 7.5 - 9 mg/day for 6 days in treatment group. Saline of equivalence was given in contol group. The balloon injured arteries were harvested in the 7th, 14th, and 28th days after balloon injury, and Paraffin sections were made. At last, HE staining, apoptosis TUNEL assay were undertaken.. (1) HE staining analysis: Media and intimal areas in treatment group at 14th day post-operation were larger than that in the 7th day (P = 0.003 and < 0.001, respectively). Media and intimal areas in treatment group decreased at the 28th day post-operation, while increased in control. Both media and intimal areas were significantly different (P < 0.001 respectively. (2) Tunel analysis discovered that, apoptosis reached peak in both treatment and control groups at the 28th post-operation. Differences of apoptosis cells counts in media and intimal between treatment and control groups were non-significant at the 7th, and 28th days, while differences at the 14th day were significant (p = 0.031 and 0.029 respectively). Apoptosis cells counting in treatment group at the 14th day increased more dramatically than that in the control.. Intravenous Sodium Tanshinone II A sulfonic acid inhibites intimal proliferation after arterial balloon injury in rabbits. The effect can he partially explained by the induction of apoptosis in injured artery. Clinical effect of tanshinone II A still needs further evaluation. Sodium TA-II A sulfonic acid may be of potential therapeutic value in the prevention of restenosis after angioplasty.

Topics: Abietanes; Animals; Apoptosis; Catheterization; Disease Models, Animal; Drugs, Chinese Herbal; Hyperplasia; Iliac Artery; Injections, Intravenous; Male; Phenanthrenes; Plants, Medicinal; Rabbits; Sulfonic Acids; Tunica Intima; Tunica Media

Phenanthrene imidazole 3 (MF63) has been identified as a novel potent, selective, and orally active mPGES-1 inhibitor. This new series was developed by lead optimization of a hit from an internal HTS campaign. Compound 3 is significantly more potent than the previously reported indole carboxylic acid 1 with an A549 whole cell IC(50) of 0.42 microM (50% FBS) and a human whole blood IC(50) of 1.3 microM. It exhibited a significant analgesic effect in a guinea pig hyperalgesia model when orally dosed at 30 and 100mg/kg.

Topics: Analgesics, Non-Narcotic; Animals; Disease Models, Animal; Drug Design; Guinea Pigs; Humans; Hyperalgesia; Imidazoles; Inhibitory Concentration 50; Intramolecular Oxidoreductases; Molecular Structure; Phenanthrenes; Prostaglandin-E Synthases; Rats; Structure-Activity Relationship

Hypoglycemia elicits an integrated array of CNS-mediated counterregulatory responses, including activation of the hypothalamic-pituitary-adrenal axis. The role of antecedent adrenocortical hypersecretion in impaired glucose counterregulation remains controversial. The present studies utilized the selective, nonsteroidal glucocorticoid receptor antagonist, CP-472555, as a pharmacological tool to investigate the hypothesis that hypoglycemic hypercorticosteronemia modulates CNS efferent autonomic and neuroendocrine motor responses to recurring insulin-induced hypoglycemia via glucocorticoid receptor-dependent mechanisms. Groups of adult male rats were injected s.c. with either one or four doses of the intermediate-acting insulin, Humulin neutral protamine Hagedorn (NPH), on as many days, while controls were injected with diluent alone. Animals injected with four doses of insulin were pretreated by i.c.v. administration of graded doses of the glucocorticoid receptor antagonist or vehicle alone prior to the first three doses of insulin. Repeated daily injection of NPH exacerbated hypoglycemia, attenuated patterns of glucagon and epinephrine secretion, and diminished neuronal transcriptional activation in discrete CNS metabolic loci, including the lateral hypothalamic area, dorsomedial hypothalamic nucleus, paraventricular hypothalamic nucleus, and nucleus of the solitary tract. While i.c.v. delivery of 25 or 100 ng doses of CP-472555 did not alter any of these parameters, animals treated with 500 ng exhibited circulating glucose, glucagon, and epinephrine levels that were similar to those in rats injected with one dose of insulin, as well as a reversal of recurring insulin-induced hypoglycemia-associated reductions in Fos immunolabeling in the lateral hypothalamic area, dorsomedial hypothalamic nucleus, and paraventricular hypothalamic nucleus. These results provide unique pharmacological evidence that antecedent activation of central glucocorticoid receptor is required for exacerbation of hypoglycemia during recurring insulin-induced hypoglycemia, and that these receptors mediate modulatory effects of hypoglycemic hypercorticosteronemia on autonomic efferent responses to recurring insulin-induced hypoglycemia. The data also suggest that neurons in central loci characterized here by antagonist-mediated overturn of recurring insulin-induced hypoglycemia-induced decreases in neuronal transcriptional activation may be direct or indirect substrates for this hormon

Topics: Animals; Autonomic Nervous System; Corticosterone; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Interactions; Epinephrine; Glucagon; Hypoglycemia; Hypoglycemic Agents; Hypothalamus; Insulin; Male; Neurosecretory Systems; Phenanthrenes; Proto-Oncogene Proteins c-fos; Pyridines; Rats; Rats, Sprague-Dawley; Receptors, Glucocorticoid; Transcriptional Activation

Systemic inflammatory responses contribute to mortality after thoracoabdominal aneurysm repair. Poly adenosine diphosphate (ADP) ribose polymerase (PARP) activity is known to modulate inflammation in animal models of injury. The effect of the PARP inhibitor PJ34 and genetic deletion of PARP-1(PARP -/-) on the systemic inflammatory response after thoracic aortic ischemia reperfusion (TAR) is not known.. In one group, all mice were subject to TAR followed by 48 hours of reperfusion. Treated mice (PJ, n=24) were given PJ34 IP; untreated mice (UN, n=41) received normal saline intraperitoneally. The number of mice in each group was selected to have a similar number of survivors by 48 hours. In a second group, sham animals were subjected to mediastinotomy alone (sham, n=10) without TAR, and were compared with mice with deletion of the PARP-1 isoform (PARP-1 -/-, n=11) subjected to TAR. Tissue extracts were assayed for keratinocyte derived chemokine and granulocyte colony stimulating factor. Serum was assayed for interleukin-6.. PJ34 treatment decreased mortality throughout the experimental protocol. There were no mortalities in the sham operated mice or PARP -/- mice subjected to TAR. PJ34 treatment decreased serum levels of interleukin-6 (p=0.01) and hepatic levels of interleukin-6 mRNA when compared with untreated and PARP-/- mice (p < 0.01). Only liver and kidney cytokine levels were decreased by PJ34 treatment (p < 0.05). In PARP-/- mice subjected to TAR, tissue cytokine levels were not different from those in sham mice.. PARP inhibition may represent a novel therapeutic approach to minimizing inflammatory sequelae after TAR.

