phenanthrenes and Inflammation

Covering: 2000 to 2020 Triptolide is a bioactive diterpene triepoxide isolated from Tripterygium wilfordii Hook F, a traditional Chinese medicinal plant whose extracts have been used as anti-inflammatory and immunosuppressive remedies for centuries. Although triptolide and its analogs exhibit potent bioactivities against various cancers, and inflammatory and autoimmune diseases, none of them has been approved to be used in the clinic. This review highlights advances in material sourcing, molecular mechanisms, clinical progress and new drug design strategies for triptolide over the past two decades, along with some prospects for the future course of development of triptolide.

Topics: Animals; Autoimmune Diseases; Diterpenes; Drug Design; Drug Discovery; Epoxy Compounds; Forecasting; Humans; Inflammation; Neoplasms; Phenanthrenes; Tripterygium

As a natural quinone compound, the medicinal value of cryptotanshinone (CT) has received increasing attentions, but there is no systematic literature review that describes the pharmacological activity of CT. This paper reviewed the pharmacology researches of CT, with a primary focus on its anti-tumor activity. We also discussed the underlying molecular mechanisms, and proposed future outlooks. In addition to anti-tumor activity, CT was found to have anti-inflammatory, neuroprotective, cardioprotective, visceral protective, anti-metabolic disorders and other abilities. Furthermore, the potential molecular mechanisms contributing to the anti-tumor effect of CT likely involve the following aspects: the induction of apoptosis, targeting of ER and AR, reversion of MDR, combined pharmacotherapy, and the inhibition of cell proliferation, migration, and invasion. We also found that different pharmacological effects involved various signaling pathways. Among them, STAT3-related signaling pathways played a vital role in the CT-mediated induction of tumor cell apoptosis and proliferation, while NF-κB signal pathway also was essential for inhibition of inflammation by CT. Furthermore, CT could significantly enhance the activities of several anticancer drugs and reverse their resistances in tumors. Therefore, we proposed suggestions for future studies of CT, including enhancing anti-tumor activity by targeting STAT3-related receptors, targeting NF-κB-related pathways to inhibit inflammatory responses, enhancing anti-tumor efficacy by combining with anti-tumor drugs, and further studying the dose-effect relationship to ensure safer and more effective applications of CT.

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Drug Resistance, Neoplasm; Humans; Inflammation; NF-kappa B; Phenanthrenes; Protective Agents; Signal Transduction; STAT3 Transcription Factor

Apart from cancer chronic (auto)immune-mediated diseases are a major threat for patients and a challenge for physicians. These conditions include classic autoimmune diseases like systemic lupus erythematosus, systemic sclerosis and dermatomyositis and also immune-mediated inflammatory diseases such as rheumatoid arthritis and psoriasis. Traditional therapies for these conditions include unspecific immunosuppressants including steroids and cyclophosphamide, more specific compounds such as ciclosporin or other drugs which are thought to act as immunomodulators (fumarates and intravenous immunoglobulins). With increasing knowledge about the underlying pathomechanisms of the diseases, targeted biologic therapies mainly consisting of anti-cytokine or anti-cytokine receptor agents have been developed. The latter have led to a substantial improvement of the induction of long term remission but drug costs are high and are not affordable in all countries. In China an extract of the herb Tripterygium wilfordii Hook F. (TwHF) is frequently used to treat autoimmune and/or inflammatory diseases due to its favourable cost-benefit ratio. Triptolide has turned out to be the active substance of TwHF extracts and has been shown to exert potent anti-inflammatory and immunosuppressive effects in vitro and in vivo. There is increasing evidence for an immunomodulatory and partly immunosuppressive mechanism of action of triptolide. Thus, compounds such as triptolide or triptolide derivatives may have the potential to be developed as a new class of drugs for these diseases. In this review we summarize the published knowledge regarding clinical use, pharmacokinetics and the possible mode of action of triptolide in the treatment of inflammatory diseases with a particular focus on psoriasis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Autoimmune Diseases; Diterpenes; Epoxy Compounds; Humans; Immunosuppressive Agents; Inflammation; Phenanthrenes; Psoriasis; Tripterygium

Parkinson's disease (PD) is a degenerative neurological disorder characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) of the brain. The loss of the dopaminergic projection from the SNpc deprives the striatum of dopamine and results in a myriad of motor signs, including tremor, rigidity and ataxia. Although the stimulus for the initiation of the degenerative process is not understood, 80% of the dopaminergic neurons in the SNpc must be lost before the clinical symptoms of the disease are observed. This suggests that the degenerative process is initiated many years before clinical presentation of the disease. The neurodegeneration observed in PD is accompanied by inflammatory processes, and it has been suggested that anti-inflammatory drugs may be useful in slowing disease progression once the clinical signs of PD have been observed. This review summarizes and evaluates the progress that has been made in this area of research since 2006.

Topics: Animals; Anti-Inflammatory Agents; Benzoates; Bridged-Ring Compounds; Diterpenes; Drug Evaluation, Preclinical; Glucosides; Humans; Inflammation; Monoterpenes; Parkinson Disease; Phenanthrenes; Purines

Topics: Abietanes; Animals; Anti-Bacterial Agents; Bacteria; Bacterial Infections; Bandages; Diterpenes; Female; Humans; In Vitro Techniques; Inflammation; Male; Microbial Sensitivity Tests; Phenanthrenes; Rats; Resins, Plant; Tars; Wound Healing; Wound Infection; Zinc; Zinc Oxide

The rhizome of Dioscorea batatas Decne (called Chinses yam) widely distributed in East Asian countries including China, Japan, Korea and Taiwan has long been used in oriental folk medicine owing to its tonic, antitussive, expectorant and anti-ulcerative effects. It has been reported to have anti-inflammatory, antioxidative, cholesterol-lowering, anticholinesterase, growth hormone-releasing, antifungal and immune cell-stimulating activities.. Neuroinflammation caused by activated microglia contributes to neuronal dysfunction and neurodegeneration. In the present study, the anti-neuroinflammatory activity of 6,7-dihydroxy-2,4-dimethoxy phenanthrene (DHDMP), a phenanthrene compound isolated from Dioscorea batatas Decne, was examined in microglial and neuronal cells.. A natural phenanthrene compound, DHDMP, was isolated from the peel of Dioscorea batatas Decne. The anti-neuroinflammatory capability of the compound was examined using the co-culture system of BV2 murine microglial and HT22 murine neuronal cell lines. The expression levels of inflammatory mediators and cytoprotective proteins in the cells were quantified by enzyme-linked immunosorbent assay and Western blot analysis.. DHDMP at the concentrations of ≤1 μg/mL did not exhibit a cytotoxic effect for BV2 and HT22 cells. Rather DHDMP effectively restored the growth rate of HT22 cells, which was reduced by co-culture with lipopolysaccharide (LPS)-treated BV2 cells. DHDMP significantly decreased the production of proinflammatory mediators, such as nitric oxide, tumor necrosis factor-α, interleukin-6, inducible nitric oxide synthase, and cyclooxygenase-2 in BV2 cells. Moreover, DHDMP strongly inhibited the nuclear translocation of nuclear factor κB (NF-κB) and phosphorylation of p38 mitogen-activated protein kinase (MAPK) in BV2 cells. The compound did not affect the levels and phosphorylation of ERK and JNK. Concurrently, DHDMP increased the expression of heme oxygenase-1 (HO-1), an inducible cytoprotective enzyme, in HT22 cells.. Our findings indicate that DHDMP effectively dampened LPS-mediated inflammatory responses in BV2 microglial cells by suppressing transcriptional activity of NF-κB and its downstream mediators and contributed to HT22 neuronal cell survival. This study provides insight into the therapeutic potential of DHDMP for inflammation-related neurological diseases.

Topics: Animals; Dioscorea; Gene Expression Regulation; Humans; Inflammation; Microglia; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Rats

Inflammation is a biological process that exists in a large number of diseases. NF-κB has been proven to play a pivotal role in the development of inflammation. New drugs aimed at inhibiting the expression of NF-κB have gained attention from researchers. Sirt1 has an anti-inflammatory function, and the circRNA encoded by the Sirt1 gene may also play roles in the anti-inflammatory reaction of Sirt1. In the present study, LPS-treated RAW264.7 cells were used as an inflammatory cell model, and tanshinone IIA sodium sulfonate (TSS) was used as a therapeutic drug. We found that TSS downregulated LPS-induced TNF-α and IL-1β expression nearly threefold. LPS reduced Circ-sirt1 mRNA expression by one-third, while TSS started this phenomenon. In addition, overexpression/knockdown of Circ-sirt1 neutralized the function of TSS by regulating the translocation of NF-κB. Thus, we proved that TSS has an anti-inflammatory function by upregulating circ-Sirt1 and subsequently inhibiting the translocation of NF-κB. An in vivo experiment was also performed to confirm the protective function of TSS on inflammation. These results indicated that TSS is a potential treatment for inflammation.

Topics: Animals; Anti-Inflammatory Agents; Drugs, Chinese Herbal; Gene Expression Regulation; Inflammation; Mice; NF-kappa B; Phenanthrenes; RAW 264.7 Cells; RNA, Circular; Salvia miltiorrhiza; Sirtuin 1; Tumor Necrosis Factor-alpha; Up-Regulation

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

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; Apoptosis; Female; Ferroptosis; Humans; Inflammation; Matrix Metalloproteinases; Membrane Potential, Mitochondrial; Oxidative Stress; Phenanthrenes; Polycystic Ovary Syndrome; Rats

Diabetic nephropathy (DN) is a complication of diabetes. This study sought to explore the mechanism of triptolide (TP) in podocyte injury in DN. DN mice were induced by high-fat diet&streptozocin and treated with TP. Fasting blood glucose, 24 h urine microalbumin (UMA), the pathological changes of renal tissues, and ultrastructure of renal podocytes were observed. Podocytes (MPC5) were induced by high-glucose (HG)

Topics: Animals; Apoptosis; Brain-Derived Neurotrophic Factor; Diabetes Mellitus; Diabetic Nephropathies; Diterpenes; Epoxy Compounds; Glucose; Inflammation; Irbesartan; Mice; MicroRNAs; Oxidative Stress; Phenanthrenes; Podocytes

Delayed-onset muscle soreness (DOMS) is associated with exercise-induced muscle damage and inflammation, which is mainly caused by prolonged eccentric exercise in humans. Triptolide, an extract from the Chinese herb Tripterygium wilfordii Hook F, has been used for treating autoimmune and inflammatory diseases in clinical practice. However, whether triptolide attenuates acute muscle damage is still unclear. Here, we examined the effect of triptolide on carrageenan-induced DOMS in rats. Rats were injected with 3% of carrageenan into their muscles to induce acute left gastrocnemius muscular damage, and triptolide treatment attenuated carrageenan-induced acute muscular damage without affecting hepatic function. Triptolide can significantly decrease lipid hydroperoxide and nitric oxide (NO) levels, proinflammatory cytokine production, and the activation of nuclear factor (NF)-ĸB, as well as increase a reduced form of glutathione levels in carrageenan-treated rat muscles. At the enzyme levels, triptolide reduced the inducible nitric oxide synthase (iNOS) expression and muscular myeloperoxidase (MPO) activity in carrageenan-treated DOMS rats. In conclusion, we show that triptolide can attenuate muscular damage by inhibiting muscular oxidative stress and inflammation in a carrageenan-induced rat DOMS model.

