morphinans and Inflammation

Autoimmune diseases (ADs), with significant effects on morbidity and mortality, are a broad spectrum of disorders featured by body's immune responses being directed against its own tissues, resulting in chronic inflammation and tissue damage. Sinomenine (SIN) is an alkaloid isolated from the root and stem of Sinomenium acutum which is mainly used to treat pain, inflammation and immune disorders for centuries in China. Its potential anti-inflammatory role for treating immune-related disorders in experimental animal models and in some clinical applications have been reported widely, suggesting an inspiring application prospect of SIN. In this review, the pharmacokinetics, drug delivery systems, pharmacological mechanisms of action underlying the anti-inflammatory and immunomodulatory effects of SIN, and the possibility of SIN as adjuvant to disease-modifying anti-rheumatic drugs (DMARDs) therapy were summarized and evaluated. This paper aims to reveal the potential prospects and limitations of SIN in the treatment of inflammatory and immune diseases, and to provide ideas for compensating its limitations and reducing the side effects, and thus to make SIN better translate to the clinic.

Topics: Animals; Anti-Inflammatory Agents; Immunity; Inflammation; Morphinans

The rhizome of Chinese medical plant QingTeng (scientific name: Sinomenium acutum (Thunb.) Rehd. et Wils.) is widely used by traditional medical doctors for anti-inflammation and immunoregulatory in China and other Asian countries.. The purpose of this study was to evaluate the effects and possible mechanisms of sinomenine resistance against DSS-induced inflammation in vitro and in vivo.. The UC model was induced by treating female mice with 3% DSS in vivo and human colonic epithelial cells (Hcoepic) with 0.8 mg/ml DSS in vitro. The mice and Hcoepic were then treated with sinomenine. Inflammatory factors were detected using ELISA and qRT-PCR. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 and 14-3-3θ were analyzed by bioinformatic analysis and verified by western blotting, immunofluorescent staining or immunohistochemistry.. DSS-induced Hcoepic underwent high inflammation and oxidative stress conditions, whereas sinomenine reduced the uncontrolled immune microenvironment by suppressing NF-κB signaling and targeting 14-3-3θ. Knockdown of 14-3-3θ decreased the protective effect of sinomenine against DSS-induced inflammation in vitro. Moreover, sinomenine promoted 14-3-3θ protein expression and inhibited NF-κB p65 signaling in DSS-induced mice.. These findings suggest that 14-3-3θ plays an important role in sinomenine against DSS treatment, and sinomenine could be considered a potential drug for the treatment of UC.

Topics: Animals; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Humans; Inflammation; Mice; Mice, Inbred C57BL; Morphinans; NF-kappa B; Signal Transduction

Topics: Animals; Apoptosis; Chemical and Drug Induced Liver Injury; Inflammation; Liver; Male; Mice; Mice, Inbred BALB C; Morphinans; Oxidative Stress

Mycoplasma pneumoniae (MPP) induced pneumonia is a common disease of children. Sinomenine (SIN) is an isoquinoline mainly sequestered from Sinomenium acutum. It is a promising drug for treating arthritis, lung, colon, liver and gastric cancer. Hence, the present study investigated the role and mechanism of SIN treatment in MPP induced pneumonia in experimental in-vivo mice model. The BALB/c male mice were separated into four groups (n = 6 mice/group): normal, MPP, MPP + SIN (20 mg/kg bw), and SIN (20 mg/kg bw) alone. Results were expressed as mean ± SD. Data were analyzed using one way Analysis of Variance (ANOVA) with the Dunnett's post hoc test using SPSS v 18.0. P value < 0.05 was considered significant. The total protein, cell count, inflammatory cytokines, MP-IgM, Monocyte chemo attractant protein-1 (MCP-1), and MP-DNA were measured. The protein expressions of Bax/Bcl-2, ERK, JNK, NF-κB were analyzed and histopathology of lungs was examined. SIN treatment significantly (p < 0.05) reduced the total proteins, cell counts in BALF, inflammatory cytokines, MP-IgM, MCP-1, MP-DNA and reversed the histological alterations. SIN attenuated the apoptotic pathway through the modulation of Bax/Bcl-2 expression. SIN alleviated pulmonary inflammatory mediators and apoptosis in MPP-infected mice via suppression of ERK/JNK/NF-κB signaling. SIN administration diminished inflammation and lung fibrosis by inhibiting apoptosis in MPP mice. Hence, SIN is a potential natural protective remedy for MPP.

Topics: Animals; bcl-2-Associated X Protein; Cytokines; Immunoglobulin M; Inflammation; Male; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Morphinans; Mycoplasma pneumoniae; NF-kappa B; Pneumonia, Mycoplasma

Gestational diabetes mellitus (GDM) is a dangerous complication of pregnancy which is induced via dysfunction in glucose metabolism during pregnancy. Sinomenine (SM) has already proved an antidiabetic effect against streptozotocin (STZ)-induced diabetes mellitus (DM) in rats. In this protocol, we examined the protective effect of SM against STZ-induced GDM in rats. Wistar rats were divided into three groups and STZ (40 mg/kg) was used to induce GDM. At the end of the experimental protocol, bodyweight, pub weight, and survival rate were estimated. Blood glucose level (BGL), fasting insulin (FINS), free fatty acid (FFA), Hemoglobin A1C (HbA1c), and C-peptide were measured. Lipid, antioxidant, inflammatory cytokines, and inflammatory mediators were also determined. RT-PCR was used for estimation of the role of TLR4/MyD88/NF-κB signaling pathway. SM treatment significantly (p < .001) reduced BGL, hepatic glycogen, and improved the levels of FINS, C-peptide, FFA, and HbA1c. SM significantly (p < .001) suppressed the levels of total cholesterol (TC), low-density lipoprotein (LDL), triglycerides (TG), coronary artery index (CAI), very low-density lipoprotein (VLDL), atherogenic index (AI), and boosted high-density lipoprotein (HDL) levels. SM significantly (p < .001) decreased the lipid peroxidation (LPO) level and enhanced glutathione peroxidase (GPx), total antioxidant capacity (TAC), glutathione S-transferase (GST), superoxide dismutase (SOD), respectively. It reduced the levels of inflammatory cytokines including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inflammatory mediators viz., nuclear kappa B factors (NF-κB). SM significantly (p < .001) reduced the mRNA expression of Myd88, NLRP3, TLR4, and NF-κB, which were boosted in the GDM group rats. These findings suggest that SM could be a probable drug to be used for treating GDM via inhibition of the TLR4 signaling pathway. PRACTICAL APPLICATIONS: It is well known that gestational diabetes mellitus (GDM) is a dangerous health problem during the pregnancy. SM reduced the glucose level; boosted the level of fasting insulin (FINS) and bodyweight. SM significantly improved the number of pubs and their survival rates. SM suppressed oxidative stress and inflammation via activation of TLR4/MyD88/NF-κB signaling pathway. According to our research, SM can be used as a preventive drug in the treatment of GDM during pregnancy.