Topics: Animals; Aorta, Thoracic; Cytokines; Disease Models, Animal; Male; Mice; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Reperfusion Injury; Systemic Inflammatory Response Syndrome

To test the therapeutic effect of DCB-3503, a synthetic compound derived from a natural product that inhibits NF-kappaB, on end-organ disease in the MRL-Fas(lpr) murine model of systemic lupus erythematosus (SLE).. Eight-week-old female MRL/Fas(lpr) mice were treated intraperitoneally with a low (2 mg/kg) or high (6 mg/kg) dose of DCB-3503 for 10 weeks. Control groups were administered vehicle treatment alone (negative control) or 25 mg/kg cyclophosphamide (positive control). Mice were bled before (8 weeks) and during (13 weeks) treatment, and when they were killed (20 weeks), and serum samples were analyzed for total IgM and IgG levels and autoantibody titers. When the mice were killed, spleen and lymph nodes (axillary, brachial, and cervical) were examined by flow cytometric analysis. The presence of skin and renal disease was determined by histopathologic analysis.. DCB-3503 reduced anti-double-stranded DNA and antichromatin autoantibodies and nearly abrogated inflammatory skin disease in MRL/Fas(lpr) mice; however, it had little effect on histologic kidney disease. Treated mice did not have hematologic or hepatic toxicity. These data indicate that end-organ disease in MRL/Fas(lpr) mice responds differentially to NF-kappaB inhibitor.. DCB-3503 causes significant abrogation of skin disease in MRL/Fas(lpr) mice and may potentially be beneficial in the treatment of inflammatory skin disease in SLE.

Topics: Alkaloids; Animals; Autoantibodies; Disease Models, Animal; DNA; Female; Immunoglobulin G; Immunoglobulin M; Indolizines; Lupus Erythematosus, Systemic; Lupus Nephritis; Lymphatic Diseases; Mice; Mice, Inbred MRL lpr; NF-kappa B; Phenanthrenes; Skin Diseases; Splenomegaly

The aim of this study was to investigate effects of poly(ADP-ribose) polymerase (PARP) inhibition on mesenteric vascular function and metabolism in an experimental model of cardiopulmonary bypass (CPB) with cardiac arrest. Twelve anesthetized dogs underwent 90-min hypothermic CPB. After 60 min of cardiac arrest, reperfusion was started for 40 min following application of either saline vehicle (control, n = 6) or a potent PARP inhibitor, PJ-34 (10 mg/kg iv bolus and 0.5 mg.kg(-1).min(-1) infusion for 20 min, n = 6). PJ-34 led to better recovery of cardiac output (2.2 +/- 0.1 vs. 1.8 +/- 0.2 l/min in control) and mesenteric blood flow (175 +/- 38 vs. 83 +/- 4 ml/min, P < 0.05 vs. control) after reperfusion. The impaired vasodilator response of the superior mesenteric artery to acetylcholine, assessed in the control group after CPB (-32.8 +/- 3.3 vs. -57.6 +/- 6.6% at baseline, P < 0.05), was improved by PJ-34 (-50.3 +/- 3.6 vs. -54.3 +/- 4.1% at baseline, P < 0.05 vs. control). Although plasma nitrate/nitrite concentrations were not significantly different between groups, mesenteric nitric oxide synthase activity was increased in the PJ-34 group (P < 0.05). Moreover, the treated group showed a marked attenuation of mesenteric venous plasma myeloperoxidase levels after CPB compared with the control group (75 +/- 1 vs. 135 +/- 9 ng/ml, P < 0.05). Pharmacological PARP inhibition protects against development of post-CPB mesenteric vascular dysfunction by improving hemodynamics, restoring nitric oxide production, and reducing neutrophil adhesion.

Topics: Animals; Cardiopulmonary Bypass; Disease Models, Animal; Dogs; Enzyme Inhibitors; Heart Arrest; Hemodynamics; Myocardial Reperfusion; Peroxidase; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Splanchnic Circulation

Traumatic brain injury produces peroxynitrite, a powerful oxidant which triggers DNA strand breaks, leading to the activation of poly(ADP-ribose)polymerase-1 (PARP-1). We previously demonstrated that 3-aminobenzamide, a PARP inhibitor, is neuroprotective in a model of traumatic brain injury induced by fluid percussion in rat, suggesting that PARP-1 could be a therapeutic target. In order to confirm this hypothesis, we investigated the effects of PJ34 and INO-1001, two PARP inhibitors from structural classes other than benzamide, on the post-traumatic consequences. Pre- and post-treatments with PJ34 (30 mg/kg/day) and INO-1001 (10 mg/kg/day) decrease the neurological deficit at 3 days post-injury and this deficit is still reduced at 7 days. These neurological recovery-promoting effects are associated with the inhibition of PARP-1 activation caused by trauma, as demonstrated by abolishment of immunostaining of poly(ADP-ribose). Thus, the present work strengthens strongly the concept that PARP-1 inhibition may be a suitable approach for the treatment of brain trauma.

Topics: Animals; Brain; Brain Injuries; Collagen Type XI; Disease Models, Animal; Enzyme Inhibitors; Immunohistochemistry; Indoles; Male; Nerve Degeneration; Neurons; Neuroprotective Agents; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerases; Rats; Rats, Sprague-Dawley; Recovery of Function; Solubility; Treatment Outcome

The high mortality of patients with severe acute pancreatitis (SAP) is due to multiorgan dysfunction. The mechanisms of SAP are still obscure. The aim of this study was to investigate the role of nuclear factor-kappa B (NF-kappaB) activation in rats with SAP associated with liver injury and the protection effect of triptolide against liver injury in rats with SAP.. Ninety Wistar rats were randomly divided into three groups (n=30 each group): severe acute pancreatitis (group P), treatment with triptolide (group T), and sham operation (group S). SAP models were induced by retrograde injection of 5% sodium taurocholate to the pancreatic duct. After the model was successfully established, no treatment was given to group P. In group T, triptolide (0.05 mg/ml) was injected intraperitoneally (0.2 mg/kg). In group S, the abdominal walls of rats were opened, sutured, but not treated. The rats were sacrificed after operation at 2, 6, and 12 hours, respectively. The serum levels of amylase (AMY), alanine aminotransferase (ALT), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were determined at three time points (10 rats for each time point). Liver tissues were obtained to detect the activity of NF-kappaB and to observe their pathological changes with light and electron microscopes.. The serum levels of AMY and ALT were higher in groups P and T than in group S. The serum AMY levels were significantly lower in group T than in group P at 12 hours after operation. The serum ALT levels were significantly lower in group T than in group P at 6, 12 hours after operation. At the three time points, the levels of TNF-alpha and IL-6 in groups P and T increased more significantly than in group S. In group T they were decreased more significantly than in group P at the three time points. In groups P and T, NF-kappaB activity in liver tissue increased more significantly than in group S at the three time points. The activity of NF-kappaB was higher in group P than in groups S and T at the three time points. Liver pathological damages were milder in group T than in group P under light and electron microscopes.. NF-kappaB plays an important role in the pathogenesis of liver injury in rats with SAP. Triptolide can reduce pathological damage to the liver. Its mechanism is to inhibit the activity of NF-kappaB and to decrease the release of inflammatory mediators.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Diterpenes; Epoxy Compounds; Female; Immunohistochemistry; Interleukin-6; Liver; Liver Diseases; Male; NF-kappa B; Pancreatitis; Phenanthrenes; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