Topics: Animals; Carrageenan; Epoxy Compounds; Humans; Inflammation; Models, Animal; Myalgia; Myositis; Oxidative Stress; Phenanthrenes; Rats

Triptolide is naturally isolated from Tripterygium wilfordii Hook F., possessing multiple biological activities. Hepatotoxicity is one of the main side effects of triptolide. However, the effect of triptolide on nonalcoholic fatty liver disease remains unknown (NAFLD).. This study aimed to observe the amelioration of triptolide against NAFLD and investigate the engaged mechanism.. Two typical animal models of NAFLD, obese db/db mice and methionine/choline-deficient (MCD) diet-fed mice, were used. Hepatic steatosis, inflammation, and fibrosis were evaluated by H&E and Masson staining. Oil red O staining and lipid extraction analysis were used to detect fat content in mice livers. Expression of lipid metabolism, inflammatory and fibrogenic genes was also detected by Real-time PCR and Western blotting, respectively. Phosphoproteomics, molecular docking, and TR-FRET assay were performed to provide further insight into how triptolide improved NAFLD.. This study demonstrates that dose-related triptolide as an allosteric AMPK agonist has the potential to alleviate NAFLD without hepatotoxicity.

Topics: AMP-Activated Protein Kinases; Animals; Diterpenes; Epoxy Compounds; Inflammation; Lipid Metabolism; Lipids; Liver; Liver Cirrhosis; Mice; Mice, Inbred C57BL; Molecular Docking Simulation; Non-alcoholic Fatty Liver Disease; Phenanthrenes

Triptolide (TP), the main active compound extracted from medicine-

Topics: Animals; Cytochrome P-450 Enzyme System; Diterpenes; Enzyme Induction; Epoxy Compounds; Inflammation; Liver; Male; Mice; Mice, Inbred C57BL; Oxidative Stress; Phenanthrenes

Streptococcus suis is a highly zoonotic pathogen that is a serious threat to human health and the development of the pig industry worldwide. The virulence factors produced during S. suis infection play an important role, and the pore-forming activity of suilysin is considered an important virulence-related factor, especially in meningitis. Treatment of S. suis infection with traditional antibiotics is becoming increasingly challenging due to bacterial resistance. The purpose of this study is to verify the role of cryptotanshinone in the process of S. suis infection and provide a new drug precursor for the treatment of S. suis infection.. In this study, we used circular dichroism spectroscopy to demonstrate that cryptotanshinone alters the secondary structure of suilysin. The results of the antibacterial activity and haemolysis assays showed cryptotanshinone could inhibit the pore-forming activity of suilysin without affecting bacterial growth or its expression. We also showed that cryptotanshinone reduces bacterial damage and penetration in vitro, reduce the S. suis-induced inflammatory response and provide protection against bacterial infections in vivo and in vitro.. Cryptotanshinone is a potential compound precursor for treating S. suis infection.. Cryptotanshinone may be a promising leading compound for S. suis infection and related diseases.

Topics: Animals; Anti-Bacterial Agents; Hemolysin Proteins; Hemolysis; Humans; Inflammation; Mice; Phenanthrenes; Protein Structure, Secondary; Streptococcal Infections; Streptococcus suis; Virulence; Virulence Factors

This study investigated the neuroprotective effects of triptolide (TPL) in a rat model of cardiopulmonary bypass with deep hypothermia circulatory arrest (DHCA). Rats were randomly divided into six groups: control, sham, DHCA, and DHCA + TPL (100, 200, 300 μg/kg). Neurobehavioral functions were measured using the elevated plus-maze, Y-maze, and Morris water maze tests. Levels of inflammatory cytokines, oxidative stress indices, and brain neurotrophins were measured by ELISA. Microglial activation and cell death was measured by immunofluorescence staining and TUNEL assay, respectively. Finally, activation of the Nrf2 pathway and NF-κB were detected by western blot. The elevated plus-maze, Y-maze, and Morris water maze tests all showed that TPL mitigated anxiety-like behavior, working memory, spatial learning, and memory in DHCA rats. TPL inhibited inflammatory responses and oxidative stress, as well as increased brain neurotrophin levels in DHCA rats. Moreover, TPL attenuated microglia activation and cell death in DHCA rats. Finally, TPL activated the Nrf2 pathway and inhibited NF-κB activity in DHCA rats. These results demonstrated that TPL improved neurobehavioral functions, neuroinflammation, and oxidative stress in DHCA rats, which may be associated with the Nrf2 and NF-κB pathways.

Topics: Animals; Behavior, Animal; Brain; Cell Death; Circulatory Arrest, Deep Hypothermia Induced; Diterpenes; Epoxy Compounds; Inflammation; Male; Memory, Short-Term; Microglia; Nerve Growth Factors; Neuroprotective Agents; Oxidative Stress; Phenanthrenes; Rats; Spatial Learning

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

Rheumatoid arthritis (RA), recognized as a common chronic autoimmune disease, is characterized by the excessive proliferation and inflammatory infiltration of fibroblast-like synoviocytes (FLS). In this study, our purpose is to elucidate the mechanisms of triptolide (TPL) in the treatment of RA by regulating the long non-coding RNA (lncRNA) ENST00000619282, which promoted apoptosis and reduced inflammatory infiltration of FLS in RA (RA-FLS). RA-FLS was treated with different concentrations of TPL at different time points. CCK-8 assay, ELISA, RT-qPCR, immunofluorescence, TUNEL assay, and the transmission electron microscopy were used to measure the changes of cell viability, apoptosis, and the release of inflammatory cytokines. Next, the involvement of ENST00000619282 in TPL-mediated protection against RA was explored. ENST00000619282 expression was significantly increased in the peripheral blood mononuclear cells (PBMCs) of RA patients. ENST0000061928 expression in RA PBMCs was positively associated with ESR, RF, CCP, and DAS28, while TPL treatment led to a downregulation of ENST00000619282. In addition, ENST00000619282 was significantly increased in RA-FLS. Furthermore, overexpression of ENST00000619282 elevated the levels of pro-apoptotic and pro-inflammatory factors, while reduced the levels of anti-apoptotic proteins and antiinflammatory factors. Besides, TPL treatment could reverse these effects by ENST00000619282 overexpression. The anti-RA potential of TPL might be achieved by downregulating ENST00000619282, thereby promoting apoptosis, and reducing the inflammatory response in RA.

Topics: Apoptosis; Arthritis, Rheumatoid; Cell Proliferation; Cells, Cultured; Diterpenes; Epoxy Compounds; Fibroblasts; Humans; Inflammation; Leukocytes, Mononuclear; Phenanthrenes; RNA, Long Noncoding; Synoviocytes

This study is to determine the role and mechanism of cryptotanshinone (CTS) in allergic airway inflammation. Asthma induced by OVA was established in BALB/c mice. We found increased airway hyperresponsiveness (AHR), increased inflammatory cell infiltration, elevated levels of TNF-α, interleukin-1β (IL-1β), IL-4, IL-5, IL-6 and IL-13, decreased interferon gamma (IFN-γ) in lung tissue, increased content of total immunoglobulin E (IgE), OVA specific IgE, Eotaxin, ICAM-1, VCAM-1, nuclear factor-kappaB (NF-κB) and phosphorylation of p38 MAPK in lung tissue. However, the administration of CTS significantly decreased AHR in asthmatic mice, reduced inflammation around the bronchioles and inflammatory cells around airway, regulated cytokine production, reduced the total IgE and OVA-specific IgE levels, and inhibited NF-κB activation and p38 MAPK phosphorylation. In vitro experiments in 16 HBE cells revealed that CTS attenuated CAM-1 and IL-6 expression. These results indicate that CTS alleviates allergic airway inflammation by modulating p38 MAPK phosphorylation and NF-κB activation.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemotaxis, Leukocyte; Cytokines; Drugs, Chinese Herbal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Phosphorylation

Salvia miltiorrhiza, known as Danshen in Chinese, has been widely used to treat cardiovascular diseases in China. Tanshinone I (Tan I) and cryptotanshinone (CST) are the lipid-soluble and effective components from Salvia miltiorrhiza. However, the molecular mechanism of Tan I and CST for treating inflammation is still not known. Therefore, this study was designed to use network pharmacology-based strategy to predict therapeutic targets of Tan I and CST against inflammation, and further to investigate the pharmacological molecular mechanism in vitro. Inflammation targets were identified and followed by acquisition of verified targets of Tan I and CST. After constructing target-functional protein interaction network of Tan I and CST against inflammation, the core therapeutic targets of Tan I and CST against inflammation were obtained. Further, pathway enrichment analyses were performed on core therapeutic targets to evaluate key signaling pathways of Tan I and CST against inflammation. As revealed in network pharmacology analysis, 8 key hub targets for Tan I and CST against inflammation were identified, respectively: JUN, VEGFA, IL-6, TNF, MAPK8, CXCL8, and PTGS2 for Tan I, while STAT3, AKT1, CCND1, MAPK14, VEGFA, ESR1, MAPK8 and AR for CST. Pathway enrichment analysis by DAVID database indicated that Tan I and CST principally regulated the inflammation-associated pathway, such as TLR, JAK-STAT signaling pathway, focal adhesion, apoptosis, mTOR signaling pathway. In vitro, we found that both Tan I and CST exerts significantly effect on LPS stimulated NO secretion and iNOS expression in macrophages. Taken together, our data elucidate that anti-inflammatory pharmacological activities of Tan I and CST may be predominantly related to inhibition of TLR signaling pathway and regulating iNOS synthesis. These findings highlight the predicted therapeutic targets may be potential targets of Tan I and CST for anti-inflammation treatment.

Topics: Abietanes; Animals; Anti-Inflammatory Agents; Drugs, Chinese Herbal; Humans; Inflammation; Lipopolysaccharides; Macrophage Activation; Mice; Nitric Oxide; Nitric Oxide Synthase Type II; Phenanthrenes; Protein Interaction Mapping; Protein Interaction Maps; RAW 264.7 Cells; Salvia miltiorrhiza; Signal Transduction; Toll-Like Receptors

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

In general, anti-inflammatory treatment is considered for multiple liver diseases despite the etiology. But current drugs for alleviating liver inflammation have defects, making it necessary to develop more potent and safer drugs for liver injury. In this study, we screened a series of (dihydro-)stilbene or (dihydro-)phenanthrene derivatives extracted from Pholidota chinensis for their potential biological activities. Among 31 compounds, the dihydro-stilbene gigantol exerted most potent protective effects on human hepatocytes against lithocholic acid toxicity, and exhibited solid antioxidative and anti-inflammatory effect in vitro. In mice with CCl

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Antioxidants; Bibenzyls; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Complement Membrane Attack Complex; Guaiacol; Hepatocytes; Humans; Inflammation; Lipid Peroxidation; Lithocholic Acid; Liver; Male; Mice, Inbred ICR; Oxidative Stress; Phenanthrenes; Proteome; Rats, Sprague-Dawley; Stilbenes

Doxorubicin is effective in a variety of solid and hematological malignancies. Unfortunately, clinical application of doxorubicin is limited due to a cumulative dose-dependent cardiotoxicity. Dihydrotanshinone I (DHT) is a natural product from Salvia miltiorrhiza Bunge with multiple anti-tumor activity and anti-inflammation effects. However, its anti-doxorubicin-induced cardiotoxicity (DIC) effect, either in vivo or in vitro, has not been elucidated yet. This study aims to explore the anti-inflammation effects of DHT against DIC, and to elucidate the potential regulatory mechanism.. Effects of DHT on DIC were assessed in zebrafish, C57BL/6 mice and H9C2 cardiomyocytes. Echocardiography, histological examination, flow cytometry, immunochemistry and immunofluorescence were utilized to evaluate cardio-protective effects and anti-inflammation effects. mTOR agonist and lentivirus vector carrying GFP-TFEB were applied to explore the regulatory signaling pathway.. DHT improved cardiac function via inhibiting the activation of M1 macrophages and the excessive release of pro-inflammatory cytokines both in vivo and in vitro. The activation and nuclear localization of NF-κB were suppressed by DHT, and the effect was abolished by mTOR agonist with concomitant reduced expression of nuclear TFEB. Furthermore, reduced expression of nuclear TFEB is accompanied by up-regulated phosphorylation of IKKα/β and NF-κB, while TFEB overexpression reversed these changes. Intriguingly, DHT could upregulate nuclear expression of TFEB and reduce expressions of p-IKKα/β and p-NF-κB.. Our results demonstrated that DHT can be applied as a novel cardioprotective compound in the anti-inflammation management of DIC via mTOR-TFEB-NF-κB signaling pathway. The current study implicates TFEB-IKK-NF-κB signaling axis as a previously undescribed, druggable pathway for DIC.