Topics: Animals; Diabetes, Gestational; Female; Inflammation; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Oxidative Stress; Pregnancy; Rats; Rats, Wistar; Signal Transduction; Toll-Like Receptor 4

Sepsis is a systemic inflammatory response syndrome caused by a host's immune response to infection. Acute lung injury (ALI) is one of the most common complications of sepsis with high mortality and morbidity. Recent evidence demonstrated that the 'gut-lung axis' was related to the progression of septic acute lung injury, which regarded gut microbiota and intestinal barrier as two critical factors correlated with acute lung injury. Sinomenine is an isoquinoline alkaloid component extracted from Sinomenium acutum Rehd，et Wils, which has been already reported to have significant anti-inflammatory, immunosuppressive, and anti-arthritis properties. In this research, we observed that sinomenine could repair the lung injury and alleviate inflammatory response induced by cecum ligation and puncture (CLP). Illumine sequencing of 16S rDNA revealed that sinomenine could improve the richness of gut microbiota and modulate the composition of intestinal flora in cecum ligation and puncture mice. Meanwhile, sinomenine could reduce the colon pathological damage and improve the intestine barrier integrity in cecum ligation and puncture mice. We also found that the molecular mechanism of sinomenine's protective effect on intestinal tract was related to the activation of aryl hydrocarbon receptor/nuclear factor erythroid-2 related factor 2（Nrf2）pathway both in vivo and vitro experiments. Collectively, the prevention of septic acute lung injury by sinomenine might be mediated by modulating gut microbiota and restoring intestinal barrier via aryl hydrocarbon receptor/Nrf2-dependent pathway.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Basic Helix-Loop-Helix Transcription Factors; Cecum; Cell Line, Tumor; Cytokines; Disease Models, Animal; Gastrointestinal Microbiome; Gastrointestinal Tract; Homeostasis; Humans; Inflammation; Ligation; Male; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; Permeability; Protective Agents; Punctures; Receptors, Aryl Hydrocarbon

Atherosclerosis (AS) is a cardiovascular disease that arise due to dysfunction of lipid deposition and metabolism. AS is causes the mortality and morbidity worldwide. Sinomenine isolated from the Sinomenium acutum is used extensively against the various cardiac diseases in China. However, the anti-atherosclerosis effect of sinomenine still not explore. In this study, we explore the cardioprotective and anti-atherosclerosis effect of sinomenine against Vitamin D3 and High fat induced atherosclerosis in rats. Sprague Dawley (SD) rats were used in this study. The rats were received the vitamin D (60000) and High fat diet to induce the atherosclerosis and divided into groups and received the oral administration of sinomenine (2.5, 5 and 10 mg/kg) and simvastatin (5 mg/kg). Body weight, organ weight and biochemical parameters were estimated. The mRNA expression of MyD88, TLR4, NF-κB and IκB were estimated. Sinomenine treated rats significantly (p<0.001) suppressed the body weight and modulated the organ weight (hepatic, renal and heart). Sinomenine significantly (p<0.001) decreased the level of triacylglycerols (TG), low density lipoprotein cholesterol (LDL-c), total cholesterol (TC), very low-density lipoprotein cholesterol (VLDL-c) and augmented the level of high-density lipoprotein cholesterol (HDL-c). Sinomenine treatment also reduced the level of atherogenic index (TC/HDL-c and LDL-c/HDL-c). Sinomenine treatment decrease the ratio of HMG CoA/Mevalonate and level of collagen and total protein. Sinomenine significantly (p<0.001) altered the level of heart parameters, antioxidant parameters and inflammatory cytokines. Sinomenine significantly (p<0.001) reduced the expression of MyD88, TLR4, NF-κB and IκB. Taken together, sinomenine exhibited the protective effect against the atherosclerosis via alteration of TLR4/NF-κB signaling pathway.

Topics: Administration, Oral; Animals; Antioxidants; Atherosclerosis; Cholecalciferol; Cytokines; Diet, High-Fat; Inflammation; Inflammation Mediators; Lipid Metabolism; Male; Morphinans; NF-kappa B; Oxidative Stress; Phytotherapy; Rats, Sprague-Dawley; Signal Transduction; Sinomenium; Toll-Like Receptor 4

The objective of study was to investigate the inhibitory effect of sinomenine on neuropathic pain on dorsal root ganglia (DRG). The DRG cell line and spinal nerve ligation (SNL) model were used in this study. The effect of sinomenine on the cell viability was examined by MTT assay. The expression of p38 MAPK, NF-κB, c-fos, SP and TNF-α was detected by using immunofluorescence and immunohistochemistry assay. We also assessed the level of p-CaMKII, COX-2, p-CREB, IL-17A, TLR4 and IL-1β via western blotting and RT-qPCR. Compared to the controls, sinomenine showed a protective effect on TNF-α-induced apoptosis on DRG cells in a dose-dependent manner, with an increase of cell viability and a decrease of reactive oxygen species level as well as LDH release. Parallelly, sinomenine treatment significantly reduced the expression of various factors related to stress and inflammation, including p38 MAPK, NF-κB, c-fos, p-CAMKII, COX-2, p-CREB, TLR4 and IL-17A in DRG cells in vitro. Furthermore, we found that administration of sinomenine significantly reduced mechanical withdrawal threshold and thermal withdrawal latency and inhibited the inflammation and activation of p38 signaling in SNL rats. It is noting that combined therapy of sinomenine and pulsed radiofrequency exhibited higher efficacy of dorsal root ganglia inflammation than single treatment as well as the combination of oxycodone and pulsed radiofrequency. Sinomenine inhibited the apoptosis of DRG cell by regulating p38 MAPK/CREB signalling pathway, which provides evidence to alleviate neuropathic pain in clinic.

Topics: Analgesics; Analgesics, Opioid; Animals; Anti-Inflammatory Agents; Apoptosis; Behavior, Animal; Cell Line; Combined Modality Therapy; Cyclic AMP Response Element-Binding Protein; Disease Models, Animal; Ganglia, Spinal; Inflammation; Inflammation Mediators; Male; Morphinans; Neuralgia; Oxycodone; p38 Mitogen-Activated Protein Kinases; Pain Threshold; Pulsed Radiofrequency Treatment; Rats, Sprague-Dawley; Signal Transduction

Corneal neovascularization (CNV) causes higher-order aberrations, corneal edema, ocular inflammation, and corneal transplant rejection, thereby decreasing visual acuity. In this study, we investigated the effects of topical administration of the kappa opioid receptor agonist nalfurafine (TRK-820) on CNV. To induce CNV, intrastromal corneal sutures were placed on the corneal stroma of BALB/c mice for 2 weeks. Nalfurafine (0.1 µg/2 μL/eye) was topically administered to the cornea once or twice daily after CNV induction. The CNV score, immune cell infiltration, and mRNA levels of angiogenic and pro-inflammatory factors in neovascularized corneas were evaluated using slit-lamp microscopy, immunohistochemistry, flow cytometry, and polymerase chain reaction. The mRNA expression of the kappa opioid receptor gene Oprk1 was significantly upregulated following CNV induction. Topical administration of nalfurafine twice daily significantly suppressed CNV and lymphangiogenesis, as well as reduced the mRNA levels of angiogenic and pro-inflammatory factors in the neovascularized corneas. Moreover, nalfurafine administration twice daily reduced the numbers of infiltrating leukocytes, neutrophils, macrophages, and interferon-γ-producing CD4

Topics: Administration, Topical; Animals; Corneal Edema; Corneal Neovascularization; Corneal Stroma; Gene Expression; Inflammation; Macrophages; Male; Mice; Mice, Inbred BALB C; Morphinans; Neutrophils; Receptors, Opioid, kappa; RNA, Messenger; Spiro Compounds

Although sinomenine (SIN) has been used to treat several inflammation-related diseases in the clinic for decades, the detailed anti-inflammatory mechanism remains elusive. Here, we present a chemoproteomic study that supports a polypharmacological mode of action for SIN to inhibit inflammation. Notably, functional validation revealed multiple new protein regulators whose knockdown could significantly affect inflammation.