The putative selective estrogen receptor modulator (+)-Z-bisdehydrodoisynolic acid (Z-BDDA) has been found to improve cardiovascular risk in rodents. The objective of this study was to investigate the effectiveness of (+)-Z-BDDA compared with the antidiabetic drug, rosiglitazone, in treating obesity and risk factors associated with the metabolic syndrome.. Female Zucker Diabetic Fatty rats were randomly assigned to three treatment groups for 29 weeks: control (C), 1.8 mg (+)-Z-BDDA/kg diet [control diet + (+)-Z-BDDA (CB)], or 100 mg rosiglitazone/kg diet [control diet + rosiglitazone (CR)]. At sacrifice, physiological, biochemical, and molecular parameters were examined.. CB animals gained less weight and exhibited a decrease in total body lipids (p < 0.05) as compared with C or CR rats. Body weight and total body lipids were the highest in CR rats (p < 0.05). Liver weights in CB and CR rats were lower (p < 0.05) than in C rats, whereas kidney weights were lower in CB (p < 0.05) than in C and CR animals. Fasting plasma glucose was lower (p < 0.05) in the CB and CR animals when compared with C animals. C rats exhibited the highest concentration of total plasma cholesterol, and CR-treated rats exhibited the lowest concentration. Plasma triglycerides followed the same pattern as plasma cholesterol. Histomorphometry of heart vasculature revealed that CB and CR treatments produced a significant shift from small to large venules and arterioles compared with C (p < 0.05). Liver expression profiles of peroxisome proliferator-activated receptor (PPAR) alpha, PPARgamma, and PPAR-regulated genes revealed encouraging CB-induced effects.. These results suggest that (+)-Z-BDDA may have applications in treating obesity and complications associated with the metabolic syndrome.

Topics: Animals; Blood Glucose; Body Weight; Cholesterol; Coronary Vessels; Diabetes Mellitus, Experimental; Disease Models, Animal; Female; Gene Expression; Hypoglycemic Agents; Kidney; Liver; Metabolic Syndrome; Obesity; Organ Size; Phenanthrenes; PPAR alpha; PPAR gamma; Random Allocation; Rats; Rats, Zucker; Risk Factors; Rosiglitazone; Selective Estrogen Receptor Modulators; Thiazolidinediones; Triglycerides

To observe change of iliac artery SMC collagen fiber after balloon injury in tanshinone II A treatment group.. Thirty male pure breed New Zealand white rabbits were underwent experimental balloon injury in left iliac artery. Then the rabbits were assigned into treatment group (n = 15) and control group (n = 15). Tanshinone II A sulfonic acid natrium was injected intravenously in treatment group. Saline was venously injected in control group. The balloon injured arteries were harvested in the 7th, 14th, and 28th days after surgery.. Analysis of Collagen fibers staining found that a weak positive staining was shown at the 7th day post-operation in both groups,while a strong positive result was seen at the 14th day in control group. Positive results were even shown at the 14th day in treatment group,but less strong than that of the control. At the 28th day post-operation, collagen fiber staining become weak in treatment group, but increased in the control. Difference between both group in the 28th day post-operation was significant (P = 0.017). Time-dependence tendency of collagen fiber changing was statistically significant (P = 0.005).. Tanshinone II A suppress stack of extracellular matrix after balloon injury so as to prevent restenosis.

Topics: Abietanes; Animals; Catheterization; Collagen; Coronary Restenosis; Disease Models, Animal; Iliac Artery; Male; Muscle, Smooth, Vascular; Phenanthrenes; Plants, Medicinal; Rabbits; Salvia; Tunica Intima; Tunica Media

Poly(ADP-ribose) polymerase (PARP) activity has been implicated in the pathogenesis of several central nervous system (CNS) disorders. For example, the presence of extensive poly(ADP)ribosylation in CNS tissues from animals with experimental allergic encephalomyelitis (EAE) indicates that PARP activity may be involved in this inflammatory disease process. Using PJ34 [N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N, N-dimethylacetamide.HCl], a selective PARP inhibitor, we studied the mechanisms through which PARP activity may contribute to the onset of acute EAE. PLSJL mice immunized with myelin antigens were treated with PJ34, and the effects on the progression of EAE and several other parameters relevant to the disease process were assessed. PJ34 exerted therapeutic effects at the onset of EAE that were associated with reduced CNS inflammation and the maintenance of neurovascular integrity. Expression of genes encoding the intercellular adhesion molecule-1 (ICAM-1) and the inflammatory mediators interferon-gamma, tumor necrosis factor-alpha, and inducible nitric-oxide synthase were decreased in CNS tissues from drug-treated animals. Administration of PJ34 biased the class of myelin basic protein (MBP)-specific antibodies elicited from IgG2a to IgG1 and IgG2b and modulated antigen-specific T-cell reactivity. Therefore, the mode of action of PJ34 at the onset of EAE is likely mediated by a shift in the MBP-specific immune response from a proinflammatory Th1 toward an anti-inflammatory Th2 phenotype.

Topics: Adjuvants, Immunologic; Animals; Blood-Brain Barrier; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Intercellular Adhesion Molecule-1; Mice; Myelin Basic Protein; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Spinal Cord Diseases; T-Lymphocytes; Th2 Cells; Tumor Necrosis Factor-alpha

Overactivation of the DNA repair enzyme poly(ADP-ribose) polymerase (PARP) contributes to ischemic brain injury. Because PARP upregulates proinflammatory genes, we investigated whether inducible nitric oxide synthase (iNOS), a gene involved in the deleterious effects of postischemic inflammation, participates in the mechanisms by which PARP activation contributes to cerebral ischemic injury.. The middle cerebral artery (MCA) was occluded in mice for 20 minutes using an intravascular filament, and injury volume was measured 72 hours later in Nissl-stained brain sections. mRNA expression was assessed in the postischemic brain by the quantitative "real-time" polymerase chain reaction.. The PARP inhibitor PJ34 reduced infarct volume and attenuated postischemic iNOS mRNA upregulation by 72%. To determine whether iNOS contributes to the toxicity of PARP, the iNOS inhibitor aminoguanidine was co-administered with PARP inhibitors. Unexpectedly, co-administration of PARP and iNOS inhibitors, or treatment of iNOS-null mice with PARP inhibitors, abrogated the protective effect afforded by iNOS or PARP inhibition alone. The loss of neuroprotection was associated with upregulation of the inflammatory genes iNOS, intercellular adhesion molecule-1, and gp91(phox).. The results suggest that iNOS expression contributes to the deleterious effects exerted by PARP activation in cerebral ischemia. However, iNOS activity is required for the protective effect of PARP inhibition and, conversely, PARP activity must be present for iNOS inhibition to be effective. The findings unveil a previously unrecognized deleterious interaction between iNOS and PARP that is relevant to the development of combination therapies for ischemic stroke.

Topics: Animals; Brain Ischemia; Disease Models, Animal; Guanidines; Infarction, Middle Cerebral Artery; Intercellular Adhesion Molecule-1; Male; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; NADPH Oxidase 2; NADPH Oxidases; Neuroprotective Agents; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Polymerase Chain Reaction; RNA, Messenger; Up-Regulation

Excessive activation of poly(ADP-ribose) polymerase-1 (PARP-1), a nuclear enzyme catalyzing the transfer of ADP-ribose units from NAD to acceptor proteins, induces cellular energy failure by NAD and ATP depletion and has been proposed to play a causative role in a number of pathological conditions, including ischemia/reperfusion injury. In this study, we used an in vitro enzyme activity assay to characterize a series of newly synthesized isoquinolinone derivatives as potential PARP-1 inhibitors. Several compounds displayed powerful inhibitory activity: thieno[2,3-c]isoquinolin-5-one (TIQ-A) displayed a submicromolar IC50 of 0.45 +/- 0.1 microM, whereas the 5-hydroxy and 5-methoxy TIQ-A derivatives had IC50 values of 0.39 +/- 0.19 and 0.21 +/- 0.12 microM, respectively. We then examined the neuroprotective effects of the newly characterized compounds in cultured mouse cortical cells exposed to 60 min of oxygen and glucose deprivation (OGD). When PARP-1 inhibitors were present in the incubation medium during OGD and the subsequent 24-h recovery period, they significantly attenuated neuronal injury. TIQ-A provided neuroprotection even when added to the culture 30 min after OGD and was able to reduce the early activation of PARP induced by OGD as detected by flow cytometry. When the IC50 values observed in the PARP-1 activity assay for selected compounds were compared with their IC50 values for the neuroprotective activity, a significant correlation (r = 0.93, P < 0.01) was observed. Our results suggest that TIQ-A and its derivatives are a new class of neuroprotectants that may be helpful in studies aimed at understanding the involvement of PARP-1 in physiology and pathology.