Topics: Animals; Antibiotics, Antineoplastic; Apoptosis; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Cardiotoxicity; Cell Proliferation; Cells, Cultured; Cytokines; Doxorubicin; Furans; Gene Expression Regulation; Humans; Inflammation; Macrophages; Male; Mice; Mice, Inbred C57BL; Myocytes, Cardiac; NF-kappa B; Phenanthrenes; Phosphorylation; Quinones; Zebrafish

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

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

Cryptotanshinone (CT) is a diterpene quinone compound from. This study evaluates the wound healing activity of CT by employing an excisional wound splinting model in db/db mice.. Wounds were induced at the dorsum of non-diabetic (db/+) and diabetic (db/db) mice and treated with sodium carboxymethyl cellulose (CMC-Na) or 300 mg/kg/d CT for 16 days. Wound closure was measured every two days. Body weight, fasting blood glucose, re-epithelialization, granulation, leukocyte infiltration, capillary density, collagen deposition and expressions of CXCL1, CXCL2, VEGF, Ang-1, p-eNOS, eNOS, α-SMA, MMP2 and MMP9 were analysed. Expression of VEGF and tube formation was measured. CT significantly accelerated rate of wound closure, as the contraction ratio increased from 68% (non-treated group) to 83% (CT-treated group) at days 16 post-injury. A significant increase was observed in re-epithelialization and granulation tissue formation. Mechanistically, CT suppressed leukocyte infiltration and CXCL1 and CXCL2 expression. CT treatment also increased blood vessel density and expression level of VEGF, Ang-1 and p-eNOS.. Our study provides evidence that CT could be developed as a potential therapeutic agent for the treatment of chronic diabetic wound healing.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; Extracellular Matrix; Fibroblasts; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Male; Mice; Neovascularization, Physiologic; Phenanthrenes; Time Factors; Wound Healing

Chemotherapy drugs exerts beneficial antitumor activity before and after cancer surgery. Post-injury complications are a potential hazard after surgical tumor resection. Inflammation caused by surgical stress is known to promote the progression of post-injury complications. Recent studies have found that chemotherapy drugs can promote post-injury inflammatory response, leading to increased post-injury complications. Imidazole derivatives have effective anticancer activity. However, the impact of post-operative inflammation caused by imidazole derivatives is unclear. In this study, two novel phenanthroimidazole derivatives (L082 and L142) were synthesized and characterized. These compounds showed significant inhibitory effects on different tumor cells. The compound L082 also inhibited liver cancer in vivo. In addition, L082 played a significant role in inhibiting the accumulation of inflammatory cells and promoting the elimination of inflammatory cells at the incision, which may be related to inhibiting the production of ROS and NO in oxidative and nitric stress. These results suggest that L082 can be used as a bifunctional drug to suppress tumors and reduce post-injury inflammation complications.

Topics: A549 Cells; Animals; Animals, Genetically Modified; Anti-Inflammatory Agents; Antineoplastic Agents; Dose-Response Relationship, Drug; HeLa Cells; Hep G2 Cells; Humans; Inflammation; Liver Neoplasms, Experimental; Oxidative Stress; Phenanthrenes; Zebrafish

Endoplasmic reticulum stress is a cellular phenomenon that has been associated with metabolic disorders, contributing to the development of obesity, fatty liver disease, and dyslipidemias. Under metabolic overload conditions, in cells with a high protein-secretory activity, such as hepatocytes and Langerhans β cells, the unfolded protein response (UPR) is critical in to maintain protein homeostasis (proteostasis). UPR integrated by a tripartite signaling system, through activating transcription factor 6, protein kinase R-like endoplasmic reticulum kinase (PERK), and inositol-requiring enzyme 1, regulates gene transcription and translation to resolve stress and conserve proteostasis. In the current study, we demonstrated in hepatocytes under metabolic overload by saturated palmitic and stearic fatty acids, through activation of PERK signaling and CCAAT-enhancer-binding protein homologous protein (CHOP) transcription factor, an association with the expression of cyclooxygenase 2. More important, isolated exosomes from supernatants of macrophages exposed to lipopolysaccharides can also induce a metainflammation phenomenon, and when treated on hepatocytes, induced a rearrangement in cholesterol metabolism through sterol regulatory element-binding protein 2 (SREBP2), low-density lipoprotein receptor (LDLR), apolipoprotein A-I, and ABCA1. Moreover, we demonstrate the cellular effect of terpene-derived molecules, such as cryptotanshinone, isolated of plant Salvia brandegeei, regulating metainflammatory conditions through PERK pathway in both hepatocytes and β cells. Our data suggest the presence of a modulatory mechanism on specific protein translation process. This effect could be mediated by eukaryotic initiation factor-4A, evaluating salubrinal as a control molecule. Likewise, the protective mechanisms of unsaturated fatty acids, such as oleic and palmitoleic acid were confirmed. Therefore, modulation of metainflammation suggests a new target through PERK signaling in cells with a high secretory activity, and possibly the regulation of cholesterol in hepatocytes is promoted via exosomes.

Topics: Animals; Camphanes; Cholesterol; Cyclooxygenase 2; Drugs, Chinese Herbal; eIF-2 Kinase; Endoplasmic Reticulum Stress; Exosomes; Fatty Acids; Hepatocytes; Inflammation; Insulin-Secreting Cells; Mice; Panax notoginseng; Phenanthrenes; Protein Biosynthesis; Rats; RAW 264.7 Cells; Salvia miltiorrhiza; Signal Transduction; Transcription Factor CHOP; Tunicamycin

Non-alcoholic fatty liver disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited lipid accumulation in oleic acid (OA) and palmitic acid (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed lipogenesis by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced inflammation in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased lipid accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.

Topics: Animals; Dose-Response Relationship, Drug; Hep G2 Cells; Humans; Inflammation; Lipogenesis; Male; Mice; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease; Phenanthrenes; Random Allocation

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

Sodium tanshinone IIA sulfonate (STS), a water-soluble derivative of tanshinone IIA, has been demonstrated to have potent anti-inflammatory properties. However, the protective effects of STS on lipopolysaccharide (LPS)-induced inflammation in endothelial cells remain to be elucidated. In the present study, human umbilical vein endothelial cells (HUVECs) were used to explore the effects of STS on LPS-induced inflammation and the molecular mechanism involved. HUVECs were pretreated with STS for 2 h, followed by stimulation with LPS. Then expression and secretion of tumor necrosis factor (TNF)-α and interleukin (IL)-1β, and the activation of nuclear factor-κB (NF-κB) were assessed. The results demonstrated that STS significantly decreased LPS-induced TNF-α and IL-1β protein expression in HUVECs. Similarly, the increased levels of TNF-α and IL-1β in cell supernatants stimulated by LPS were also significantly inhibited by STS. Furthermore, STS inhibited LPS-induced NF-κB p65 phosphorylation and nuclear translocation. All the results suggest that STS prevents LPS-induced inflammation through suppressing NF-κB signaling pathway in endothelial cells, indicating the potential utility of STS for the treatment of inflammatory diseases.

Topics: Cell Survival; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Interleukin-1beta; Lipopolysaccharides; NF-kappa B; Nitriles; Phenanthrenes; Signal Transduction; Sulfones; Tumor Necrosis Factor-alpha

Inflammation may play an important role in the association between exposure to polycyclic aromatic hydrocarbons (PAHs) and atherosclerotic cardiovascular disease (ASCVD) risk. However, the underlying mechanisms remain unclear.. To investigate the association of PAHs exposure with ASCVD risk and effects of mean platelet volume (MPV) or Club cell secretory protein (CC16) on the association.. A total of 2022 subjects (689 men and 1333 women) were drawn from the baseline Wuhan residents of the Wuhan-Zhuhai Cohort study. Data on demography and the physical examination were obtained from each participant. Urinary monohydroxy PAH metabolites (OH-PAHs) levels were measured by a gas chromatography-mass spectrometry. We estimated the association between each OH-PAHs and the 10-year ASCVD risk or coronary heart disease (CHD) risk using logistic regression models, and further analyze the mediating effect of MPV or plasma CC16 on the association by using structural equation modeling.. The results of multiple logistic regression models showed that some OH-PAHs were positively associated with ASCVD risk but not CHD risk, including 2-hydroxyfluoren (β = 1.761; 95% CI: 1.194-2.597), 9-hydroxyfluoren (β = 1.470; 95% CI: 1.139-1.898), 1-hydroxyphenanthrene (β = 1.480; 95% CI: 1.008-2.175) and ΣOH-PAHs levels (β = 1.699; 95% CI: 1.151-2.507). The analysis of structural equation modeling shows that increased MPV and increased plasma CC16 levels contributed 13.6% and 15.1%, respectively, to the association between PAHs exposure and the 10-year ASCVD risk (p < 0.05).. Exposure to PAHs may increase the risk of atherosclerosis, which was partially mediated by MPV or CC16.

Topics: Cardiovascular Diseases; China; Cohort Studies; Environmental Exposure; Female; Gas Chromatography-Mass Spectrometry; Humans; Inflammation; Logistic Models; Male; Mean Platelet Volume; Middle Aged; Phenanthrenes; Polycyclic Aromatic Hydrocarbons

Donor-specific antibodies (DSAs) are major mediators of renal allograft injury, and strategies to inhibit DSAs are important in promoting long-term graft survival. Triptolide exhibits a wide spectrum of antiinflammatory and immunosuppressive activities, and in autoimmune diseases it inhibits autoantibody levels. In this study, we investigated the suppressive role of triptolide in the generation of DSAs in transplant recipients. We found that triptolide treatment of skin allograft recipients in mice significantly suppressed the development of circulating anti-donor-specific IgG and effectively alleviated DSA-mediated renal allograft injury, which led to prolonged allograft survival. In vitro studies revealed that triptolide inhibited the differentiation of B cells into CD138

Topics: Allografts; Animals; B-Lymphocytes; Diterpenes; Epoxy Compounds; Graft Rejection; Graft Survival; Immunosuppressive Agents; Inflammation; Isoantibodies; Kidney Transplantation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Phenanthrenes; Tissue Donors

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

Benefits from thrombolysis with recombinant tissue plasminogen activator (rt-PA) after ischemic stroke remain limited due to a narrow therapeutic window, low reperfusion rates, and increased risk of hemorrhagic transformations (HT). Experimental data showed that rt-PA enhances the post-ischemic activation of poly(ADP-ribose)polymerase (PARP) which in turn contributes to blood-brain barrier injury. The aim of the present study was to evaluate whether PJ34, a potent PARP inhibitor, improves poor reperfusion induced by delayed rt-PA administration, exerts vasculoprotective effects, and finally increases the therapeutic window of rt-PA. Stroke was induced by thrombin injection (0.75 UI in 1 μl) in the left middle cerebral artery (MCA) of male Swiss mice. Administration of rt-PA (0.9 mg kg