Topics: Animals; Anti-Inflammatory Agents; Cell Survival; Dose-Response Relationship, Drug; Inflammation; Lipopolysaccharides; Mice; Molecular Structure; Morphinans; Proteomics; RAW 264.7 Cells

Our present study focused on assessing whether Sinomenine (SIN) could attenuate sepsis-induced acute lung injury (ALI).. The mice were conditioned with SIN 1 h before intraperitoneal injection of lipopolysaccharide (LPS). Lung wet/dry (W/D) ratio, inflammatory level in bronchoalveolar lavage fluid (BALF), malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity and inflammatory cytokines production were detected. The expression of nuclear factor erythroid 2-like 2 (Nrf2) and autophagy-related proteins were detected by Western blot and immunohistochemical analyses. In addition, the RAW264.7 cells were treated with SIN 1 h before treatment with LPS. Inflammatory cytokines, iNOS and COX2 were detected. The expression of Nrf2 and autophagy-related proteins were explored by Western blot analysis.. Experiments in vivo and in vitro discovered that LPS significantly increased the degree of injury, inflammatory cytokines production and oxidative stress. However, the increase was significantly inhibited by treatment of SIN. In addition, SIN was found to upregulate the expression of Nrf2 and autophagy-related proteins both in vivo and in vitro.. Our data suggested that SIN could attenuate septic-associated ALI effectively, probably due to the inhibition of inflammation and oxidative stress through Nrf2 and autophagy pathways.

Topics: Acute Lung Injury; Animals; Autophagy; Cytokines; Female; Inflammation; Kelch-Like ECH-Associated Protein 1; Lipopolysaccharides; Mice; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; Oxidative Stress; RAW 264.7 Cells; Sepsis

Morphine and its derivatives play inevitably important role in the μ-opioid receptor (MOR) targeted antinociception. A structure-activity relationship study is presented for novel and known orvinol and thevinol derivatives with varying 3-O, 6-O, 17-N and 20-alkyl substitutions starting from agonists, antagonists and partial agonists. In vitro competition binding experiments with [

Topics: Analgesics, Opioid; Animals; Behavior, Animal; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Guinea Pigs; Inflammation; Male; Molecular Docking Simulation; Molecular Structure; Morphinans; Osteoarthritis; Rats, Wistar; Receptors, Opioid, mu; Structure-Activity Relationship

Amyloid beta is a major constituent of the plaques found in the brains of patients suffering from Alzheimer's disease (AD). A growing body of research work suggests that neuroinflammation plays important roles in the development of AD. Thus, considerable efforts are directed towards identification of compounds that can reduce or inhibit neuroinflammation. Here, we show that sinomenine, a compound present in a Chinese medicinal plant, Sinomenium acutum, inhibits oligomeric amyloid beta-induced production of reactive oxygen species (ROS), nitric oxide (NO) and inflammation-related molecules from astrocytic cells. The conditioned medium from oligomeric amyloid beta-treated astrocytic cells induces cell death in the hippocampal neuronal cells. Importantly, sinomenine inhibits this cell death. In addition, this compound has inhibitory effects on the production of ROS, NO and inflammation-related factors from oligomeric amyloid-beta treated human astrocytes. Finally, the conditioned medium from oligomeric amyloid beta-treated human astrocytes induces cell death in the primary culture of human neurons, which is inhibited by sinomenine. Thus, sinomenine inhibits amyloid beta-induced production of toxic factors from astrocytes, and confers protection to hippocampal neuronal cells as well as human neurons against indirect toxicity. The results suggest that this compound could provide beneficial effects in AD and other neurodegenerative conditions by reducing inflammation and neuronal cell death.

Topics: Amyloid beta-Peptides; Animals; Astrocytes; Cell Line; Hippocampus; Humans; Inflammation; Mice; Morphinans; Neurons; Neuroprotective Agents; Nitric Oxide; Reactive Oxygen Species

Oxidative stress, inflammation and pyroptosis are three of the most important mechanisms in the pathophysiology of temporal lobe epilepsy (TLE). Most people with TLE are refractory to the existing drugs. Sinomenine has shown neuroprotective effects through counteracting oxidative stress, inflammation and pyroptosis. In this study, we evaluated the effect of sinomenine on seizure behavior, oxidative stress, inflammation and pyroptosis markers in addition to its neuroprotective potential in intrahippocampal kainate-induced rat model of TLE. For this purpose, male rats (n = 60) were randomly divided into five groups, i.e., sham, kainate (lesion) with an intrahippocampal injection of kainate, kainate groups receiving sinomenine at doses of 30 or 50 mg/kg, and kainate group receiving valproic acid at a dose of 200 mg/kg (as the positive control). Our obtained data showed that sinomenine administration at a dose of 50 mg/kg can significantly decreases severity of seizures and incidence of status epilepticus (SE), hippocampal aberrant MFS and DNA fragmentation and prevents reduction of neuronal density. It also significantly restored level of ROS, MDA, HO-1 and SOD but its effect on GSH level was not significant. Additionally, sinomenine at a dose of 50 mg/kg partially counteracted the increase of NF-κB, TLR 4, TNFα, GFAP and caspase 1. These results suggest that sinomenine has anticonvulsant and neuroprotective effects by reducing hippocampal oxidative stress, inflammation, pyroptosis and apoptosis in intrahippocampal kainate model of TLE.

Topics: Animals; Anticonvulsants; Disease Models, Animal; Epilepsy, Temporal Lobe; Hippocampus; Inflammation; Kainic Acid; Male; Morphinans; Neuroprotective Agents; Oxidative Stress; Pyroptosis; Rats; Reactive Oxygen Species

Liver disease is common and often life-threatening. Sinomenine (SIN) is an active ingredient extracted from . In vivo experiments, mice were randomly divided into six groups (n=10): control group, model group, SIN (25 mg/kg) group, SIN (50 mg/kg) group, SIN (100 mg/kg) group and SIN (100 mg/kg) + SRI-011381 group. Alanine transaminases (ALT), aspartate transaminases (AST) and alkaline phosphatase (ALP) were detected. The pathological lesion was measured by HE staining. Apoptosis was measured by TUNEL staining. In vitro experiments, BRL-3A cells were treated with APAP (7.5 mM) and then subjected to various doses of SIN (10, 50 and 100 μg/mL) at 37°C for 24 h. Inflammatory factors and oxidative stress index were measured by ELISA. The expression of proteins was detected by Western blot.. The results showed that compared with the control group, the levels of ALT, AST and ALP in the serum of APAP-induced mice were significantly increased, followed by liver histological damage and hepatocyte apoptosis. Besides, APAP reduced the activity of SOD and GSH-Px, while increasing the content of MDA and LDH. Notably, APAP also promoted the expression of NLRP3, ASC, caspase-1 and IL-1β. Interestingly, SIN treatment dose-dependently reduced APAP-induced liver injury and oxidative stress, inhibited the activation of NLRP3 inflammasomes, and reduced the levels of inflammatory cytokines. In vitro studies have shown that SIN treatment significantly reduced the viability of BRL-3A cells and oxidative stress and inflammation. In addition, the Western blotting analysis showed that SIN inhibited the activation of TGF-β/Smad pathway in a dose-dependent manner in vitro and in vivo. These effects were significantly reversed by TGF-β/Smad activator SRI-011381 or TGF-β overexpression.. The study indicates that SIN attenuates APAP-induced acute liver injury by decreasing oxidative stress and inflammatory response via TGF-β/Smad pathway in vitro and in vivo.