Topics: Animals; Brain Ischemia; Cells, Cultured; Disease Models, Animal; Isoquinolines; Mice; Neuroprotective Agents; Phenanthrenes; Piperidines; Poly(ADP-ribose) Polymerase Inhibitors; Thiophenes

Activation of the poly(ADP-ribose)polymerase (PARP), a highly energy-consuming DNA-repairing enzyme, plays a crucial role in the pathogenesis of multiorgan failure. Most results, however, were derived from experiments with hypodynamic shock states characterized by a markedly decreased cardiac output (CO) and/or using a pretreatment approach. Therefore, we investigated the effects of the novel potent and selective PARP-1 inhibitor PJ34 in a posttreatment model of long-term, volume-resuscitated porcine endotoxemia. Anesthetized, mechanically ventilated and instrumented pigs received continuous intravenous (i.v.) lipopolysaccharide (LPS) over 24 h. Hydroxyethyl starch was administered to maintain a mean arterial pressure > 65 mmHg. After 12 h of LPS infusion, the animals were randomized to receive either vehicle (Control, n = 9) or i.v. PJ34 (n = 6; 10 mg/kg over 1 h followed by 2 mg/kg/h until the end of the experiment). Measurements were performed before as well as at 12, 18, and 24 h of LPS infusion. In all animals CO increased because of reduced systemic vascular resistance (SVR) and fluid resuscitation. PJ34 further raised CO (P < 0.05 vs. control group) as the result of a higher stroke volume indicating its positive inotropic effect. In addition, it diminished the rise in the ileal mucosal-arterial PCO2 gap, which returned to baseline levels at 24 h of LPS, and improved the gut lactate balance (P = 0.093 PJ34 vs. control) together with significantly lower portal venous lactate/pyruvate ratios. By contrast, it failed to influence the LPS-induced derangements of liver metabolism. Incomplete PARP inhibition because of dilutional effects and/or an only partial efficacy when used in post-treatment approaches may account for this finding.

Topics: Acid-Base Equilibrium; Animals; Disease Models, Animal; Endotoxemia; Enzyme Inhibitors; Female; Hemodynamics; Lactates; Lipopolysaccharides; Liver; Liver Circulation; Male; Oxygen; Oxygen Consumption; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Reference Values; Splanchnic Circulation; Swine

The effect of a chemical mixture on the dermal penetration of arsenic or nickel was assessed by applying arsenic-73 or nickel-63 alone or with the chemical mixture to dermatomed male pig skin samples in flow-through diffusion cells. The chemical mixture consisted of chloroform, phenanthrene, and toluene for arsenic penetration studies and phenol, toluene, and trichloroethylene (TCE) for nickel studies. These are predominant chemicals found at hazardous waste sites. Arsenic and nickel bind to skin after dermal exposure. Total penetration of arsenic and nickel in the chemical mixture were significantly increased by 33% and 20% compared to arsenic and nickel alone, respectively. While more radioactivity penetrated skin with chemical treatment than metal alone, significantly less radioactivity was loosely adsorbed to skin and could be easily washed off from the skin surface with soap and water. The results of this study indicate that the potential health risk from dermal exposure to arsenic or nickel is enhanced if other chemicals are present.

Topics: Animals; Arsenic Poisoning; Arsenicals; Chloroform; Disease Models, Animal; Drug Synergism; Hazardous Waste; Male; Nickel; Phenanthrenes; Skin Absorption; Swine; Time Factors; Toluene

Early life stages of rainbow trout were exposed to different regimes of water-borne retene (7-isopropyl-1-methylphenanthrene) to determine if there is an ontogenic stage particularly sensitive to retene toxicity, and if cytochrome P-4501A (CYP1A) induction is a forerunner to blue sac disease (BSD), the syndrome of toxicity. CYP1A protein concentrations, measured by immunohistochemistry, were first detected during organogenesis, when organ and enzyme systems are first being developed, and steadily increased until swim-up. The prevalence of signs of BSD rose 1 wk following a marked increase in CYP1A activity after hatch, suggesting that CYP1A induction is related to BSD. The larval stage was the most sensitive to retene toxicity, based on CYP1A induction and a high prevalence of BSD. The most common signs of BSD were hemorrhaging, yolk-sac edema, and mortality, but hemorrhaging was the first and most frequently observed response. Tissue concentrations of retene were elevated just after fertilization, but decreased steadily as fish developed to the swim-up stage, most likely due to the establishment of more efficient metabolic and excretory systems in later stages of development.

Topics: Animals; Biomarkers; Chorion; Cytochrome P-450 CYP1A1; Disease Models, Animal; Environmental Exposure; Environmental Monitoring; Fetal Diseases; Fish Diseases; Fishes; Geologic Sediments; Immunohistochemistry; Larva; Neurotoxicity Syndromes; Oncorhynchus mykiss; Organogenesis; Phenanthrenes; Prevalence; Tissue Distribution; Water Pollutants, Chemical

We investigated the suppressive effect of triptolide (TRD), a purified component from a traditional Chinese herb, Tripterygium wilfordii Hook F. (TWHf), on uveitogenic peptide (K2)-induced experimental autoimmune uveoretinitis (EAU). K2-peptide immunized B10.A mice were divided into four groups. One group was EAU control which was treated with PBS. The other two groups were treated with TRD with different time courses (from day 0 to day 28 and from day 14 to day 28). The last group was treated with Cyclosporin A (CsA) as a positive control of the treatment. TRD was administered at dose of 0.1 mg/kg/day (i.p.). CsA was administered at dose of 20 mg/kg/day (i.p.) from day 0 to day 28 during whole period of EAU induction. The data showed that the EAU was suppressed in the whole period of TRD-treated mice, but was not in TRD-treated mice from day 14 to day 28 following immunization. The inhibition of EAU induced by TRD treatment was comparable to CsA-treated mice. The K2-specific lymphocyte proliferation and mRNA expressions of Th1-type cytokines (IL-12p40, IFN-gamma and TNF-alpha) in draining lymph node and inflamed eyes were reduced in TRD-treated mice. The K2-specific IFN-gamma production in the draining lymph node cells (LNC) of TRD-treated mice (whole period) was significantly inhibited. This effect was not related to an apoptotic effect of TRD on CD4+ T cells. Our results suggested that TRD suppressed the induction of EAU by down-regulating Th1-type response in B10.A mice. This preventive effect on EAU induction may be related to the inhibition of TRD on T cell priming and activation.