Topics: Animals; Edema; Endothelial Cells; Hemorrhage; Infarction, Middle Cerebral Artery; Inflammation; Male; Mice; Neuroprotective Agents; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Proteolysis; Recombinant Proteins; Reperfusion; Stroke; Thrombosis; Tissue Plasminogen Activator; Treatment Outcome; Vasospasm, Intracranial

Oxidative stress and inflammatory responses are closely implicated in the progression of renal interstitial fibrosis, thereby leading to chronic kidney disease. Cryptotanshinone (CTS) is a natural compound involved in antioxidant and anti-inflammatory activities. We evaluated the effects of CTS on inflammation and oxidative stress in obstructed kidneys. Mice received gastric gavage of CTS from 7 days before unilateral ureteral obstruction operation to 1 week after surgery. Administration of CTS at 50 and 100 mg/kg/day significantly decreased collagen production, as shown by Masson staining. Immunohistochemistry staining and RT-PCR confirmed that CTS reduced extracellular matrix proteins, such as fibronectin and collagen-1, in the obstructed kidneys in a dose-dependent manner. Furthermore, immunohistochemistry staining indicated that CTS inhibited infiltration of the macrophage (CD68-positive) and lymphocyte (CD3-positive) cells, which were associated with the suppression of the nuclear factor-κB signalling activation. CTS increased superoxide dismutase, catalase and glutathione while decreased malondialdehyde production. More importantly, CTS activated Nrf-2 and HO-1 in the obstructed kidneys for 7 days. CTS could protect renal interstitial fibrosis by ameliorating inflammation and oxidative stress, which might be through the regulation of NF-κB and Nrf-2/HO-1 signalling pathways.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Blotting, Western; Inflammation; Kidney; Mice; Mice, Inbred C57BL; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; Phenanthrenes; Real-Time Polymerase Chain Reaction; Ureteral Obstruction

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by cellular infiltration into the joints and cartilage destruction. Neutrophils play a crucial role in the pathogenesis of RA. Triptolide (TP) is a bioactive compound derived from Tripterygium wilfordii Hook F, which has been used in folk medicine as a treatment for a variety of inflammatory disorders, including RA, for many centuries. Previous studies have shown that TP possesses anti-arthritic activity. However, the anti-arthritic mechanism of TP remains to be fully defined. In the present study, we used the adjuvant-induced arthritis (AA) murine model of RA to investigate the impact of TP on RA and neutrophil function. TP alleviated AA by reducing neutrophil recruitment and suppressing the expression of interleukin-6 and tumour necrosis factor-α in vivo. TP also suppressed the expression of pro-inflammatory cytokines in neutrophils, promoted neutrophil apoptosis and inhibited the migration, NETosis and autophagy of neutrophils in vitro. Based on our findings, TP effectively ameliorates RA by down-regulating neutrophil inflammatory functions, indicating that TP represents a potential therapeutic agent for RA.

Topics: Animals; Apoptosis; Arthritis, Experimental; Arthritis, Rheumatoid; Autophagy; Chronic Disease; Cytokines; Diterpenes; Epoxy Compounds; Extracellular Traps; Inflammation; Leukocyte Elastase; Lipopolysaccharides; Male; Mice, Inbred C57BL; Neutrophil Infiltration; Neutrophils; Peroxidase; Phenanthrenes

Ionizing radiation-induced pulmonary injury is a major limitation of radiotherapy for thoracic tumors. We have demonstrated that triptolide (TPL) could alleviate IR-induced pneumonia and pulmonary fibrosis. In this study, we explored the underlying mechanism by which TPL mitigates the effects of radiotoxicity. The results showed that:(1) Alveolar macrophages (AMs) were the primary inflammatory cells infiltrating irradiated lung tissues and were maintained at a high level for at least 17 days, which TPL could reduce by inhibiting of the production of macrophage inflammatory protein-2 (MIP-2) and its receptor CXCR2.(2) Stimulated by the co-cultured irradiated lung epithelium, AMs produced a panel of inflammative molecules (IMs), such as cytokines (TNF-α, IL-6, IL-1α, IL-1β) and chemokines (MIP-2, MCP-1, LIX). TPL-treated AMs could reduce the production of these IMs. Meanwhile, AMs isolated from irradiated lung tissue secreted significantly high levels of IMs, which could be dramatically reduced by TPL.(3) TPL suppressed the phagocytosis of AMs as well as ROS production.Our results indicate that TPL mitigates radiation-induced pulmonary inflammation through the inhibition of the infiltration, IM secretion, and phagocytosis of AMs.

Topics: Antineoplastic Agents, Alkylating; Diterpenes; Epoxy Compounds; Humans; Inflammation; Macrophages, Alveolar; Phenanthrenes; Pneumonia

We investigated the synergic effects of components of particulate matter with aerodynamic diameters ≤2.5μm (PM2.5) on airway inflammation. Co-exposure to cadmium (Cd) and 9,10-phenanthrenequinone (9,10-PQ) additively/synergistically increased pro-inflammatory responses in airway epithelial cells, whereas co-exposure to Cd and phenanthrene resulted in no acceleration. These results suggest that the combination of metal and a quinone derivative can contribute to the exacerbation of respiratory diseases by PM2.5.

Topics: Air Pollutants; Cadmium; Cell Line; Drug Synergism; Epithelial Cells; Humans; Inflammation; Interleukin-6; Interleukin-8; Particulate Matter; Phenanthrenes; Reactive Oxygen Species

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

Chronic inflammation plays important roles in cancer initiation and progression. Resolving chronic inflammation or blocking inflammatory signal transduction may prevent cancer development. Here, we report that the combined low-dose use of two anti-inflammatory drugs, aspirin and triptolide, reduces spontaneous lung cancer incidence from 70% to 10% in a mouse model. Subsequent studies reveal that such treatment has little effect on resolving chronic inflammatory conditions in the lung, but it significantly blocks the NF-κB-mediated expression of proliferation and survival genes in cancer cells. Furthermore, triptolide and aspirin induce distinct mechanisms to potentiate each other to block NF-κB nuclear localization stimulated by inflammatory cytokines. While aspirin directly inhibits IκB kinases (IKKs) to phosphorylate IκBα for NF-κB activation, triptolide does not directly target IKKs or other factors that mediate IKK activation. Instead, it requires p53 to inhibit IκBα phosphorylation and degradation. Triptolide binds to and activates p38α and extracellular signal-regulated kinase 1/2 (ERK1/2), which phosphorylate and stabilize p53. Subsequently, p53 competes with IκBα for substrate binding to IKKβ and thereby blocks IκBα phosphorylation and NF-κB nuclear translocation. Inhibition of p38α and ERK1/2 or p53 mutations could abolish the inhibitory effects of triptolide on NF-κB. Our study defines a new p53-dependent mechanism for blocking NF-κB survival pathways in cancer cells.

Topics: Animals; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Aspirin; Cell Line, Tumor; Diterpenes; Epoxy Compounds; Female; Immunosuppressive Agents; Inflammation; Lung; Lung Neoplasms; Male; Mice; NF-kappa B; Phenanthrenes; Phosphorylation; Signal Transduction; Tumor Suppressor Protein p53

Surgeries, particularly ileocecal resection (ICR), are often required in the treatment of Crohn's disease (CD). However, recurrences are common for patients who undergo ICR, and anastomotic fibrosis is the main cause of re-operation. The present study aimed to investigate the therapeutic effects of triptolide (TPL) in ameliorating fibrosis following ileocolonic anastomosis. A model of IL‑10‑/‑ mice undergoing ICR was used to study postsurgical inflammation and fibrosis of anastomosis. For this purpsose, interleukin (IL)‑10‑/‑ mice were randomly divided into 3 groups as follows: the control group, the saline‑treated group subjected to ICR (ST‑ICR) and the TPL‑treated group subjected to ICR (TT‑ICR). Wild‑type (WT) mice of matching ages were assigned to the WT group. The effects of TPL treatment on ileocolonic anastomosis were determined by histopathological evaluation, western blot analysis and ELISA. The analysis of the effects of TPL treatment on microRNA‑16‑1 (miR‑16‑1) and heat shock protein 70 (HSP70) expression was carried out by RT‑qPCR and western blot analysis. Compared with the control group, significantly higher inflammation scores following anastomosis were observed in the ST‑ICR group (P<0.05), although reversion was observed in the TT‑ICR group, which was consistent with changes in the area of CD4+ cell infiltration. The elevated fibrosis scores and the overexpression of procollagen I and III in the ST‑ICR group were all inhibited by TPL. With an increase in the severity of inflammation and fibrosis, the levels of IL‑6, tumor necrosis factor‑α (TNF‑α) and transforming growth factor‑β1 (TGF‑β1) increased; however, a significant decrease in these levels was observed following treatment with TPL (P<0.05). The results of RT‑qPCR revealed that the upregulated miR‑16‑1 levels in the ST‑ICR group were significantly reduced by TPL. HSP70, which can be inhibited by miR-16-1, ameliorates anastomotic inflammation and fibrosis. Thus, the present study demonstrates that TPL exerts a protective effect against fibrosis following anastomosis in CD. The miR‑16‑1/HSP70 signaling pathway, which can be regulated by TPL, may thus represent a novel therapeutic option in CD that deserves further investigation.

Topics: Anastomosis, Surgical; Animals; Diterpenes; Epoxy Compounds; Gene Deletion; Gene Expression Regulation; HSP70 Heat-Shock Proteins; Inflammation; Interleukin-10; Intestines; Male; Mice; MicroRNAs; Phenanthrenes; Postoperative Complications; Protective Agents; Signal Transduction

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

Tanshinone IIA Sodium sulfonate (STS) is clinically used for treating cardiovascular diseases in Traditional Chinese Medicine due to its antioxidation and anti-inflammation activities. Intracellular chloride channel 1 (CLIC1) participates in the regulation of oxidative stress and inflammation. This study investigates whether CLIC1 mediates the cardioprotective effects of STS.. STS were used to treat atherosclerosis (AS) induced by feeding Apolipoprotein E-deficient (ApoE. STS treatment decreased atherosclerotic lesion area by 3.5 times (P = 0.001) in vivo. Meanwhile, STS reduced MDA production (13.6%, P = 0.008), increased SOD activity (113.6%, P = 0.008), decreased TNF-α (38.6%, P = 0.008) and IL-6 (43.0%, P = 0.03) levels, and downregulated the expression of CLIC1, ICAM-1, and VCAM-1 in the atherosclerotic mice. The dose-dependent anti-oxidative and anti-inflammatory effects of STS were further confirmed in vitro. Furthermore, CLIC1 depletion abolished the STS-mediated decrease of ROS and MDA production in HUVEC cells. Additionally, STS inhibited both CLIC1 membrane translocation and chloride ion concentration.. The anti-oxidant, and anti-inflammation properties of STS in preventing AS is mediated by its inhibition of CLIC1 expression and membrane translocation.