Topics: Acetaminophen; Animals; Apoptosis; Cells, Cultured; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Inflammation; Mice; Mice, Inbred C57BL; Morphinans; Oxidative Stress; Smad Proteins; Structure-Activity Relationship; Transforming Growth Factor beta

Sinomenine (SIN) is clinically used as an anti-rheumatic drug. However, the metabolic and pharmacological mechanisms of SIN combined with its metabolites are unclear. This study aims to explore the cyclic metabolic mechanism of SIN, the anti-inflammation effects of SIN and its major metabolites (N-demethylsinomenine (DS) and sinomenine-N-oxide (SNO)), and the oxidation property of SNO.. SIN was administrated to rats via gavage. Qishe pills (a SIN-containing drug) were orally administrated to humans. The bio-samples were collected to identify SIN's metabolites. Enzymatic and non-enzymatic incubations were used to reveal SIN's metabolic mechanism. Impacts of SIN, SNO and DS on the inflammation-related cytokine's levels and nuclear translocation of NF-κB were evaluated in LPS-induced Raw264.7 cells. ROS induced by SNO (10 μM) was also assessed.. CYP3A4 and ROS predominantly mediated the formation of SNO, and CYP3A4 and CYP2C19 primarily mediated the formation of DS. Noteworthily, SNO underwent N-oxide reduction both enzymatically, by xanthine oxidase (XOD), and non-enzymatically, by ferrous ion and heme moiety. The levels of IL-6 and TNF-α and nuclear translocation of NF-κB were ameliorated after pretreatment of SIN in LPS-induced Raw264.7 cells, while limited attenuations were observed after pretreatment of DS (SNO) even at 200 μM. In contrast, SNO induced ROS production.. This study elucidated that SIN underwent both enzymatic and non-enzymatic cyclic metabolism and worked as the predominant anti-inflammation compound, while SNO induced ROS production, suggesting more studies of SIN combined with SNO and DS are necessary in case of DDI and potential toxicities.

Topics: Animals; Anti-Inflammatory Agents; Female; Humans; Inflammation; Male; Mice; Microsomes, Liver; Morphinans; Oxidative Stress; Rats; Rats, Sprague-Dawley; RAW 264.7 Cells

Traumatic spinal cord injury (SCI) is a devastating condition with few efficacious drugs. Sinomenine, a bioactive alkaloid extracted from medicinal herb, has been used as a treatment of rheumatoid diseases. This present study explored the therapeutic effects of sinomenine on locomotor dysfunction and neuropathology in SCI. Our findings revealed that sinomenine mitigated neurological deficits and enhanced neuronal preservation, paralleled with a reduction of apoptosis. Also, sinomenine significantly reduced inflammatory cytokines and oxidative stress factors. We further examined erythroid-2-related factor 2 (Nrf2) nuclear translocation, which mainly controls the coordinated expression of important antioxidant and detoxification genes. An increase in Nrf2 translocation from cytoplasm to nucleus and Nrf2-mediated transactivation was observed after sinomenine administration. Knocking down Nrf2 by siRNA could counteract sinomenine-mediated anti-oxidant stress and anti-inflammation following H

Topics: Animals; Female; Hydrogen Peroxide; Inflammation; Morphinans; NF-E2-Related Factor 2; Oxidative Stress; PC12 Cells; Rats; Rats, Sprague-Dawley; Signal Transduction; Spinal Cord Injuries; Thoracic Vertebrae

The initiation of pressure ulcers is accompanied by inflammation. Sinomenine emerges as a potential anti-inflammation agent. The aim of this study was to corroborate its anti-inflammatory property in skin keratinocyte HaCaT cells. Long non-coding RNA colon cancer associated transcript-1 (CCAT1)-associated mechanisms were also investigated.. HaCaT cells were stimulated with lipopolysaccharide (LPS) for 6 h after sinomenine pre-administration. Transfection was carried out to induce CCAT1 overexpression or silence it in HaCaT cells. Viability and apoptosis of HaCaT cells were determined by MMT and observed using flow cytometry, respectively. Protein expression was quantified using Western blot or ELISA. CCAT1 was measured by qRT-PCR.. LPS notably decreased cell viability and exaggerated apoptosis with the cleavage of caspase-3/-9. The secretion of inflammatory factors was promoted. Sinomenine pre-administration maintained cell viability, blocked apoptosis and relieved inflammation with the decrease in cleaved caspase-3/-9 and inflammatory factors. LPS-induced phosphorylation of p65, IκBα and p38MAPK and overexpression of CCAT1 were precluded by sinomenine. CCAT1 overexpression, which per se induced inflammatory lesions, negated the positive effects of sinomenine with the restored phosphorylation of p65, IκBα, and p38MAPK.. Sinomenine played a protective role against LPS-induced inflammation. The anti-inflammatory activity of sinomenine might be mediated by CCAT1 down-regulation.

Topics: Anti-Inflammatory Agents; Antirheumatic Agents; Cells, Cultured; Down-Regulation; Gene Expression Regulation; Humans; Inflammation; Keratinocytes; Lipopolysaccharides; Morphinans; RNA, Long Noncoding; Skin

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal).\ \ This article has been retracted at the request of the Editor-in-Chief.\ \ Given the comments of Dr Elisabeth Bik regarding this article “This paper belongs to a set of over 400 papers (as per February 2020) that share very similar Western blots with tadpole-like shaped bands, the same background pattern, and striking similarities in title structures, paper layout, bar graph design, and - in a subset - flow cytometry panels”, the journal requested the authors to provide the raw data. However, the authors were not able to fulfil this request and therefore the Editor-in-Chief decided to retract the article.

Topics: Animals; Apoptosis; Apoptosis Regulatory Proteins; Cell Survival; Down-Regulation; Dual Specificity Phosphatase 1; Female; Guinea Pigs; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; Lipopolysaccharides; Male; MAP Kinase Signaling System; MicroRNAs; Morphinans; Signal Transduction; Tumor Necrosis Factor-alpha; Up-Regulation

BACKGROUND In the present study, we aimed to investigate the effects of sinomenine (SIN) on chronic intermittent hypoxia (CIH)- induced lung injury in rats, and to explore the underlying mechanisms. MATERIAL AND METHODS To perform the investigation, a CIH rat model was established. ELISA assay was applied to detect the level of inflammatory cytokines. Oxidative stress bio-markers (MDA, SOD, and CAT) were determined in lung tissues. In addition, the expression level of NADPH oxidase 2 (Nox2) was analyzed by Western blotting and qRT-PCR, respectively. RESULTS The results showed that compared with other groups, more obvious pulmonary pathological changes were observed in the CIH group. The level of inflammatory cytokines in the CIH group was markedly higher than that in the control and Con-S groups. Compared with the control and Con-S groups, oxidative stress was notably increased in the CIH group. Expression of Nox2 was also increased in the CIH group. The effects caused by CIH in rats were attenuated by SIN treatment. CONCLUSIONS SIN can reverse chronic intermittent hypoxia-induced lung injury through inhibiting inflammation and oxidative stress.

Topics: Animals; Chronic Disease; Cytokines; Down-Regulation; Hypoxia; Inflammation; Inflammation Mediators; Lung; Lung Injury; Male; Mice; Morphinans; NADPH Oxidases; NF-E2-Related Factor 2; Oxidative Stress; Rats, Sprague-Dawley; Signal Transduction

BACKGROUND The purpose of this study was to investigate the effects of sinomenine (SIN) on CFA-induced inflammatory pain in rats, and to explore the underlying molecular mechanisms. MATERIAL AND METHODS To determine the potential influences of SIN in the pathogenesis of inflammatory pain, an inflammatory pain (IP) mouse model was established and rats were treated with SIN (30 mg/kg). Behavioral tests were used to assess the MWT and TWL of the rats. ELISA assay was used to detect the level of inflammation cytokines. Western blotting and qRT-PCR were carried out to measure the related protein and mRNA expression level, respectively. RESULTS We found that the MWT and TWL of the CFA-treated rats were markedly lower than that of the control rats, and they were significantly increased by SIN administration. The results suggest that IP rats had higher levels of TNF-α, IL-1β and IL-6 compared with the control rats. SIN administration decreased the levels of TNF-α, IL-1β, and IL-6. In addition, we found that p-p65 and p-p38 expression notably decreased after SIN treatment in IP rats. Moreover, the results showed that SIN inhibited Cox-2 and PGE2 expression in IP rats. CONCLUSIONS The data indicate that SIN had a protective role in inflammatory pain through repressing inflammatory mediators via preventing the p38MAPK-NF-κB pathway.