Topics: Animals; Autoimmune Diseases; Cytokines; Disease Models, Animal; Diterpenes; Down-Regulation; Epoxy Compounds; Female; Immunosuppressive Agents; Injections, Intraperitoneal; Mice; Mice, Inbred Strains; Phenanthrenes; Retinitis; Th1 Cells; Uveitis

We have investigated the role of the activation of nuclear poly(ADP-ribose) polymerase (PARP) enzyme, a mediator of downstream nitric oxide toxicity, using a combined approach of pharmacological inhibition and genetic disruption in a ligature-induced-periodontitis model in rats and mice. Immunohistochemical analysis revealed significantly increased poly(ADP-ribose) nuclear staining (indicative of PARP activation) in the subepithelial connective tissue of the ligated side compared with the non-ligated side. Ligation-induced periodontitis resulted in marked plasma extravasation in the gingivomucosal tissue and led to alveolar bone destruction compared with the non-ligated side, as measured by the Evans blue technique and by videomicroscopy, respectively. PARP inhibition with PJ34, as well as genetic PARP-1 deficiency, significantly reduced the extravasation and the alveolar bone resorption of the ligated side compared with controls. Thus, PARP activation contributes to the development of periodontal injury. Inhibition of PARP may represent a novel host response modulatory approach for the therapy of periodontitis.

Topics: Alveolar Bone Loss; Animals; Cell Nucleus; Connective Tissue; Disease Models, Animal; Enzyme Activation; Gingiva; Ligation; Male; Mice; Mice, Knockout; Mouth Mucosa; Nuclear Proteins; Periodontitis; Phenanthrenes; Plasma; Poly Adenosine Diphosphate Ribose; Rats; Rats, Wistar

Nonterpenoid and diterpenoid phenanthrenequinones (PACs) have been found in the plant kingdom. Some of them occur in plants used in traditional Chinese medicine like Tan-Shen whereas others are constituents of orchids that are popular as ornamental plants.. Case reports and our own observations in orchid nurseries suggest that some or even all of these PACs possess a distinct sensitizing potency. Occasional exposure (particularly of botanists) to field-grown orchids, as well as occupational contact with sawdust of PAC-containing tropical timbers, caused allergic contact dermatitis. However, experimental studies in guinea pigs to determine the sensitizing capacity of PACs have not been performed so far.. Guinea pigs were sensitizied by a modified Freund's complete adjuvant method with four naturally occurring and 22 synthetic PACs in order to find out which and how many substituents at the carbons of the three rings of the PAC will influence the sensitizing power of the molecule. Subsequently, the lowest unoccupied molecular orbital (LUMO) coefficients were calculated to show whether a correlation exists between chemical reactivity and sensitizing capacity.. Sensitizing capacity was found to be strong in two PACs, moderate in eight PACs, and weak in ten PACs. Five PACs were extremely weak in sensitizing capacity, and one PAC was completely negative. Two substituents on the left-hand carbons C-7 and C-8 of ring C were shown to be responsible for a strong sensitizing capacity. One methoxy group alone or three of them, especially when localized at C-5, decreased the sensitizing capacity to moderate. Substitution with a methoxy group at C-3 and/or at C-2 of the quinonoid ring itself (ring A) led to a weak sensitizing capacity. The ortho-quinones 1,2-PAC and 9,10-PAC were also weakly sensitizing. In fact, LUMO coefficient calculations corroborated a good correlation between chemical reactivity and sensitizing capacity.. Substitution with methoxy groups at C-7 and/or at C-8 of ring C of 1,4-phenanthrenequinone increases the LUMO coefficients at the 2,3 double bond of ring A and thus facilitates nucleophilic substitution of protein nitrogen or sulfur nucleophiles at this electron-deficient double bond. The four naturally occurring PACs that were investigated--cypripedin, denbinobin, annoquinone-A, and latinone--do not fulfill these criteria and are thus only weak sensitizers. However, as-yet-unstudied phenanthrenequinones occurring in plants or trees and having no substituents at C-2 or C-3 of the quinonoid ring must be considered potentially strong allergens.

Topics: Allergens; Animals; Dermatitis, Allergic Contact; Disease Models, Animal; Guinea Pigs; Patch Tests; Phenanthrenes; Structure-Activity Relationship

We recently reported that (Lamiaceae) may alleviate CCl(4)-induced acute hepatotoxicity in rats, possibly blocking the formation of free radicals generated during CCl(4) metabolism. Carnosol, one of the main constituents of Rosmarinus, has been shown to have antioxidant and scavenging activities. Therefore, it is plausible to expect that carnosol may mediate some of the effects of Rosmarinus on oxidative stress consequences induced by CCl(4) in the liver.. We evaluated the effectiveness of carnosol to normalize biochemical and histological parameters of CCl(4)-induced acute liver injury.. Male Sprague Dawley rats (n = 5) injured by CCl(4) (oral dose 4 g/kg of body weight) were treated with a single intraperitoneal dose (5 mg/kg) of carnosol. Twenty-four hours later, the rats were anaesthetized deeply to obtain the liver and blood, and biochemical and histological parameters of liver injury were evaluated.. Carnosol normalized bilirubin plasma levels, reduced malondialdehyde (MDA) content in the liver by 69%, reduced alanine aminotransferase (ALT) activity in plasma by 50%, and partially prevented the fall of liver glycogen content and distortion of the liver parenchyma.. Carnosol prevents acute liver damage, possibly by improving the structural integrity of the hepatocytes. To achieve this, carnosol could scavenge free radicals induced by CCl(4), consequently avoiding the propagation of lipid peroxides. It is suggested that at least some of the beneficial properties of Rosmarinus officinalis are due to carnosol.

Topics: Abietanes; Acute Disease; Animals; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Hepatocytes; Liver; Liver Diseases; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley

Reactive oxygen and nitrogen species are overproduced in the cardiovascular system during circulatory shock. Oxidant-induced cell injury involves the activation of poly(ADP-ribose) polymerase (PARP). Using a dual approach of PARP-1 suppression, by genetic deletion or pharmacological inhibition with the new potent phenanthridinone PARP inhibitor PJ34 [the hydrochloride salt of N-(oxo-5,6-dihydro-phenanthridin-2-yl)-N,N-dimethylacetamide], we studied whether the impaired cardiac function in endotoxic shock is dependent upon the PARP pathway. Escherichia coli endotoxin (lipopolysaccharide, LPS) at 55 mg/kg, i.p., induced a severe depression of the systolic and diastolic contractile function, tachycardia, and a reduction in mean arterial blood pressure in both rats and mice. Treatment with PJ34 significantly improved cardiac function and increased the survival of rodents. In addition, LPS-induced depression of left ventricular performance was significantly less pronounced in PARP-1 knockout mice (PARP(-/-)) as compared with their wild-type littermates (PARP(+/+)). Thus, PARP activation in the cardiovascular system is an important contributory factor to the cardiac collapse and death associated with endotoxin shock.