Topics: Animals; Antioxidants; Atherosclerosis; Cell Membrane; Chloride Channels; Down-Regulation; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Male; Mice; Oxidative Stress; Phenanthrenes; Protein Transport

Trauma resulted hemorrhagic shock (HS) leads to increased oxidative stress and inflammatory responses, which contributes greatly to organ failure or dysfunction. Tanshinone IIA sulfonate (TSA), as an antioxidant, may potentially be used in fluid resuscitation to prevent HS-induced organ damages.. In this study, a rat HS model was constructed. HS rats received TSA or vehicle drug during resuscitation. Mean arterial pressure and factors associated with organ failure or dysfunction, oxidative stress, and inflammatory response were investigated to evaluate treatment responses. Expression of proteins in NF-кB pathway was evaluated to elucidate the mechanism of TSA in preventing HS-induced organ damage.. Although HS induced organ damage and upregulated oxidative stress and inflammatory response, TSA treatment ameliorated organ dysfunction, reduced oxidative stress, and suppressed inflammatory responses. We also showed that TSA treatment attenuated HS-induced activation in NF-кB pathway.. TSA can potentially serve as an antioxidant for ameliorating HS-induced organ failure or function. Its mechanism of action may be through inhibiting NF-кB pathway.

Topics: Animals; Blood Pressure; Drug Evaluation, Preclinical; Drugs, Chinese Herbal; Inflammation; Male; Multiple Organ Failure; NF-kappa B; Oxidative Stress; Phenanthrenes; Phytotherapy; Rats, Wistar; Resuscitation; Shock, Hemorrhagic

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

Triptolide is a most important active ingredient extracted from traditional Chinese medicine Tripterygium, which has been widely used to treat glomerulonephritis as well as immune-mediated disorders, likely for its immunosuppressive, anti-proliferative and anti-inflammatory effects.. In this study, we have investigated the potential protective effects of triptolide against diabetic cardiomyopathy (DCM) by regulating immune system, attenuating inflammatory response, thus resulting in decreased cardiac fibrosis and improved left ventricle function.. Sprague-Dawley rats were randomly divided into 5 groups: normal group, diabetic group and diabetic rats treated with triptolide (50, 100, or 200μg/kg/day resp) for 8 weeks. Cardiac function was performed by echocardiography and histopathology of the hearts was examined with HE, Masson staining and scanning electron microscopy. Immune regulation mediator, macrophage infiltration, inflammatory response and cardiac fibrosis related cytokines were measured by RT-PCR, Western blot and Immunohistochemistry staining.. In the diabetic group, the expressions of TLR4 and NF-κB p65 were both up-regulated, which was associated with increased pro-inflammatory cytokines, coupled with cardiac fibrosis and impaired left ventricular function. Interestingly, pathological structure and function of left ventricle were both significantly improved in the triptolide treated groups. Furthermore, the immune mediator TLR4, downstream activator NF-κB p65, macrophage infiltration (CD68+), pro-inflammatory cytokines (TNF-α, IL-1β), cell adhesion molecule (VCAM-1) and chemokine (MCP-1) were significantly suppressed when treated with medium and high dosage triptolide compared with the diabetic group. Moreover, cardiac fibrosis pathway including α-SMA, TGF-β1, vimentin and collagen accumulations were observed significantly decreased in the triptolide treated groups.. Our data demonstrated that the protective effects of triptolide against DCM might attribute to inhibition of TLR4-induced NF-κB/IL-1β immune pathway, suppression of NF-κB/TNF-α/VCAM-1 inflammatory pathway and down-regulation of TGF-β1/α-SMA/Vimentin fibrosis pathway.

Topics: Animals; Autoimmune Diseases; Blood Glucose; Cardiomyopathies; Diabetic Cardiomyopathies; Diterpenes; Epoxy Compounds; Fibrosis; Heart Function Tests; Immunity, Innate; Inflammation; Male; Phenanthrenes; Rats; Rats, Sprague-Dawley; Toll-Like Receptor 4

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

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

Tripchlorolide (T4), an extract of the natural herb Tripterygium wilfordii Hook F, has been found to possess anti-inflammatory and immunosuppressive actions. In the current study, these actions were evaluated in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis by scoring the clinical signs, observing the infiltration of inflammatory cells and myelin sheath in the lumbar spinal cord of EAE mice. The results demonstrated that T4 (at a dose of 40 μg/kg) significantly reduced the severity of EAE and slowed down the ongoing EAE. Further analysis showed that T4 suppressed the mRNA and protein levels of the transcription factors T-bet and RoRrt and mRNA levels of IFN-γ and IL-17 in the spinal cords. Furthermore, T4 down-regulated the ERK1/2-NF-κB and JAK/STAT signaling pathways. At 40 μg/kg, T4 did not induce side effects on hematological parameters. These findings suggest that T4 ameliorates EAE by immunosuppression, providing a new insight into T4 application in multiple sclerosis treatment.

Topics: Animals; Demyelinating Diseases; Diterpenes; Down-Regulation; Encephalomyelitis, Autoimmune, Experimental; Female; Immunosuppressive Agents; Inflammation; Janus Kinases; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; NF-kappa B; Phenanthrenes; Signal Transduction; Spinal Cord; STAT1 Transcription Factor

Inflammatory pain is one of the most common clinical symptoms, mechanical allodynia and thermal hypersensitivities are associated with proinflammatory cytokines, and proinflammatory cytokine antagonists could alleviate the hypersensitivity. Previous studies showed that a traditional Chinese medicine ingredient, triptolide could inhibit inflammatory cytokines; however, it was still unknown whether triptolide had beneficial effects on treating inflammatory pain.. The effects of triptolide on Complete Freund's Adjuvant-induced acute inflammatory pain were investigated using behavioral tests. The activation of spinal glia was morphologically observed by immunofluorescent histochemistry. The levels of OX42, glia fibrillary acidic protein, and phosphorylated extracellular signal-regulated kinase in the spinal cord were detected by Western blot, and the messenger RNA levels of interleukin 1β, interleukin 6, and tumor necrosis factor alpha were detected by real-time polymerase chain reaction.. These results demonstrate that the triptolide effectively attenuates inflammatory pain induced by Complete Freund's Adjuvant, the underlying mechanism may regulate the phosphorylated extracellular signal-regulated kinase signaling pathway and inhibit the spinal glia activation, and then downregulate the proinflammatory cytokines; the triptolide may be clinically useful as a drug of anti-inflammatory pain.. In the present study, we first reported that repeated systemic administration of triptolide could safely prevent and reverse inflammatory pain. The triptolide may serve as a new potential compound for developing safe therapeutics for patients suffering inflammatory pain.

Topics: Acute Pain; Animals; Cytokines; Diterpenes; Drug Evaluation, Preclinical; Enzyme Activation; Epoxy Compounds; Extracellular Signal-Regulated MAP Kinases; Freund's Adjuvant; Immunosuppressive Agents; Inflammation; Inflammation Mediators; Male; Neuroglia; Phenanthrenes; Phytotherapy; Plant Extracts; Rats; Rats, Sprague-Dawley

Triptolide (TP) is a biologically active diterpene triepoxide from the Chinese herb Tripterygium wilfordii Hook f. Here, we identify and explore TAB1 as the binding target of TP in macrophages by using a comprehensive approach combining pull-down assays, in vitro assessments, and pharmaceutical and biological evaluation. We discover that TP inhibits TAK1 kinase activity by interfering with the formation of the TAK1-TAB1 complex, and the binding affinity of TP to TAB1 correlates highly with the inhibitory activity of TP against MAPK pathway activation in macrophages. We also find that the amino acid sequence between positions 373 and 502 of TAB1 is required for TP interaction. Our results suggest that TP could be a selective small-molecule inhibitor of the TAK1-TAB1 complex and that TAB1 could be a potential therapeutic target in inflammatory disease.

Topics: Adaptor Proteins, Signal Transducing; Animals; Cell Line; Cytokines; Diterpenes; Epoxy Compounds; Inflammation; Lipopolysaccharides; Macrophages; Mice; Phenanthrenes; Protein Binding; Recombinant Proteins; Tripterygium; Tumor Necrosis Factor-alpha

Cancer stem cells (CSCs) are affected by the local micro-environment, the niche, in which inflammatory stimuli and hypoxia act as steering factors. Here, two nuclear receptors (NRs) agonists, i.e. pioglitazone (PGZ), a ligand of peroxisome proliferator activated receptor-γ, and 6-OH-11-O-hydroxyphenanthrene (IIF), a ligand of retinoid X receptors, were investigated for their capability to interference with the cross-talk between breast CSCs and the niche compartment. We found that IIF potentiates the ability of PGZ to hamper the mammospheres-forming capability of human breast tumours and MCF7 cancer cells, reducing the expression of CSCs regulatory genes (Notch3, Jagged1, SLUG, Interleukin-6, Apolipoprotein E, Hypoxia inducible factor-1α and Carbonic anhydrase IX). Notably, these effects are not observed in normal-MS obtained from human breast tissue. Importantly, NRs agonists abolish the capability of hypoxic MCF7 derived exosomes to induce a pro-inflammatory phenotype in mammary glands fibroblasts. Moreover, NRs agonist also directly acts on breast tumour associated fibroblasts to downregulate nuclear factor-κB pathway and metalloproteinases (MMP2 and MMP9) expression and activity. In conclusion, NRs agonists disrupt the inflammatory cross-talk of the hypoxic breast CSCs niche.

Topics: Antigens, Neoplasm; Apolipoproteins E; Biomarkers, Tumor; Breast Neoplasms; Carbonic Anhydrase IX; Carbonic Anhydrases; Cell Hypoxia; Cell Survival; Exosomes; Female; Fibroblasts; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Inflammation; Ligands; Matrix Metalloproteinase 9; MCF-7 Cells; Models, Biological; Neoplastic Stem Cells; NF-kappa B; Phenanthrenes; Pioglitazone; PPAR gamma; Receptor Cross-Talk; Retinoid X Receptors; Spheroids, Cellular; Stem Cell Niche; Stromal Cells; Thiazolidinediones

Interleukin-1β (IL-1β) upregulates intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expressions in osteoarthritis fibroblast-like synoviocytes (OA-FLS) via nuclear factor (NF)-κB-mediated mechanism; enhancement of leukocyte infiltration and upregulation of proinflammatory mediators play a crucial role in OA pathophysiology. MicroRNA (miR)-146a suppresses inflammatory responses by inhibiting NF-κB activity and target gene expression, and epigenetic mechanisms are reportedly involved in miR expression regulation. Here, we aimed to verify the inhibition of ICAM-1/VCAM-1 expression in OA-FLS on denbinobin treatment and to determine whether this inhibition was due to the miR-146a-dependent pathway. We also assessed the epigenetic regulation caused by histone acetyltransferases involved in denbinobin action. Denbinobin attenuated the upregulation of IL-1β-induced ICAM-1/VCAM-1 expression and monocyte adhesion to OA-FLS. The mechanism underlying the inhibitory effects of denbinobin involved miR-146a induction, which in turn inhibited NF-κB signaling. This is because miR-146a inhibitor abrogated the inhibitory effects of denbinobin. Furthermore, histone acetyltransferase inhibitor attenuated the denbinobin-induced upregulation of miR-146a expression and inhibited the acetylation of NF-κB-binding sites located within the miR-146a promoter region. These data suggest that an epigenetic mechanism plays a crucial role in the upregulation of miR-146a expression in response to denbinobin treatment. Our overall findings suggest that denbinobin can be used as a potent anti-inflammatory agent.. Denbinobin inhibited IL-1β-induced ICAM-1/VCAM-1 expression and monocyte adhesion to OA-FLS. It was due to denbinobin increased miR-146a level, which in turn inhibited NF-κB signaling. Our overall findings suggest that denbinobin can be used as a potent anti-inflammatory agent.