Topics: Animals; Cyclooxygenase 2; Cytokines; Freund's Adjuvant; Inflammation; Interleukin-1beta; Male; Morphinans; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Pain; Rats; Rats, Sprague-Dawley; Signal Transduction; Tumor Necrosis Factor-alpha

BACKGROUND This study aimed to investigate the effect and underlying molecular mechanism of sinomenine (SIN) on ankylosing spondylitis (AS). MATERIAL AND METHODS To study the potential role of SIN in the pathogenesis of AS, an AS mouse model was established and mice were treated with different concentrations of SIN (10, 30, and 50 mg/kg, administered intraperitoneally). Markers of inflammation and oxidative stress were determined by ELISA assay. Western blot analysis and qRT-PCR were used to quantify the levels of related proteins and gene mRNA expression. RESULTS The results suggest that AS mice has higher levels of TNF-α, IL-1β, and IL-6 (p<0.01 for all), and lower levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) (p<0.01 for all). SIN treatment reduced the level of TNF-α, IL-1β, and IL-6 in a dose-dependent manner, and the levels of SOD, CAT, and GSH-PX were dose-dependently increased (p<0.05 for all). The results also revealed that NF-κBp65 expression decreased, while the level of IkB increased, in a dose-dependent manner, after SIN treatment in AS mice (p<0.05 for all). The level of p-p38 was dose-dependently reduced in AS mice by SIN treatment (p<0.05). Moreover, SIN inhibited Cox-2 expression in AS mice in a dose-dependent manner (p<0.05). CONCLUSIONS SIN has a beneficial role in AS through suppressing inflammatory mediators and by down-regulating oxidative stress via inhibiting the MAPKp38/NF-kB pathway and Cox-2 expression.

Topics: Animals; Antioxidants; Catalase; Disease Models, Animal; Glutathione Peroxidase; Inflammation; Interleukin-1beta; Interleukin-6; Mice; Mice, Inbred BALB C; Morphinans; NF-kappa B; Oxidative Stress; Signal Transduction; Spondylitis, Ankylosing; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Studies have shown that the inflammatory activation of miroglia (MG) and nuclear factor kappa B ( NF-κB ) play a dominant role in inflammatory response. Previous studies have shown that sinomenine, an anti-inflammatory agent extracted from Sinomenium acutum, can directly protect neurons against cerebral ischemia injury. However, there are no reports on its effect on ischemia/reperfusion-induced inflammatory activation of MG. In the present study, an in vitro ischemia/reperfusion model was developed with mouse BV-2 microglia cells, a model of oxygen-glucose deprivation/reperfusion (OGD/R), and the inhibitory effect of sinomenine pretreatment on inflammatory activation was confirmed through measurement of inflammatory indicators. Mechanistically, sinomenine suppressed OGD/R-induced inflammatory activation through the SP1/miRNA-183-5p/IκB-α pathway. In conclusion, this study shows that sinomenine effectively inhibits OGD/R-induced inflammatory activation in MG by suppressing the activation of transcription specificity protein 1 (SP 1). This finding is of significance for the clinical use of sinomenine in treating cerebral ischemia/reperfusion injury.

Topics: Animals; Anti-Inflammatory Agents; Cell Line; Glucose; Inflammation; Mice; Microglia; MicroRNAs; Morphinans; NF-KappaB Inhibitor alpha; Oxygen; Reperfusion Injury; Signal Transduction; Sinomenium; Sp1 Transcription Factor

Acute lung injury (ALI) is a common disease characterized by pulmonary inflammation and oxidative stress. Sinomenine (SIN) is an alkaloid originally extracted from the Chinese medicinal plant Sinomenium acutum. It has been shown to have anti-inflammatory and anti-oxidative effect. However, it's unclear whether SIN can alleviate ALI. In this study, we assessed the effect of SIN on Escherichia coli (E.coli)-induced ALI mouse model. Mice were conditioned with SIN or placebo 1 h before intratracheally instilled with E.coli. Lung water content, malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, Myeloperoxidase (MPO) levels and inflammatory cytokines production were measured. Immunohistochemistry and western blot were performed to measure target protein expression. E.coli induced histological changes indicating tissues damage and increased W/D ratio, MPO activity, MDA content, and inflammatory cytokines production in the Lung. Whereas in mice pretreated with SIN, these changes were absent. E.coli-induced NF-κB activation was also inhibited by SIN. In addition, SIN increased the expression of HO-1, NQO1 and Nrf2 in lung tissues. Our results suggest that SIN attenuates ALI through the inhibition of inflammation and oxidative stress.

Topics: Acute Lung Injury; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Escherichia coli; Female; Inflammation; Lung; Malondialdehyde; Mice, Inbred ICR; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Protective Agents; Protein Kinase C; Signal Transduction; Superoxide Dismutase

BACKGROUND This study aimed to investigate the effects of SIN on ankle fracture and the underlying mechanisms in MG-63 cells. MATERIAL AND METHODS qRT-PCR and ELISA assay were used to detect the mRNA and protein levels of cytokines in peripheral blood of children with or without ankle fracture. The expression and activity of antioxidant and detoxifying enzymes were detected by ELISA assay. Pretreated MG-63 cells with/without SIN were stimulated with 1 μg/ml bradykinin (BK). A CCK-8 kit was used to detect the cell viability. The cytokines produced from MG-63 cells were detected by Western blotting and qRT-PCR. Moreover, Western blotting was used to detect the levels of p-p38 and p-NF-κB (p65), and the activation level of the Nrf2 signaling pathway was examined by qRT-PCR and Western blotting. RESULTS In this study, we found that compared with the healthy children, the mRNA and protein levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) were significantly upregulated in children with ankle fracture. In addition, the expression and activity of antioxidant and detoxifying enzymes were imbalanced in children with ankle fracture. SIN treatment did not have a cytotoxic effect on MG-63 cells. SIN dose-dependently suppressed BK-induced upregulation of IL-1β, IL-6, TNF-α, p-p38, and p-NF-κB (p65). Furthermore, SIN dramatically inhibited oxidative stress induced by BK via balancing the expression and activity of antioxidant and detoxifying enzymes and inhibited the activation of Nrf2 signaling. CONCLUSIONS SIN might be a potential agent for the treatment of ankle fracture through reducing inflammatory response and oxidative stress.

Topics: Ankle Fractures; Ankle Joint; Cell Line, Tumor; Child; Child, Preschool; Cytokines; Female; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Male; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Phosphorylation; Reactive Oxygen Species; Signal Transduction; Tumor Necrosis Factor-alpha