Topics: Animals; Death; Disease Models, Animal; Drug Interactions; Endotoxins; Enzyme Activation; Gene Deletion; Heart Diseases; Heart Function Tests; Lipopolysaccharides; Male; Mice; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Rats; Rats, Wistar

1. This study was designed to compare the proarrhythmic activity of the antimalarial drug, halofantrine and the antihistamine, terfenadine, with that of clofilium a K(+) channel blocking drug that can induce torsade de pointes. 2. Experiments were performed in pentobarbitone-anaesthetized, open-chest rabbits. Each rabbit received intermittent, rising dose i.v. infusions of the alpha-adrenoceptor agonist phenylephrine. During these infusions rabbits also received increasing i.v. doses of clofilium (20, 60 and 200 nmol kg(-1) min(-1)), terfenadine (75, 250 and 750 nmol kg(-1) min(-1)), halofantrine (6, 20 and 60 micromol kg(-1)) or vehicle. 3. Clofilium and halofantrine caused dose-dependent increases in the rate-corrected QT interval (QTc), whereas terfenadine prolonged PR and QRS intervals rather than prolonging cardiac repolarization. Progressive bradycardia occurred in all groups. After administration of the highest dose of each drug halofantrine caused a modest decrease in blood pressure, but terfenadine had profound hypotensive effects resulting in death of most rabbits. 4. The total number of ventricular premature beats was highest in the clofilium group. Torsade de pointes occurred in 6 out of 8 clofilium-treated rabbits and 4 out of 6 of those which received halofantrine, but was not seen in any of the seven terfenadine-treated rabbits. 5. These results show that, like clofilium, halofantrine can cause torsade de pointes in a modified anaesthetized rabbit model whereas the primary adverse effect of terfenadine was cardiac contractile failure.

Topics: Animals; Antimalarials; Disease Models, Animal; Electrocardiography; Histamine Antagonists; Male; Phenanthrenes; Potassium Channel Blockers; Quaternary Ammonium Compounds; Rabbits; Terfenadine; Torsades de Pointes

To determine whether activation of the nuclear enzyme poly(adenosine 5'-diphosphate [ADP]-ribose) synthetase (PARS) contributes to mortality rate, myocardial dysfunction, and cardiovascular collapse in a porcine model of sepsis induced by implantation of an infected clot.. Prospective, random animal study.. Research laboratory at Rush Presbyterian St. Luke's Medical Center.. Twenty pigs were chronically instrumented with intracardiac transducers to measure left ventricular pressure, sonomicrometer crystals in the left ventricle to measure short axis diameter, an ultrasonic flow meter to measure cardiac output, and catheters in the pulmonary artery and aorta to measure blood pressures and collect samples.. By using a randomized study design, we administered either the novel potent PARS inhibitor PJ34 (10 mg/kg for 1 hr, 2 mg x kg(-1) x hr(-1) for 96 hrs) or vehicle to pigs immediately before intraperitoneal implantation of Escherichia coli 0111.B4 (2.3 +/- 0.1 x 10(10) colony-forming units/kg)-laden fibrin clots to produce peritonitis and bacteremia.. In vehicle-treated pigs, 12% survival was recorded at 24 hrs, whereas 83% and 66% survival was recorded in the PJ34-treated animals at 24 and 96 hrs, respectively (p <.05). PJ34 treatment attenuated bacteremia-induced increases in systemic and pulmonary vascular resistances. In controls, peritonitis induced rapid increase in plasma tumor necrosis factor-alpha. PJ34 treatment significantly attenuated this cytokine response. The formation of peroxynitrite and the activation of PARS were confirmed in hearts and lungs of the septic pigs by the immunohistochemical detection of nitrotyrosine and poly(ADP-ribose), respectively. Inhibition of PARS with PJ34 abolished poly(ADP-ribose) formation in septic animals.. Treatment with a potent PARS inhibitor improved survival and cardiovascular status and attenuated an important mediator component of the inflammatory response in a lethal porcine model of sepsis.

Topics: Animals; Bacterial Infections; Disease Models, Animal; Enzyme Inhibitors; Escherichia coli Infections; Hemodynamics; Peritonitis; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Prospective Studies; Random Allocation; Swine; Tumor Necrosis Factor-alpha

Topics: Animals; Asthma; Caspases; Disease Models, Animal; Diterpenes; Epoxy Compounds; Guinea Pigs; Interleukin-1 Receptor-Associated Kinases; Lung; Male; Phenanthrenes; Proto-Oncogene Proteins c-bcl-2; Tripterygium

Focal cerebral ischemia activates the nuclear protein poly(ADP-ribose) polymerase (PARP) by single DNA strand breaks which leads to energy depletion and cell necrosis. Deletion or inhibition of PARP protects against ischemic brain injury. Here we examined the neuroprotective effect of PJ34, a novel potent inhibitor of PARP in vitro and in vivo. Serum-free primary neuronal cultures derived from rat cortex (E15-17) and kept in culture for 10 days were exposed to oxygen glucose deprivation (OGD) in vitro. Neuronal injury was quantified by LDH release after 24 h. Pretreatment with 30-1000 nM PJ34 significantly protected from OGD-induced cell injury in a dose-dependent manner. For in vivo experiments SV/129 mice were treated with PJ34 (50 microg) by intraperitoneal injection 2 h before 1 h middle cerebral artery occlusion (MCAo) and again 6 h later. Twenty-three h after reperfusion ischemic injury was significantly decreased compared to vehicle-treated controls (infarct volume reduction of 40%, p<0.05). Similarly, in a rat model of MCAo (2 h occlusion followed by up to 22 h reperfusion), PJ34 administration (10 mg/kg i.v.) significantly reduced infarct size, and the effect of the drug was maintained even if it was given as late as 10 min prior to reperfusion time. PJ34 significantly protected in a 4 h, but not in a 24 h permanent occlusion model. In conclusion, PJ34, a novel, potent inhibitor of PARP exerts massive neuroprotective agents, with a significant therapeutic window of opportunity. The present work strengthens the concept that pharmacological PARP inhibition may be a suitable approach for the treatment of acute stroke in man.

Topics: Animals; Brain; Brain Ischemia; Cells, Cultured; Culture Media, Serum-Free; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme Inhibitors; Infarction, Middle Cerebral Artery; Mice; Neurons; Neuroprotective Agents; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Rats; Rats, Wistar; Stroke

Salvia miltiorrhiza is a traditional Chinese medicine which has been well documented for its anti-cancer effects. Based on the structure of danshinone, one of the active compounds derived from Salvia miltiorrhiza, we synthesized a simplified phenolic analog, S-3-1, and tried to explore its possible actions in preventing the development of cancer. With the Ames test, S-3-1 was found to efficiently suppress the mutagenicity of benzo[alpha]pyrene. This result is consistent with the inhibitory effect of S-3-1 on the activation of benzo[alpha]pyrene by hepatic microsomal enzymes. Besides the anti-initiation effects, S-3-1 could significantly inhibit the croton oil-induced increase of mouse skin epithermal ornithine decarboxylase activity. Moreover, S-3-1 quenched both superoxide and hydroxyl free radicals whereas it inhibited lipid peroxidation in the in vitro model. These results suggest that S-3-1 might act as anti-initiation and anti-promotion agents through reversing the biochemical alterations induced by carcinogen during carcinogenesis. Therefore, we further investigated the effects of S-3-1 on carcinogenesis. In vitro, S-3-1 inhibited the benzo[alpha]pyrene-induced transformation of V79 Chinese hamster lung fibroblasts. At 10-40 mg/kg, S-3-1 was found to inhibit the development of DMBA/croton oil-induced skin papilloma in mice through decreasing the incidence of papilloma, prolonging the latent period of tumor occurrence and reducing tumor number per mouse in a dose-dependent manner. We concluded from this study that S-3-1 might be developed as a new chemopreventive drug.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Anticarcinogenic Agents; Benzo(a)pyrene; Benzofurans; Bepridil; Biphenyl Compounds; Cell Transformation, Neoplastic; Cells, Cultured; Cricetinae; Croton Oil; Cysteine; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Epithelial Cells; Fibroblasts; Free Radical Scavengers; Hypoxanthine; In Vitro Techniques; Iron; Lipid Peroxidation; Lung; Male; Medicine, Chinese Traditional; Mice; Mice, Inbred ICR; Microsomes, Liver; Molecular Structure; Mutagens; Ornithine; Ornithine Decarboxylase; Papilloma; Pentetic Acid; Phenanthrenes; Picrates; Plants, Medicinal; Rats; Salmonella; Skin; Skin Neoplasms; Spectrometry, Mass, Electrospray Ionization; Structure-Activity Relationship; Xanthine Oxidase