Topics: Aged; Anthraquinones; Binding Sites; Cell Adhesion; Epigenesis, Genetic; Fibroblasts; Gene Expression Regulation; HeLa Cells; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-1beta; MicroRNAs; Microscopy, Fluorescence; Monocytes; NF-kappa B; Osteoarthritis; Patella; Phenanthrenes; Signal Transduction; Synovial Membrane; Synoviocytes; Vascular Cell Adhesion Molecule-1

Ischemia-reperfusion (I/R) injury after lung transplantation causes alveolar damage, lung edema, and acute rejection. Poly(adenosine diphosphate-ribose) polymerase (PARP) is a single-stranded DNA repair enzyme that induces apoptosis and necrosis after DNA damage caused by reactive oxygen species. We evaluated tissue protective effects of the PARP inhibitor (PARP-i) PJ34 against pulmonary I/R injury.. Rats (total n=45) underwent a thoracotomy with left hilar isolation and saline administration (sham group) or thoracotomy with hilar clamping and saline administration (I/R group) or PJ34 administration (PARP-i group). Parameters were measured for 7 days after reperfusion.. Pathologic analysis revealed that reperfusion injury was drastically suppressed in the PARP-i group 2 days after reperfusion. Terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick-end labeling-positive cells were significantly decreased in the PARP-i group compared to the I/R group (P<0.05). Accordingly, the wet-to-dry lung ratio in the I/R group was significantly higher compared with the PARP-i group (P=0.025). Four hours after reperfusion, serum tissue necrosis factor-α and interleukin-6 were significantly suppressed in the PARP-i group compared with the I/R group (P<0.05). Serum derivatives of reactive oxygen metabolites increased quickly and remained high in the I/R and PARP-i groups from 4 hr until 7 days after reperfusion. Interestingly, the serum biologic antioxidant potential in the PARP-i group was significantly higher than that in the I/R group from day 2 until day 7.. The PARP-i decreased inflammation and tissue damage caused by pulmonary I/R injury. These beneficial effects of the PARP-i may be correlated with its antioxidative efficacy.

Topics: Adenosine Triphosphate; Animals; Antioxidants; Apoptosis; DNA Repair; Inflammation; Interleukin-6; Lung; Lung Transplantation; Male; Oxidative Stress; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Rats, Wistar; Reactive Oxygen Species; Reperfusion Injury; Thoracotomy; Time Factors; Tumor Necrosis Factor-alpha

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

Cultivated Dendrobium denneanum has been substituted for other endangered Dendrobium species in recent years, but there have been few studies regarding either its chemical constituents or pharmacological effects. In this study, three phenanthrene glycosides, three 9,10-dihydrophenanthrenes, two 9,10-dihydrophenanthrenes glycosides, and four known phenanthrene derivatives, were isolated from the stems of D. denneanum. Their structures were elucidated on the basis of MS and NMR spectroscopic data. Ten compounds were found to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated mouse macrophage RAW264.7 cells with IC50 values of 0.7-41.5 μM, and exhibited no cytotoxicity in RAW264.7, HeLa, or HepG2 cells. Additionally, it was found that 2,5-dihydroxy-4-methoxy-phenanthrene 2-O-β-d-glucopyranoside, and 5-methoxy-2,4,7,9S-tetrahydroxy-9,10-dihydrophenanthrene suppressed LPS-induced expression of inducible NO synthase (iNOS) inhibited phosphorylation of p38, JNK as well as mitogen-activated protein kinase (MAPK), and inhibitory kappa B-α (IκBα). This indicated that both compounds exert anti-inflammatory effects by inhibiting MAPKs and nuclear factor κB (NF-κB) pathways.

Topics: Animals; Anti-Inflammatory Agents; Dendrobium; Glycosides; HeLa Cells; Hep G2 Cells; Humans; I-kappa B Proteins; Inflammation; Inhibitory Concentration 50; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; Macrophages; Mice; NF-kappa B; NF-KappaB Inhibitor alpha; Nitric Oxide; Nitric Oxide Synthase Type II; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Phosphorylation; Phytotherapy; Plant Extracts; Plant Stems

The objective of this study is to investigate the effect of local inflammation suppression on the bone regeneration. Gelatin hydrogels incorporating mixed immunosuppressive triptolide-micelles and bone morphogenic protein-2 (BMP-2) were prepared. The controlled release of both the triptolide and BMP-2 from the hydrogels was observed under in vitro and in vivo conditions. When either J774.1 macrophage-like or MC3T3-E1 osteoblastic cells were cultured in the hydrogels incorporating mixed 2.5, 5 or 10 mg of triptolide-micelles and BMP-2, the expression level of pro- and anti-inflammatory cytokines including interleukin (IL)-6 and IL-10 was down-regulated, but the alkaline phosphatase (ALP) activity was promoted compared with those of hydrogels incorporating BMP-2 without triptolide-micelles. When implanted into a critical-sized bone defect of rats, the hydrogels incorporating mixed 2.5 or 5 mg of triptolide-micelles and BMP-2 showed significantly lower number of neutrophils, lymphocytes, macrophages or dendritic and mast cells infiltrated into the defect, and lower expression level of IL-6, TNF-α, and IL-10 than those incorporating BMP-2 without triptolide-micelles. The reduced local inflammation responses at the defects implanted with the hydrogels incorporating mixed 2.5 or 5 mg of triptolide-micelles and BMP-2 subsequently enhanced the bone regeneration thereat. It is concluded that the proper local modulation of inflammation responses is a promising way to achieve the enhanced bone regeneration.

Topics: Animals; Bone Morphogenetic Protein 2; Bone Regeneration; Cell Line; Cytokines; Diterpenes; Epoxy Compounds; Gelatin; Hydrogels; Inflammation; Male; Mice; Micelles; Phenanthrenes; Rats; Rats, Wistar

Airway smooth muscle (ASM) plays an important immunomodulatory role in airway inflammation in asthma. In our previous in vitro studies in ASM cells delineating the pro-inflammatory mitogen-activated protein kinase (MAPK) signaling pathways activated by tumor necrosis factor α (TNFα), we observed that TNFα concomitantly induces the rapid, but transient, upregulation of the anti-inflammatory protein-mitogen-activated protein kinase phosphatase 1 (MKP-1). As this was suggestive of a negative feedback loop, the aim of this study was to investigate the molecular mechanisms of MKP-1 upregulation by TNFα and to determine whether MKP-1 is a negative feedback effector that represses MAPK-mediated pro-inflammatory signaling pathways and cytokine secretion in ASM cells. Herein, we show that TNFα increases MKP-1 mRNA expression and protein upregulation in a p38 MAPK-dependent manner. TNFα does not increase MKP-1 transcription (measured by MKP-1 promoter activity); rather, we found that TNFα-induced MKP-1 mRNA stability is regulated by the p38 MAPK pathway. Inhibiting MKP-1 upregulation (with triptolide) demonstrated the precise temporal control exerted on MAPK signaling by MKP-1. In the absence of MKP-1, downstream phosphoprotein targets of MAPKs (such as MSK-1 and histone H3) are not turned off at the right time, allowing pro-inflammatory pathways to continue in an unrestrained manner. This is confirmed by knocking-down MKP-1 by siRNA where enhanced secretion of the neutrophil chemoattractant cytokine-interleukin 8 was detected in the absence of MKP-1. Thus, by activating p38 MAP kinase, TNFα concomitantly upregulates the MAPK deactivator MKP-1 to serve as an important negative feedback effector, limiting the extent and duration of pro-inflammatory MAPK signaling and cytokine secretion in ASM cells.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Bronchi; Diterpenes; Dual Specificity Phosphatase 1; Epoxy Compounds; Feedback, Physiological; Gene Expression Regulation; Histones; Humans; Imidazoles; Inflammation; Interleukin-8; Myocytes, Smooth Muscle; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Primary Cell Culture; Promoter Regions, Genetic; Pyridines; Ribosomal Protein S6 Kinases, 90-kDa; RNA Stability; RNA, Small Interfering; Signal Transduction; Tumor Necrosis Factor-alpha

Recent literature highlights the importance of pro-inflammatory cytokines in the biology of breast cancer stem cells (CSCs), unraveling differences with respect to their normal counterparts. Expansion of mammospheres (MS) is a valuable tool for the in vitro study of normal and cancer mammary gland stem cells. Here, we expanded MSs from human breast cancer and normal mammary gland tissues, as well from tumorigenic (MCF7) and non-tumorigenic (MCF10) breast cell lines. We observed that agonists for the retinoid X receptor (6-OH-11-O-hydroxyphenanthrene), retinoic acid receptor (all-trans retinoic acid (RA)) and peroxisome proliferator-activated receptor (PPAR)-γ (pioglitazone (PGZ)), reduce the survival of MS generated from breast cancer tissues and MCF7 cells, but not from normal mammary gland or MCF10 cells. This phenomenon is paralleled by the hampering of pro-inflammatory Nuclear Factor-κB (NF-κB)/Interleukin-6 (IL6) axis that is hyperactive in breast cancer-derived MS. The hindrance of such pathway associates with the downregulation of MS regulatory genes (SLUG, Notch3, Jagged1) and with the upregulation of the differentiation markers estrogen receptor-α and keratin18. At variance, the PPARα agonist Wy14643 promotes MS formation, upregulating NF-κB/IL6 axis and MS regulatory genes. These data reveal that nuclear receptors agonists (6-OH-11-O-hydroxyphenanthrene, RA, PGZ) reduce the inflammation dependent survival of breast CSCs and that PPARα agonist Wy14643 exerts opposite effects on this phenotype.

Topics: Breast Neoplasms; Cell Line, Tumor; Cell Survival; Female; Humans; Inflammation; Interleukin-6; Neoplastic Stem Cells; NF-kappa B; Phenanthrenes; Pioglitazone; PPAR gamma; Pyrimidines; Receptors, Cytoplasmic and Nuclear; Receptors, Retinoic Acid; Retinoid X Receptors; Thiazolidinediones; Tretinoin

The commercial preparation named “Tripterygium glycosides” prepared by column chromatography has been used for the treatment of inflammatory and autoimmune diseases with significant efficacy but concurrent toxicity. The aim of this study was to reduce the toxicity of Tripterygium extracts, using cytotoxicity and anti-inflammatory activity of the three principal active components of Tripterygium wilfordii Hook. F. (TWHF)as guiding parameters. Column chromatography was replaced by sodium carbonate extraction for removing the acidic compounds and enriching epoxyditerpenoids and alkaloids in the extract. Results showed that the therapeutic index (IC50/EC50) on murine macrophage Raw 264.7 cells and rat mesangial HBZY-1 cells of the extract prepared by sodium carbonate extraction was significantly higher than that of Tripterygium glycosides(0.8 and 5.2 vs. 0.3 and 2.6, p < 0.05), while its cytotoxicity on human liver HL7702 cells was significantly lower (14.5 ± 1.4 vs. 6.8 ± 0.9, p < 0.05). Further acute oral toxicity experiments showed that the LD50 value of this extract was 1,210 mg/kg compared to 257 mg/kg for Tripterygium glycosides. All the above results suggest that Tripterygium extract prepared by sodium carbonate extraction may represent a potentially optimal source of medicine with good therapeutic index.

Topics: Animals; Anti-Inflammatory Agents; Autoimmune Diseases; Carbonates; Cell Line; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Female; Humans; Inflammation; Lactones; Male; Mice; Pentacyclic Triterpenes; Phenanthrenes; Phytotherapy; Pyridines; Rats; Tripterygium; Triterpenes

Cryptotanshinone (CTS), a major constituent extracted from the medicinal herb Salvia miltiorrhiza Bunge, has well-documented antioxidative and anti-inflammatory effects. In the present study, the pharmacological effects and underlying molecular mechanisms of CTS on lipopolysaccharide (LPS)-induced inflammatory responses were investigated. By enzyme-linked immunosorbent assay, we observed that CTS reduced significantly the production of proinflammatory mediators (tumor necrosis factor-α and interleukin-6) induced by LPS in murine macrophage-like RAW264.7 cells. Mechanistically, CTS inhibited markedly the phosphorylation of mitogen-activated protein kinases (MAPKs), including ERK1/2, p38MAPK, and JNK, which are crucially involved in regulation of proinflammatory mediator secretion. Moreover, immunofluorescence and western blot analysis indicated that CTS abolished completely LPS-triggered nuclear factor-κB (NF-κB) activation. Taken together, these data implied that NF-κB and MAPKs might be the potential molecular targets for clarifying the protective effects of CTS on LPS-induced inflammatory cytokine production in macrophages.