To investigate the effect of sinomenine on lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages and the underlying mechanisms.
 Methods: The mouse RAW264.7 macrophages were treated with sinomenine and/or LPS with or without heme oxygenase-1 (HO-1) inhibitor Znpp. Real-time PCR, ELISA, immunofluenscence, and Western blot were used to detect the mRNA expression of TNF-α and IL-6, the release of TNF-α and IL-6, the protein expression of HO-1 and autophagy, respectively.
 Results: Compared with the control group, the mRNA expression and release of inflammatory cytokines TNF-α and IL-6 were increased, the green fluorescence of autophagy-related protein LC3 was accumulated and the protein expression of HO-1 was increased in RAW264.7 cells after LPS treatment (P<0.05). Compared with the LPS group, sinomenine treatment could reduce the mRNA expression and release of TNF-α and IL-6, accompanied by increasess in green fluorescence aggregation of LC3 and HO-1 production (P<0.05). HO-1 inhibitor Znpp could weaken the ability of sinomenine through suppressing TNF-α and IL-6 expression and decreasing the aggregation of LC3 green fluorescence (P<0.05).
 Conclusion: Sinomenine could alleviate LPS-induced inflammation in RAW264.7 macrophages, which might be related to HO-1 mediated autophagy. This study provides an experimental and theoretical basis for the clinical application of sinomenine in prevention and treatment of inflammation.. 目的：探讨青藤碱对脂多糖诱导的RAW264.7巨噬细胞炎症的影响和机制。方法：以小鼠RAW264.7巨噬细胞为研究对象，在有或无血红素氧合酶-1(heme oxygenase-1，HO-1)抑制剂Znpp处理下，采用青藤碱和/或脂多糖(lipopolysaccharide，LPS)处理RAW264.7巨噬细胞。应用Real-time PCR检测细胞炎症因子TNF-α和IL-6 mRNA表达，ELISA检测细胞炎症因子TNF-α和IL-6水平，免疫荧光试验分析细胞自噬情况，Western印迹检测细胞HO-1蛋白表达。结果：与对照组相比，LPS作用后RAW264.7细胞炎症因子TNF-α和IL-6表达和释放增多，自噬相关蛋白LC3绿色荧光聚集，HO-1表达水平升高(P<0.05)；与LPS组相比，加用青藤碱处理能减少TNF-α和IL-6表达和释放，进一步促进LC3绿色荧光聚集，增加HO-1水平(P<0.05)；用HO-1抑制剂Znpp预处理后，青藤碱对LPS诱导TNF-α和IL-6表达和释放的抑制作用减弱，LC3绿色荧光聚集减少(P<0.05)。结论：青藤碱能减轻LPS诱导的RAW264.7巨噬细胞炎症，其机制可能与HO-1介导的自噬激活有关，这为青藤碱应用于炎症反应的防治提供了实验基础和理论依据。.

Topics: Animals; Anti-Inflammatory Agents; Autophagy; Gene Expression Regulation, Enzymologic; Heme Oxygenase-1; Inflammation; Lipopolysaccharides; Macrophages; Mice; Morphinans

BACKGROUND Sinomenine (SIN) is an extract of the Chinese medicinal herb Sinomenium acutum; it has various pharmacological properties, including immunosuppression and anti-inflammation. The present study aimed to investigate whether SIN has an anti-depressant-like effect in a mouse model of depression induced by chronic unpredictable mild stress (CUMS), and to explore the underlying molecular mechanisms. MATERIAL AND METHODS A mouse model of depression was established and treated with different concentrations of SIN (30, 100, or 300 mg/kg). Then, behavioral tests, including sucrose preference test (SPT), forced swimming test (FST), and the tail suspension test (TST), were performed. The levels of norepinephrine (NE), 5-hydroxytryptamine (5-HT), and proinflammatory cytokines (interleukin-1β [IL-1β] interleukin-6 [IL-6], and tumor necrosis factor-α [TNF-α]) in the hippocampus of mice were detected by ELISA assay. The levels of p-p38, p-p65, NLRP3, ASC, and caspase-1 were measured by Western blot or/and qRT-PCR. RESULTS The results showed that SIN significantly relieved CUMSinduced depressive-like behaviors. Compared with the model mice, SIN treatment significantly increased the sucrose preference of the mice, and the immobility time in the forced swimming and the tail suspension test were shortened. In addition, SIN decreased CUMS-induced reduction in the concentrations of NE and 5-HT in the hippocampus of mice. SIN reduced CUMS-induced increases in the levels of IL-1β, IL-6, and TNF-α in the hippocampus of mice. Furthermore, activation of the p38MAPK-NF-κB pathway and the nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome induced by CUMS were inhibited by SIN treatment. CONCLUSIONS In conclusion, our results indicate the antidepressantlike effects of SIN on chronic unpredictable mild stress-induced depression in a mouse model.

Topics: Animals; Antidepressive Agents; Behavior, Animal; China; Depression; Depressive Disorder; Disease Models, Animal; Hippocampus; Inflammation; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred ICR; Morphinans; Norepinephrine; Serotonin; Stress, Psychological; Tumor Necrosis Factor-alpha

Sinomenium acutum has been used in traditional medicine to treat a painful disease such as rheumatic arthritis and neuralgia. Sinomenine, which is a main bioactive ingredient in Sinomenium acutum, has been reported to have an analgesic effect in diverse pain animal models. However little is known about the detailed mechanisms underlying peripheral analgesic effect of sinomenine. In the present study, we aimed to elucidate its cellular mechanism by using formalin-induced acute inflammatory pain model in mice. We found that intraperitoneal (i.p.) administration of sinomenine (50mg/kg) suppressed formalin-induced paw licking behavior in both the first and the second phase. Formalin-induced c-Fos protein expression was also suppressed by sinomenine (50mg/kg i.p.) in the superficial dorsal horn of spinal cord. Whole-cell patch-clamp recordings from small-sized dorsal root ganglion (DRG) neurons revealed that sinomenine reversibly increased the spike threshold and the threshold current intensity for evoking a single spike and decreased firing frequency of action potentials evoked in response to a long current pulse. Voltage-gated sodium currents (I

Topics: Animals; Antirheumatic Agents; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Formaldehyde; Ganglia, Spinal; Inflammation; Male; Mice; Mice, Inbred C57BL; Morphinans; Neuralgia; Pain; Pain Measurement; Patch-Clamp Techniques; Proto-Oncogene Proteins c-fos; Sensory Receptor Cells; Sodium; Voltage-Gated Sodium Channels

The aim of this study was to investigate in piglets, the anti-endotoxin and anti-inflammatory effects of sinomenine, an agent commonly found in Chinese herbal medicines. In high-, middle- and low-dose sinomenine groups, piglets were initially challenged with endotoxin (i.e., 1 mg lipopolysaccharide (LPS)/kg) by intraperitoneal (IP) injection and, 3 h later, intramuscularly (IM) with sinomenine at 1, 5, or 10 mg/kg. In a drug control group, piglets were dosed IP with vehicle and 3 h late IM with 10 mg/kg sinomenine while those in an LPS control group were challenged with 1 mg LPS/kg (IP) and then vehicle 3 h later; naïve control piglets were administered normal saline IP and then IM only. At 12, 24, and 48 h post-LPS/vehicle injection, blood samples were collected from the precaval vein of piglets. Clinical signs were recorded during the trial and index levels were analyzed by ELISA kits. The results revealed sinomenine could reduce the incidence/severity of certain LPS-induced toxicities, e.g., cell adhesion, systemic inflammation, and multiple organ dysfunction. Taken together, the data suggested to us that sinomenine might effectively be useful to regulate inflammatory responses as part of future anti-endotoxin therapies.

Topics: Animals; Animals, Newborn; Anti-Inflammatory Agents; Endotoxins; Inflammation; Injections, Intramuscular; Lipopolysaccharides; Morphinans; Severity of Illness Index; Swine; Treatment Outcome

Rheumatoid arthritis (RA) is a systemic autoimmune disorder mainly characterized by inflammation of the synovial tissue that can lead to destruction of bone and cartilage. Sinomenine is an alkaloid extracted from the stem of the Chinese medicinal plant Sinomenium acutum. It has been reported that sinomenine has immunosuppressive and anti-inflammatory properties. However, the molecular mechanism underlying the effect of sinominine on IL-1β-induced human RA fibroblast-like synoviocytes (RAFLS) is poorly understood. Therefore, in this study, we investigated the effect of sinomenine on the expression of inflammatory cytokines in IL-1β-treated human RAFLS in vitro and the underlying mechanism. RAFLS viability was evaluated using the MTS assay after sinomenine treatment. The levels of inflammatory cytokines were measured with ELISA, RT-PCR and western blot, respectively. The levels of TLR4 and its downstream signaling targets were determined by western blot analysis. We found that sinomenine suppressed not only NO and PGE2 production but also iNOS and COX-2 expression in IL-1β-induced RAFLS. It also inhibited the expression of TNF-α and IL-6 in IL-1β-stimulated RAFLS. Furthermore, sinomenine prevented IL-1β-induced TLR4, MyD88 and p-NF-κB p65 expression. Taken together, these results demonstrated that sinomenine prevented IL-1β-induced inflammation in human RAFLS at least in part by inhibiting the TLR4/MyD88/NF-κB signaling pathway, suggesting that sinomenine could be a potential agent in the treatment of RA.