Chinese herbs nephropathy (CHN) is a new type of subacute interstitial nephritis that is attributed to aristolochic acid (AA), which inadvertently has been included in slimming pills. The contribution of other simultaneously prescribed drugs remains disputed. In the present study, the effects of a chronic intake of AA given as a single drug was evaluated through renal histology and function in rabbits.. Female New Zealand White rabbits were injected intraperitoneally with either 0.1 mg AA/kg or with saline 5 days a week for 17 to 21 months. Body weight, renal function, and urinary excretion of glucose and low molecular weight proteins were monitored prior to sacrifice at the end of the study period.. All animals given AA developed renal hypocellular interstitial fibrosis, which was classified into three patterns. Fibrosis was confined to medullary rays (MRs) in pattern I (N = 3), extended to the outer cortical labyrinth (OCL) in pattern II (N = 2), and eventually to the inner cortical labyrinth (ICL) in pattern III (N = 6). Fibrosis in MR and OCL was associated with mainly proximal tubular epithelial cell flattening. All treated animals displayed urothelial atypia. Three of them also developed tumors of the urinary tract. No significant pathologic changes were found in control rabbits. AA-treated animals differed from controls by an impaired growth, increased serum creatinine, glucosuria, tubular proteinuria, and anemia.. The observed pattern of renal histopathological lesions and disorders of the renal function, as well as urothelial atypia and malignancy, are very reminiscent of CHN. Our observations therefore support a causal role of AA alone in the genesis of this new nephropathy.

Topics: Animals; Aristolochic Acids; Disease Models, Animal; Drugs, Chinese Herbal; Enzyme Inhibitors; Female; Fibrosis; Kidney; Nephritis, Interstitial; Organ Size; Phenanthrenes; Rabbits; Stomach

Tylophora alkaloids have been shown to have antiasthmatic, antiinflammatory and antianaphylactic properties. Since all these disorders are a consequence of altered immunological status, the effect of these alkaloids on model immune reactions were studied. Crude extract of the leaves of Tylophora indica inhibited delayed hypersensitivity reaction to sheep red blood cells in rats when the alkaloid mixture was administered before and after immunization with these cells. The alkaloid mixture also inhibited contact sensitivity to dinitro-fluorobenzene in mice when given prior to or after contact sensitization. Lymphocytes taken from contact sensitized mice, when treated with tylophora alkaloid in vitro and transferred into naive syngeneic hosts, could suppress the transfer of delayed type hypersensitivity (DTH) response. However, the tylophora alkaloids could not suppress primary humoral (IgM) immune response to SRBC in mice at the same dose. These studies suggest that tylophora alkaloids suppress cellular immune responses when administered at any stage during the immune response.

Topics: Alkaloids; Animals; Apocynaceae; Cell Survival; Dinitrofluorobenzene; Disease Models, Animal; Dose-Response Relationship, Drug; Erythrocytes; Hypersensitivity, Delayed; Immunity, Cellular; Indolizines; Isoquinolines; Lymphocytes; Mice; Phenanthrenes; Phytotherapy; Plant Extracts; Rats; Rats, Wistar; Sheep

To study chronic renal interstitial fibrosis induced by aristolochic acid (AA) in animal models.. Female Wistar rats were divided into two groups: AA group (n = 42) peritoneally injected with AA (5 mg.kg-1.d-1) for 16 weeks and control group (n = 5) peritoneally injected with normal saline (2 ml/d) for 16 weeks. The body weight of rats was taken. At week 8, 12, 16, and 24 six rats were killed in the AA group. The five rats in the control group were killed at week 24. Specimens of lood and urine were taken before the animals were killed. Specimens of renal tissue were taken after the animals were killed. Twenty-four hour urine protein, urine beta 2 microglobulin (beta 2-MG), and renal function were tested regularly. Pathological examination, including tubulo-interstitial area calculation, was made to the renal specimens.. The body weight of rats in AA group became significantly lower than that of the control rats after 16 weeks' AA injection (P < 0.01). The blood urea nitrogen (BUN) and serum creatinine (Scr) in AA group increased significantly than those of the control group at the 16th, 20th, and 24th weeks (P < 0.05). Optical microscopy showed tubular-interstitial damage in AA group in 16 weeks. Renal tubular atrophy and multifocal renal interstitial fibrosis were shown in 24 weeks. The area of renal tubule increased and the area of the lumen remarkably decreased at week 16 compared with those in the control group. The area of renal tubule decreased remarkably and the area of renal interstitial greatly in the AA group at week 24. Electrical microscopy showed increase of primary and secondary lysosomes and diasappearence of part of brush border of tubular endothelial cells at week 16, and accumulation of secondary lysosome in cytoplasm at weeks 20 and 24 in the AA group. Histoimmunofluorescence showed that IgG, IgA, IgM, C3, and C1q were negative in the renal tissue of experimental animals.. Animal models with chronic renal tubuloointerstitial nephropathy induced by aristolochic acid were successfully established. Aristolochic acid may have chronic toxicity to renal tissues, and cause chronic renal interstitial fibrosis in rats.

Topics: Animals; Aristolochic Acids; Blood Urea Nitrogen; Body Weight; Carcinogens; Chronic Disease; Creatine; Disease Models, Animal; Female; Fibrosis; Kidney Tubules, Proximal; Nephritis, Interstitial; Phenanthrenes; Rats; Rats, Wistar

The declining efficacy of antimalarial drugs against Plasmodium falciparum strains has been reported from several endemic regions of the world. Strategic evaluation of several pharmacological agents in combination with chloroquine to enhance the sensitivity of the latter against resistant parasites has been documented in several studies. However no attempts have been directed to monitor the efficacy of such biological response modifiers for reversing the resistance to halofantrine. In the present study the comparative efficacy of a total of 22 pharmacological agents representing diverse categories including antihistamines, antidepressants, calcium channel blockers, neuroleptics etc. has been determined in combination with halofantrine against halofantrine resistant Plasmodium yoelii nigeriensis. Results show significant potentiation of the efficacy of halofantrine when administered concurrently with histamine H1 receptor antagonists cyproheptadine and ketotifen. Combination with pheniramine, amitriptyline, verapamil and penfluridol also produced moderate degree of potentiation which was well marked during the early phase of progression of parasitaemia.