Topics: Animals; Anti-Inflammatory Agents; Blotting, Western; Cell Line; Enzyme-Linked Immunosorbent Assay; Fluorescent Antibody Technique; Inflammation; Interleukin-6; Lipopolysaccharides; Macrophages; MAP Kinase Signaling System; Mice; Mitogen-Activated Protein Kinases; NF-kappa B; Phenanthrenes; Phosphorylation; Signal Transduction; Tumor Necrosis Factor-alpha

Inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) have been suggested to play important roles in various inflammatory diseases. We explored the anti-inflammatory potential of a natural compound, denbinobin (5-hydroxy-3,7-dimethoxy-1,4-phenanthraquinone), by examining its effects on the expression and activity of iNOS and COX-2 in LPS-activated macrophages. Denbinobin markedly decreased the LPS (1 μg/mL)-induced increase in iNOS and COX-2 gene and protein expression, as well as levels of the downstream products NO and prostaglandin E2, in a concentration-dependent manner (0.3-3 μM). In clarifying the mechanisms involved, denbinobin was found not only to inhibit LPS-induced nuclear factor κB (NF-κB) activation, an effect highly correlated with its inhibitory effect on LPS-induced inhibitory κB kinase activation, inhibitory κB degradation, NF-κB phosphorylation, and binding of NF-κB to the κB motif of the iNOS and COX-2 promoters, but also suppressed phosphorylation of mitogen-activated protein kinases. Reporter gene assays and Western blotting revealed that denbinobin significantly suppressed NF-κB activation. Furthermore, denbinobin also downregulated the LPS-mediated CD14/toll-like receptor 4 complex level and TNF-α, IL-1β, and IL-10 mRNA expression. Our results demonstrate that denbinobin exerts potent anti-inflammatory activity, suggesting that it might provide a new therapeutic approach to inflammatory diseases.

Topics: Animals; Anthraquinones; Anti-Inflammatory Agents; Cell Line; Cyclooxygenase 2; Cytokines; Dinoprostone; Gene Expression Regulation, Enzymologic; Inflammation; Lipopolysaccharides; Macrophages; Mice; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Phenanthrenes; Phosphorylation; Response Elements

Triptolide, a diterpene triepoxide, from the Chinese herb Tripterygium wilfordii Hook.f, exerts its anti-inflammatory and immunosuppressive activities by inhibiting the transcription factor nuclear factor-κB (NF-κB) pathway, through a mechanism not yet fully understood. We found that triptolide, in nanomolar concentrations, suppressed both constitutive and inducible NF-κB activation, but did not directly inhibit binding of p65 to the DNA. The diterpene did block TNF-induced ubiquitination, phosphorylation, and degradation of IκBα, the inhibitor of NF-κB and inhibited acetylation of p65 through suppression of binding of p65 to CBP/p300. Triptolide also inhibited the IκBα kinase (IKK) that activates NF-κB and phosphorylation of p65 at serine 276, 536. Furthermore, the NF-κB reporter activity induced by TNF-TNFR1-TRADD-TRAF2-NIK-TAK1-IKKβ was abolished by the triepoxide. Triptolide also abrogated TNF-induced expression of cell survival proteins (XIAP, Bcl-x(L), Bcl-2, survivin, cIAP-1 and cIAP-2), cell proliferative proteins (cyclin D1, c-myc and cyclooxygenase-2), and metastasis proteins (ICAM-1 and MMP-9). This led to enhancement of apoptosis induced by TNF, taxol, and thalidomide by the diterpene and to suppression of tumor invasion. Overall, our results demonstrate that triptolide can block the inflammatory pathway activated by TNF-TNFR1-TRADD-TRAF2-NIK-TAK1-IKK, sensitizes cells to apoptosis, and inhibits invasion of tumor cells.

Topics: Antineoplastic Agents; Cell Line; Cell Proliferation; Diterpenes; Enzyme Inhibitors; Epoxy Compounds; Gene Expression Regulation, Neoplastic; Histone Acetyltransferases; Humans; Inflammation; Leukemia; Molecular Structure; Neoplasm Invasiveness; Neoplasm Metastasis; NF-kappa B; Paclitaxel; Phenanthrenes; Signal Transduction; Thalidomide; Tumor Necrosis Factor-alpha

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

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

Dietary flavonoids have various biological functions, and there is increasing evidence that reduced prevalence and severity of allergic reactions are associated with the intake of flavonoids. Among natural flavonoids, apigenin is a potent anti-inflammatory agent. However, the mechanisms of apigenin's effect remain uncertain. Monocyte-derived chemokine (MDC) plays a pivotal role in recruiting T-helper (Th) 2 cells in the allergic inflammation process. In the late phase of allergic inflammation, the Th1 chemokine interferon-inducible protein 10 (IP-10) has also been found in elevated levels in the bronchial alveolar fluid of asthmatic children. We used human THP-1 monocyte cells, pretreated with or without apigenin, prior to lipopolysaccharide stimulation. By means of enzyme-linked immunosorbent assay, we found that apigenin inhibited production of both MDC and IP-10 by THP-1 cells and that the suppressive effect of apigenin was not reversed by the estrogen receptor antagonist ICI182780. The p65 phosphorylation of nuclear factor kappaB remained unaffected, but the phosphorylation of p38, c-Jun N-terminal kinase, and extracellular signal-regulated kinase mitogen-activated protein kinase pathways were all blocked. We found that inhibition of c-raf phosphorylation might be the target of apigenin's anti-inflammation property.

Topics: Anti-Inflammatory Agents; Apigenin; Bronchioles; Bronchoalveolar Lavage Fluid; Chemokine CXCL10; Chemokines; eIF-2 Kinase; Enzyme-Linked Immunosorbent Assay; Estrogen Antagonists; Humans; Hypersensitivity; Inflammation; Lipopolysaccharides; Mitogen-Activated Protein Kinases; Monocytes; NF-kappa B; Phenanthrenes; Phosphorylation; Plant Extracts; Proto-Oncogene Proteins c-raf; Receptors, Estrogen; Th1 Cells; Th2 Cells

9,10-Dihydro-2,5-dimethoxyphenanthrene-1,7-diol (RSCL-0520) is a phenanthrene isolated from Eulophia ochreata, one of the Orchidaceae family, known by local tradition to exhibit medicinal properties. However, no anti-inflammatory activity or any molecular mechanisms involved have been reported or elucidated. Here, for the first time, we evaluate the anti-inflammatory properties of RSCL-0520 on responses induced by lipopolysaccharide (LPS) and mediated via Toll-like receptors (TLRs).. The in vitro anti-inflammatory activities of RSCL-0520 were investigated in LPS-stimulated monocytic cells, measuring activation of cytokine and inflammatory genes regulated by nuclear factor-kappaB (NF-kappaB). Tumour necrosis factor (TNF)-alpha levels in serum following LPS stimulation in mice and carrageenan-induced paw oedema in rats were used as in vivo models.. Pretreatment with RSCL-0520 effectively inhibited LPS-induced, TLR4-mediated, NF-kappaB-activated inflammatory genes in vitro, and reduced both LPS-induced TNF-alpha release and carrageenan-induced paw oedema in rats. Treatment with RSCL-0520 reduced LPS-stimulated mRNA expression of TNF-alpha, COX-2, intercellular adhesion molecule-1, interleukin (IL)-8 and IL-1beta, all regulated through NF-kappaB activation. RSCL-0520, however, did not interfere with any cellular processes in the absence of LPS.. RSCL-0520 blocked signals generated by TLR4 activation, as shown by down-regulation of NF-kappaB-regulated inflammatory cytokines. The inhibitory effect involved both MyD88-dependent and -independent signalling cascades. Our data elucidated the molecular mechanisms involved, and support the search for plant-derived TLR antagonists, as potential anti inflammatory agents.

Topics: Animals; Cell Line; Dose-Response Relationship, Drug; Down-Regulation; Edema; Female; Humans; Inflammation; Inflammation Mediators; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Monocytes; Orchidaceae; Phenanthrenes; Rats; Rats, Wistar; Signal Transduction; Toll-Like Receptors; Tumor Necrosis Factor-alpha

The brain inflammatory response induced by stroke contributes to cell death and impairs neurogenesis. Poly(ADP-ribose) polymerase-1 (PARP-1) is a coactivator of the transcription factor NF-kappaB and required for NF-kappaB-mediated inflammatory responses. Here we evaluated PARP inhibition as a means of suppressing post-stroke inflammation and improving outcome after stroke. Rats were subjected to bilateral carotid occlusion-reperfusion, and treatment with the PARP inhibitor N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide (PJ34) was begun 48 h later. PJ34 was found to rapidly suppress the ischemia-induced microglial activation and astrogliosis. Behavioral tests performed 6 to 8 weeks after ischemia showed deficits in spatial memory and learning that were lessened by the PJ34 treatment. Immunohistochemical evaluation of hippocampus at 8 weeks after ischemia showed increased neuronal density in CA1 layer of PJ34-treated animals relative to vehicle-treated animals. Bromodeoxyuridine labeling showed formation of new neurons in hippocampal CA1 area in PJ34-treated animals, but not in vehicle-treated animals. Together, these results suggest that treatment with a PARP inhibitor for several days after ischemia enhances long-term neuronal survival and neurogenesis by reducing inflammation.

Topics: Animals; Brain Ischemia; Cell Survival; Enzyme Inhibitors; Hippocampus; Inflammation; Learning; Memory; Neurons; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Stroke; Treatment Outcome

The aim of this study was to explore the inhibitory effects of Tanshinone II(A) on the production of proinflammation cytokines in radiation-stimulated microglia.. Microglia cells were treated with 2, 4, 8, 16, and 32 Gy of irradiation or sham-irradiated in the presence or absence of 1.0 microg/mL of Tanshinone II(A). The effects of Tanshinone II(A) on radiation-induced proinflammatory cytokines were evaluated by real-time polymerase chain reaction; the expression level of nuclear factor (NF-kappabeta) p65 in cytoplasm and nucleus was measured by Western blot. Immunofluorescence staining and confocal microscopy analysis were applied to detect the expression of gamma-H2AX and p65 postirradiation.. Radiation-induced release of proinflammatory cytokines in BV-2 cells was detectable after irradiation. Tanshinone II(A) decreased the radiation-induced release of proinflammatory cytokines. Further, Western blotting showed that Tanshinone II(A) could attenuate the nuclear translocation of (NF-kappabeta) p65 submit postirradiation. Immunofluorescence staining showed gamma-H2AX foci formation with p65 translocation into the nucleus postirradiation.. Our data indicated that Tanshinone II(A) exerts anti-inflammatory properties by suppressing the transcription of proinflammatory cytokine genes that might be associated with the NF-kappabeta signaling pathway. It is postulated that irradiation causes immediate cellular reaction, and that double-strand breaks trigger the molecular response that leads to NF-kappabeta pathway activation.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antibodies, Monoclonal; Cell Survival; Cells, Cultured; DNA Primers; Inflammation; Mice; Microglia; Phenanthrenes; Polymerase Chain Reaction; Radionuclide Imaging; Radiotherapy; RNA

Activated microglia participate in neuroinflammation which contributes to neuronal damage in neurodegenerative diseases. Inhibition of microglial activation may have potential anti-inflammatory effects. Our laboratory has previously reported that triptolide, a natural biologically active compound extracted from Tripterygium wilfordii, could protect dopaminergic neurons from inflammation-mediated damage. However, the mechanism by which triptolide inhibits inflammation remains unknown. We reported here that inhibition of prostaglandin E(2) (PGE(2)) production could be a potential mechanism of triptolide to suppress inflammation. Triptolide suppressed c-jun NH2-terminal kinase (JNK) phosphorylation, cyclooxygenase 2 (COX-2) expression and PGE(2) production in microglial cultures treated with lipopolysaccharide (LPS). Triptolide also greatly inhibited the transcriptional activity, but not the DNA-binding activity of nuclear factor-kappaB (NF-kappaB) in microglia following LPS stimulation. These results indicate that triptolide might suppress NF-kappaB activity to down-regulate COX-2 expression. The LPS-stimulated transcriptional activity of NF-kappaB was suppressed by inhibition of p38MAPK, but not by that of JNK and extracellular signal-regulated kinase. Furthermore, the LPS-induced PGE(2) production was reduced by inhibiting these kinases. Taken together, these results suggest that triptolide may suppress neuroinflammation via a mechanism that involves inactivation of two parallel signaling pathways: p38-NF-kappaB-COX-2-PGE(2) and JNK-PGE(2).