Topics: Antirheumatic Agents; Arthritis, Rheumatoid; Blotting, Western; Cell Survival; Cells, Cultured; Cytokines; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gene Expression Regulation; Humans; Inflammation; Morphinans; Myeloid Differentiation Factor 88; NF-kappa B; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Sinomenium; Synoviocytes; Toll-Like Receptor 4

Topics: Analgesics; Animals; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Chronic Pain; Freund's Adjuvant; Gyrus Cinguli; Inflammation; Mice; Morphinans; Phosphorylation; Receptors, N-Methyl-D-Aspartate; Signal Transduction; Up-Regulation

Mast cells are vital mediators of drug allergy and, therefore, studying the relationship between drug allergy and mast cells is essential. Sinomenine is the principal active component of Sinomenium acutum, which has anti-inflammatory and anti-immune effects, and is used to treat various rheumatoid diseases. However, allergic responses to sinomenine are frequently reported. Therefore, this study assessed the effects of sinomenine on mast cell activation to characterize its allergic effects and the underlying mechanisms. Enzyme-linked immunosorbent assay (ELISA), western blot analyses, and degranulation assays were performed to measure pro-inflammatory and allergic mediators in P815 cells. The allergenic effects of sinomenine were also determined in mice by using active general anaphylaxis (ASA). The results indicated that sinomenine induced inositol-1,4,5-trisphosphate (IP

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Calcium; Cell Degranulation; Cell Line, Tumor; Drug Hypersensitivity; Inflammation; Inositol 1,4,5-Trisphosphate Receptors; Interleukin-6; Male; Mast Cells; Mice; Mice, Inbred BALB C; Morphinans; Phospholipase C gamma; Phosphorylation; Signal Transduction; src-Family Kinases; Up-Regulation

Sinomenine is originally derived from medicinal herb and used preferentially in treatment of rheumatoid diseases in Far East regions. SIN has strong anti-inflammatory and immune-regulatory properties, acting mainly through inhibiting NF-kB signaling. Although the upstream target through which SIN affects NF-kB activity is unknown, evidence suggests that SIN might regulate inflammation through Nrf2 signaling. In this study we explored the role of Nrf2 in mediating SIN's anti-inflammation and kidney protection in a mouse model of obstructive nephropathy. We found that SIN is an activator of Nrf2 signaling. It markedly increased Nrf2 protein level, Nrf2 nuclear translocation, Nef2 transcription capacity, and the downstream protein expression. We further demonstrated that SIN activation of Nrf2 is likely due to its repression of the Nrf2 inhibitor Keap1 since it drastically reduced Keap1 protein through the PKC-sensitive ubiquitination-proteasomal degradation. SIN treatment of nephropathy mice effectively reduced the kidney damage and inflammatory responses, balanced renal oxidative stress, and improved the pathological protein expression in an Nrf2 dependent manner. In addition, SIN also Nrf2-dependently modulated macrophage M1/M2 polarization and inhibited the IkBα phosphorylation and NF-kB nuclear translocation, hence revealing an important upstream event that contributed to its anti-inflammation and tissue protection. Taken together our study has identified a novel pathway through which SIN exerts its anti-inflammation and renal protective functions, and provided a molecular basis for SIN potential applications in the treatment of kidney and other inflammatory disorders.

Topics: Acute Kidney Injury; Animals; Disease Models, Animal; Heme Oxygenase-1; Humans; Inflammation; Kidney; Macrophages; Mice; Morphinans; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Signal Transduction

Intracerebral hemorrhage (ICH) causes morbidity and mortality and commonly follows the reperfusion after an ischemic event. Microglial activation mediated cytokine and protease secretion contributes to brain injury in ICH. Previous studies have shown that sinomenine possesses potent immunoregulatory properties. However, little is known about its exact role in ICH. In the present study, to investigate the effect of sinomenine on microglial cells inflammation, we treated ICH-challenged BV2 microglial cells with sinomenine in vitro, and explored its neuroprotection role in intracerebral hemorrhage in vivo. Changes in inflammatory cytokines, such as TNF-α, IL-1β and IL-6, reactive oxygen species (ROS) and NF-κB activation NF-κB were observed. In addition, the neurological deficit and cerebral water content of ICH mice were studied. The results demonstrated that sinomenine could inhibit the release of these cytokines and attenuate ROS production in a dose-dependent manner, and reduce NF-κB activation. Furthermore, sinomenine markedly inhibited cerebral water content and neurological deficit. In conclusion, our findings suggest that sinomenine played the protective effects through inhibition of microglial inflammation, and the findings also provided a novel therapy to treat ICH induced brain injury.

Topics: Animals; Brain Injuries; Cell Line; Cell Movement; Cerebral Hemorrhage; Inflammation; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Microglia; Morphinans; Neuroprotective Agents; NF-kappa B; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Sinomenine (SIN) is the active principle of the Chinese medical plant Sinomenium acutum which is widely used for the treatment of rheumatoid arthritis (RA) in China. Recently, several groups indicated that myeloid differentiation primary response protein 88 (MyD88) might be associated with disease progression of RA. Here, we observed the effect of SIN on MyD88 expression and showed its therapeutic role in RA. First, immunohistochemical staining in clinical specimens showed that MyD88 was mainly located in characteristic pathological structures of RA synovial tissues. Second, we found that MyD88 was overexpressed in the synovial tissues of the rats with adjuvant-induced arthritis (AIA). Treatment with SIN markedly decreased the expression of MyD88 in AIA rats. Finally, we provided evidences that SIN suppressed inflammation response and inflammation-induced joint destructive progression and arthritis symptoms in AIA rats. Therefore, SIN is an effective therapeutic agent for RA. Targeting MyD88 signaling may provide new methods for the treatment of RA.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Disease Progression; Drugs, Chinese Herbal; Inflammation; Joint Diseases; Male; Medicine, Chinese Traditional; Morphinans; Myeloid Differentiation Factor 88; Rats; Rats, Sprague-Dawley; Synovial Membrane; Toll-Like Receptor 2; Toll-Like Receptor 4

Sinomenine is an alkaloid originally isolated from the root of the plant Sinomenium acutum. It is used in traditional medicine in China to treat rheumatic arthritis. In the present study, we evaluated the potential antinociceptive effects of sinomenine in rodents with nociceptive, inflammatory and neuropathic pain. In normal rats and mice, systemic sinomenine produced moderate antinociceptive effect in the hot plate and tail flick tests. Sinomenine also exerted analgesic effects on mechanical and heat hypersensitivity in mice after carrageenan induced inflammation. Finally, sinomenine effectively alleviated mechanical and cold allodynia in rats and mice after injury to peripheral nerve or spinal cord. The analgesic effect of sinomenine is not associated with side effects and is not reversed by the opioid receptor antagonist naloxone. Our results showed that sinomenine has a wide spectrum analgesic effect in rodent models of nociceptive, inflammatory and neuropathic pain.