Topics: Animals; Antimalarials; Cyproheptadine; Disease Models, Animal; Drug Resistance; Female; Male; Mice; Phenanthrenes; Plasmodium falciparum

This paper reports the effect of triptolide (a diterpenoid triepoxide) on the development of monocrotaline (MCT)-induced pulmonary hypertension in pneumonectomized rats. Male Sprague- Dawley rats were injected with MCT (60 mg/kg) on Day 7 after left pneumonectomy. Rats received therapy from Day 5 to 35 with triptolide (0.25 mg/kg intraperitoneally, every other day, n = 10), or vehicle (0.1 ml of ethanol/cremophor intraperitoneally, every other day, n = 10). By Day 35, triptolide-treated rats demonstrated lower mean pulmonary arterial pressure (mPAP) than vehicle-treated rats (mPAP 21 +/- 3 versus 42 +/- 5 mm Hg, p < 0.001). Triptolide-treated rats also had significantly less right ventricular hypertrophy (RVH) and pulmonary arterial neointimal formation. In a rescue experiment, rats initiated therapy on Day 21. At Day 35, vehicle-treated rats (n = 4) had higher mPAP (40 +/- 9 mm Hg), greater RVH, and more severe pulmonary arterial neointimal formation than rats that received triptolide (0.25 mg/kg every other day, n = 7, mPAP 30 +/- 4 mm Hg) and rats that received triptolide (0.2 mg/kg daily, n = 7, mPAP 25 +/- 5 mm Hg, p < 0.01). In pneumonectomized rats that receive MCT, triptolide attenuates the development of pulmonary hypertension and RVH, and promotes regression of pulmonary arterial neointimal formation.

Topics: Analysis of Variance; Animals; Disease Models, Animal; Diterpenes; Drug Evaluation, Preclinical; Epoxy Compounds; Hemodynamics; Hypertension, Pulmonary; Immunosuppressive Agents; Male; Monocrotaline; Neovascularization, Pathologic; Phenanthrenes; Pneumonectomy; Rats; Rats, Sprague-Dawley; Specific Pathogen-Free Organisms; Time Factors; Tunica Intima

We compared the efficacy of azithromycin to the clinical antimalarial doxycycline in Plasmodium berghei-infected mice and in P. falciparum-infected Aotus monkeys. When mice were administered drug orally twice a day for three days, the minimum total dose of azithromycin that cured all mice was 768 mg/kg. Doxycycline at a dose of 1,536 mg/kg cured no mice. The efficacy of fast-acting blood schizonticides (quinine, halofantrine, artemisinin) against P. berghei was augmented by azithromycin. In monkey experiments in which there were two animals per experimental group, azithromycin (100 mg/kg/day for seven days) eliminated parasitemia; azithromycin (30 mg/kg/day) initially cleared 99.8-100% of the parasites with recrudescence in the one completely cleared case. Doxycycline (30 mg/kg/day) cleared 100% of the parasites with recrudescence in both cleared cases. Since azithromycin can be clinically administered at a somewhat higher daily dosage than doxycycline, the data suggest that it may be possible to replace drugs of the tetracycline class with azithromycin in combination with fast-acting blood schizonticides for the treatment of P. falciparum infection.

Topics: Administration, Oral; Animals; Antimalarials; Aotus trivirgatus; Artemisinins; Azithromycin; Disease Models, Animal; Doxycycline; Drug Therapy, Combination; Humans; Injections, Subcutaneous; Malaria; Malaria, Falciparum; Mice; Parasitemia; Phenanthrenes; Plasmodium berghei; Quinine; Sesquiterpenes

The anti-inflammatory action of nonapeptide fragments of uteroglobin or lipocortin I known as antiflammins, was tested in the carrageenan or phospholipase A2 rat paw oedema model. The development of carrageenan-induced oedema in rats was significantly inhibited during the early and late phases of the oedema by the local administration of antiflammins 1 and 2. However, the peptides were not able to inhibit phospholipase A2-induced oedema. The time course of the anti-oedematous activity of nonapeptides after intradermal carrageenan injection may be attributed to their effect on mast cell degranulation and accumulation and activation of leukocytes. Naja naja phospholipase A2 exhibited strong histamine release-inducing activity, which may have contributed to the rat paw oedema induction. Surprisingly, antiflammins had a limited but significant inhibitory effect on histamine secretion.

Topics: Animals; Annexin A1; Aristolochic Acids; Carrageenan; Cell Degranulation; Chlorpheniramine; Dexamethasone; Disease Models, Animal; Edema; Elapid Venoms; Elapidae; Histamine Release; Indomethacin; Inflammation; Injections, Intradermal; Male; Mast Cells; Peptide Fragments; Phenanthrenes; Phospholipases A; Phospholipases A2; Rats; Rats, Sprague-Dawley; Uteroglobin

This report describes, illustrates, and validates the major features of a procedure designed to provide primary assessments of the activities of potential antimalarial drugs against infections with chloroquine-resistant or pyrimethamine-resistant strains of Plasmodium falciparum in owl monkeys of Colombian origin. Studies with 14 specially selected compounds have shown that the test method has the capacity to identify and quantify diverse levels of therapeutic efficacy among agents that differ widely in chemical structure. Extended studies with two of the above compounds indicate that such assessments have an acceptable level of reproducibility. Experiments with two other agents, structurally different from those in the selected group, have shown that the impacts of pyrimethamine resistance (or chloroquine resistance) on the activity of a compound can be readily identified during routine application of the test procedure, as can emergence of parasites resistant to the test agent. The above body of information can usually be acquired in infections with two strains of P. falciparum, one chloroquine-resistant, the other pyrimethamine-resistant, with commitments of no more than 1.5 g of test compound and 12 owl monkeys. These modest requirements have made it possible to utilize human plasmodial infections in the owl monkey in the search for new blood schizonticidal drugs more broadly effective than those currently available.

Topics: Animals; Antimalarials; Aotus trivirgatus; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Drug Resistance; Female; Guanidines; Haplorhini; Hydroxamic Acids; Malaria; Male; Phenanthrenes; Plasmodium falciparum; Plasmodium vivax; Pyridines; Pyrimidines; Quinazolines; Quinolines; Sulfones; Triazines

An antimalarial drug testing system is described which utilizes trophozoite induced Plasmodium cynomolgi malaria in rhesus monkeys. The schizonticidal activity of standard antimalarial drugs in this system is reported. The system accurately predicted antimalarial activity in man of 8 of 9 compounds selected for clinical trials.

Topics: Amodiaquine; Animals; Antimalarials; Dapsone; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Drug Tolerance; Haplorhini; Macaca mulatta; Malaria; Phenanthrenes; Primaquine; Prodigiosin; Pyrimethamine; Quinine; Sulfadiazine; Sulfadimethoxine; Sulfalene; Trimethoprim

Topics: Amides; Animals; Anti-Arrhythmia Agents; Arrhythmias, Cardiac; Cats; Disease Models, Animal; Dogs; Epinephrine; Ethylamines; Female; Male; Phenanthrenes; Procainamide; Quinidine

Topics: Animals; Disease Models, Animal; Drug Synergism; Encephalomyocarditis virus; Female; Injections, Intraperitoneal; Injections, Intravenous; Leukocyte Count; Male; Mice; Muramidase; Orthomyxoviridae; Orthomyxoviridae Infections; Phagocytosis; Phenanthrenes; Poliomyelitis; Salmonella Infections, Animal; Salmonella typhimurium; Staphylococcal Infections; Staphylococcus

Topics: Animals; Antineoplastic Agents; Benz(a)Anthracenes; Castration; Disease Models, Animal; Female; Lactones; Mammary Neoplasms, Experimental; Phenanthrenes; Rats; Testolactone; Testosterone