Topics: Analgesics; Animals; Behavior, Animal; Carrageenan; Female; Hyperalgesia; Inflammation; Male; Mice; Morphinans; Neuralgia; Nociception; Rats; Sciatic Nerve; Spinal Cord Injuries

Chronic inflammation mediated by microglial cells is the fundamental process contributing to the death of dopamine (DA)-producing neurons in the brain. Production of inflammatory products by these microglial cells characterizes the slow destructive process in Parkinson's disease (PD). The activation of microglial cells and the generation of pro-inflammatory cytokines that characterize PD are mediated by several different signaling pathways, with the activation of the respiratory burst by microglial cells being a critical event in the ultimate toxicity of DA-neurons. The work on our lab is concerned with understanding the mechanisms of activation, response, and therapeutic targets of microglial cells, with the aim to provide more effective treatments for PD and other inflammatory diseases of the CNS.

Topics: Brain; Chronic Disease; Cytokines; Dopamine; Humans; Inflammation; Microglia; Morphinans; Parkinson Disease

Dimemorfan (a sigma1 receptor agonist) showed neuroprotective properties in animal models of inflammation-mediated neurodegenerative conditions, but its effects on inflammatory cells and systemic inflammation remain unclear.. The effects of dimemorfan on phorbol-12-myristate-13-acetate (PMA)- and N-formyl-methionyl-leucyl-phenylalanine (fMLP)- induced neutrophils and lipopolysaccharide (LPS)-activated microglial cells, as well as LPS-induced endotoxin shock in mice were elucidated.. Dimemorfan decreased PMA- and fMLP-induced production of reactive oxygen species (ROS) and CD11b expression in neutrophils, through mechanisms independent of sigma1 receptors, possibly by blocking ROS production and G-protein-mediated intracellular calcium increase. Dimemorfan also inhibited LPS-induced ROS and nitric oxide (NO) production, as well as that of monocyte chemoattractant protein-1 and tumour necrosis factor-alpha (TNF-alpha), by inhibition of NADPH oxidase (NOX) activity and suppression of iNOS up-regulation through interfering with nuclear factor kappa-B (NF-kappaB) signalling in microglial cells. Treatment in vivo with dimemorfan (1 and 5 mg kg(-1), i.p., at three successive times after LPS) decreased plasma TNF-alpha, and neutrophil infiltration and oxidative stress in the lung and liver.. Our results suggest that dimemorfan acts via sigma1 receptor-independent mechanisms to modulate intracellular calcium increase, NOX activity, and NF-kappaB signalling, resulting in inhibition of iNOS expression and NO production, and production of pro-inflammatory cytokines. These effects may contribute its anti-inflammatory action and protective effects against endotoxin shock in mice.

Topics: Animals; Anti-Inflammatory Agents; Blotting, Western; Calcium; Cytokines; Fluorescent Antibody Technique; Humans; I-kappa B Proteins; Inflammation; Lipopolysaccharides; Macrophage-1 Antigen; Mice; Morphinans; N-Formylmethionine Leucyl-Phenylalanine; NADPH Oxidases; Neutrophils; Nitric Oxide; Nitric Oxide Synthase Type II; Reactive Oxygen Species; Shock, Septic; Tetradecanoylphorbol Acetate; Transcription Factor RelA; Up-Regulation

Opioids induce analgesia by activating opioid receptors not only within the central nervous system but also on peripheral sensory neurons. This study investigated peripherally mediated antinociception produced by the mu-opioid receptor agonist 2-[(4,5alpha-epoxy-3-hydroxy-14beta-methoxy-17-methylmorphinan-6beta-yl)amino]acetic acid (HS-731) after s.c. and oral administration in rats with carrageenan-induced hindpaw inflammation. Antinociceptive effects after s.c. administration were assessed 3 h after intraplantar carrageenan injection and compared with those of centrally acting mu-opioid agonists 14-methoxymetopon and morphine. Opioid agonists caused dose-dependent increases in inflamed paw withdrawal latencies to mechanical and thermal stimulation. The time course of action was different, in that HS-731 (20 microg/kg s.c.) produced significant long-lasting effects up to 4 h after administration, whereas 14-methoxymetopon (20 microg/kg) and morphine (2 mg/kg) reached their peak of action at 10 to 30 min, and their effect declined rapidly thereafter. Subcutaneous administration of the peripherally selective opioid antagonist naloxone methiodide inhibited antinociception elicited by HS-731 (20 microg/kg s.c.), whereas it was ineffective against 14-methoxymetopon (20 microg/kg s.c.). Moreover, the antinociception produced by 100 microg/kg s.c. HS-731 was dose-dependently reversed by s.c. naloxone methiodide. This indicates that HS-731 preferentially activates peripheral opioid receptors, whereas 14-methoxymetopon mediates analgesia via central mechanisms. Orally administered HS-731 significantly reduced hyperalgesia in the inflamed paw induced by carrageenan, which was reversible by s.c. administered naloxone methiodide. These results show that systemic (s.c. and oral) treatment with the mu-opioid agonist HS-731 produces potent and long-lasting antinociception through peripheral mechanisms in rats with carrageenan-induced hindpaw inflammation.

Topics: Administration, Oral; Analgesics, Opioid; Animals; Carrageenan; Dose-Response Relationship, Drug; Epoxy Compounds; Hindlimb; Inflammation; Injections, Subcutaneous; Male; Morphinans; Pain; Rats; Rats, Sprague-Dawley; Receptors, Opioid, mu

Eighteen sinomenine derivatives on ring C were prepared, and their anti-inflammatory activities were also investigated. Most of these derivatives showed mild to moderate activities. Compounds 4a, 4c and 5b showed better anti-inflammatory activity. So further modification of the ring C in sinomenine should be worthwhile.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Female; Inflammation; Male; Mice; Molecular Structure; Morphinans

Due to low central nervous system (CNS) bioavailability of delta-opioid peptides, little is known about the effect of systemic administration of delta-opioid receptor ligands. The present study examined the effect of non-peptidergic delta-opioid receptor agonists, (+)-4-[(alphaR)-alpha-((2R,5R)-4-Allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide (SNC80) and (-)dibenzoyl-L-tartaric acid salt (SNC86), on the activity of alpha-motoneurons in decerebrate-spinal rats. The flexor reflex was facilitated by C-afferent conditioning inputs, shown by a decrease in mechanical threshold and increase in touch- and pinch-evoked responses. Systemic administration of SNC80 (10 micromol/kg) prevented and reversed the neuronal hyperactivity. We further examined the effect of this agonist on the hypersensitivity of the flexor reflex induced by intraplantar injection of Freund's adjuvant. SNC80 dose-dependently (1, 3, 5 and 10 micromol/kg) increased the mechanical threshold and decreased touch-, pinch- and Abeta-afferent inputs-evoked responses. Similar effects were seen with SNC86 (5 micromol/kg). Pretreatment with either naloxone (20 micromol/kg, i.p.) or (Cyclopropylmethyl)-6,7-dehydro-4,5alpha-epoxy-14beta-ethoxy-5beta-methylindolo [2',3':6',7']morphinan-3-ol hydrochloride (SH378; 5 micromol/kg, intraarterially (i.a.)), a novel selective delta-opioid receptor antagonist, completely abolished the anti-hypersensitivity effect of SNC80. The effect of SNC80 remained following intrathecal administration of mu-opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP; 1.5 nmol). These results indicate that systemic injection of SNC80 exerted antihypersensitivity in models of both acute and tonic nociception and these effects are mediated mainly through a spinal delta-opioid mechanism.

Topics: Afferent Pathways; Animals; Benzamides; Central Nervous System; Conditioning, Psychological; Dose-Response Relationship, Drug; Drug Hypersensitivity; Electric Stimulation; Electrophysiology; Freund's Adjuvant; Hindlimb; Indoles; Inflammation; Male; Morphinans; Motor Neurons; Naloxone; Narcotic Antagonists; Pain Measurement; Piperazines; Rats; Rats, Wistar; Receptors, Opioid, delta; Receptors, Opioid, mu; Reflex; Somatostatin; Tartrates; Touch