Topics: Animals; Blotting, Western; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Diterpenes; Electrophoretic Mobility Shift Assay; Epoxy Compounds; Immunosuppressive Agents; Inflammation; Lipopolysaccharides; MAP Kinase Kinase 4; Microglia; NF-kappa B; Phenanthrenes; Rats; Rats, Sprague-Dawley; Signal Transduction; Transfection

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

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

Triptolide (TPT), a diterpenoid triepoxide, is the major component isolated from the Chinese herb Tripterygium wilfordii Hook. f. Previous studies have shown that TPT has immunosuppressive properties and is effective in prolonging graft survival and suppressing autoimmune responses. The aim of this study was to investigate the protective effect of TPT in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. Treatment of C57BL/6 mice with TPT from the date of EAE induction significantly delayed EAE onset and suppressed disease severity, accompanied with reduced inflammation and demyelination in the central nervous system. TPT treatment lead to a significant inhibition of the mRNA expression of both Th1/Th(IL-17) and Th2 cytokines in spleen mononuclear cells (MNC) as well as in spinal cord tissues. In addition, the expression of Forkhead box p3 (Foxp3) was up-regulated in spleen MNC after TPT treatment. Furthermore, we detected apparent inhibition of nuclear factor-kappa B (NF-kappaB)-DNA binding activity, increased expression of the inhibitor of nuclear factor-kappa Balpha (IkappaBalpha) and decreased expression of pIkappaBalpha in spleen MNC in TPT-treated EAE mice. Taken together, these findings indicate that TPT has profound immunoregulatory functions and potential protective values for the treatment of autoimmune inflammatory disorders.

Topics: Animals; Blotting, Western; Cell Proliferation; Cytokines; Demyelinating Diseases; Diterpenes; Electrophoretic Mobility Shift Assay; Encephalomyelitis, Autoimmune, Experimental; Epoxy Compounds; Forkhead Transcription Factors; Gene Expression; Immunosuppressive Agents; Inflammation; Leukocytes, Mononuclear; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Phenanthrenes; Reverse Transcriptase Polymerase Chain Reaction; Spinal Cord; T-Lymphocytes

Chinese herbal remedy Tripterygium wilfordii Hook. f. (TWHF) has been reported to be therapeutically efficacious in the treatment of rheumatoid arthritis (RA), but its in vivo actions have not been clarified. The purpose of this study was to investigate the effects of triptolide, a diterpenoid triepoxide extracted from TWHF, on inflammation and cartilage destruction in collagen-induced arthritis (CIA) model mice. Histological examination demonstrated that triptolide significantly reduced the inflammatory responses and cartilage damage in the joint tissues. Interestingly, triptolide interfered with CIA-augmented expression of matrix metalloproteinases-13 and -3, which are considered to be key enzymes in the pathological destruction of cartilage, and simultaneously augmented CIA-reduced tissue inhibitors of metalloproteinases-1 and -2 expression in the joints. Moreover, triptolide inhibited prostaglandin E(2) production via selective suppression of the production and gene expression of cyclooxygenase (COX)-2, but not COX-1. The levels of interleukin (IL)-1beta, tumor necrosis factor alpha and IL-6 were also decreased by triptolide in the joint tissues and sera as well as the suppression of CIA-mediated expression of their mRNAs in the joints. In addition, triptolide treatment in vivo was able to reduce an abundance of nuclear factor-kappaB, the transcriptional factor closely related to the inflammatory process, in articular cartilage and synovium in CIA mice. These results suggest that triptolide exerts novel chondroprotective and anti-inflammatory effects on RA, and the therapeutic action of TWHF on RA is, in part, due to the triptolide activities.

Topics: Animals; Arthritis, Experimental; Cartilage; Collagen; Cytokines; Dinoprostone; Disease Progression; Diterpenes; Epoxy Compounds; Gene Expression; Immunohistochemistry; In Situ Hybridization; Inflammation; Male; Mice; Mice, Inbred DBA; NF-kappa B; Phenanthrenes; Reverse Transcriptase Polymerase Chain Reaction

Most stroke patients do not present for medical treatment until several hours after onset of brain ischemia. Consequently, neuroprotective strategies are required with comparably long therapeutic windows. Poly(ADP-ribose) polymerase inhibitors such as PJ34 are known to suppress microglial activation, a postischemic event that may contribute to neuronal death. We evaluated the effects of PJ34 administered 8 hours after transient forebrain ischemia.. Rats were subjected to 10 minutes of forebrain ischemia and treated with PJ34 for 7 days beginning 8 hours after reperfusion. Activated microglia and infiltrating macrophages were evaluated at serial time points between zero and 14 days after ischemia by immunostaining for CD11b. CA1 neuronal survival was evaluated 7 days after ischemia.. Rats treated with PJ34 showed a near-complete inhibition of microglia/macrophage activation (evaluated on day 5) and an 84% reduction in CA1 neuronal death.. Administration of PJ34 as late as 8 hours after transient ischemia-reperfusion has a large protective effect on CA1 survival. This effect may be mediated by suppression of the postischemic brain inflammatory response.

Topics: Animals; Brain Ischemia; Cell Death; Inflammation; Male; Neurons; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Rats; Rats, Sprague-Dawley; Reperfusion

A hallmark of rheumatoid- and osteoarthritis (OA) is proinflammatory cytokine-induced degeneration of cartilage collagen and aggrecan by matrix metalloproteinases (MMPs) and aggrecanases (ADAMTS). Effects of the Chinese herb, Tripterygium wilfordii Hook F (TWHF), on cartilage and its anti-arthritic mechanisms are poorly understood. This study investigated the impact of a purified derivative of TWHF, PG490 (triptolide), on cytokine-stimulated expression of the major cartilage damaging proteases, MMP-3, MMP-13, and ADAMTS4. PG490 inhibited cytokine-induced MMP-3, MMP-13 gene expression in primary human OA chondrocytes, bovine chondrocytes, SW1353 cells, and human synovial fibroblasts. Triptolide was effective at low doses and blocked the induction of MMP-13 by IL-1 in human and bovine cartilage explants. TWHF extract and PG490 also suppressed IL-1-, IL-17-, and TNF-alpha-induced expression of ADAMTS-4 in bovine chondrocytes. Thus, PG490 could protect cartilage from MMP- and aggrecanase-driven breakdown. The immunosuppressive, cartilage protective, and anti-inflammatory properties could make PG490 potentially a new therapeutic agent for arthritis.

Topics: ADAM Proteins; ADAMTS4 Protein; Animals; Cartilage; Cattle; Cells, Cultured; Chondrocytes; Collagenases; Cytokines; Diterpenes; Dose-Response Relationship, Drug; Down-Regulation; Epoxy Compounds; Gene Expression Regulation; Humans; Inflammation; Interleukin-1; Interleukin-17; Matrix Metalloproteinase 13; Matrix Metalloproteinase 3; Matrix Metalloproteinase Inhibitors; Metalloendopeptidases; Phenanthrenes; Plant Extracts; Procollagen N-Endopeptidase; RNA, Messenger; Tripterygium; Tumor Necrosis Factor-alpha

Mounting lines of evidence have suggested that brain inflammation participates in the pathogenesis of Parkinson's disease. Triptolide is one of the major active components of Chinese herb Tripterygium wilfordii Hook F, which possesses potent anti-inflammatory and immunosuppressive properties. We found that triptolide concentration-dependently attenuated the lipopolysaccharide (LPS)-induced decrease in [3H]dopamine uptake and loss of tyrosine hydroxylase-immunoreactive neurons in primary mesencephalic neuron/glia mixed culture. Triptolide also blocked LPS-induced activation of microglia and excessive production of TNFalpha and NO. Our data suggests that triptolide may protect dopaminergic neurons from LPS-induced injury and its efficiency in inhibiting microglia activation may underlie the mechanism.

Topics: Analysis of Variance; Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Count; Cells, Cultured; Diterpenes; Dopamine; Dose-Response Relationship, Drug; Drug Interactions; Drugs, Chinese Herbal; Embryo, Mammalian; Enzyme-Linked Immunosorbent Assay; Epoxy Compounds; Female; Immunohistochemistry; Inflammation; Lipopolysaccharides; Male; Mesencephalon; Microglia; Neurons; Nitrites; Phenanthrenes; Pregnancy; Rats; Tritium; Tumor Necrosis Factor-alpha; Tyrosine 3-Monooxygenase

Mitogen-activated protein (MAP) kinases play a pivotal role in the macrophages in the production of proinflammatory cytokines triggered by lipopolysaccharides. However, their function in the responses of macrophages to Gram-positive bacteria is poorly understood. Even less is known about the attenuation of MAP kinase signaling in macrophages exposed to Gram-positive bacteria. In the present study, we have investigated the regulation of MAP kinases and the role of MAP kinase phosphatase (MKP)-1 in the production of pro-inflammatory cytokines using murine RAW264.7 and primary peritoneal macrophages after peptidoglycan stimulation. Treatment of macrophages with peptidoglycan resulted in a transient activation of JNK, p38, and extracellular signal-regulated kinase. Most interestingly, MKP-1 expression was potently induced by peptidoglycan, and this induction was concurrent with MAP kinase dephosphorylation. Triptolide, a diterpenoid triepoxide, potently blocked the induction of MKP-1 by peptidoglycan and prolonged the activation of JNK and p38. Overexpression of MKP-1 substantially attenuated the production of tumor necrosis factor (TNF)-alpha induced by peptidoglycan, whereas knockdown of MKP-1 by small interfering RNA substantially increased the production of both TNF-alpha and interleukin-1 beta. Finally, we found that in primary murine peritoneal macrophages, MKP-1 induction following peptidoglycan stimulation also coincided with inactivation of JNK and p38. Blockade of MKP-1 induction resulted in a sustained activation of both JNK and p38 in primary macrophages. Our results reveal that MKP-1 critically regulates the expression of TNF-alpha and interleukin-1 beta in RAW264.7 cells and further suggest a central role for this phosphatase in controlling the inflammatory responses of primary macrophages to Gram-positive bacterial infection.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Cycle Proteins; Cell Line; Diterpenes; Dual Specificity Phosphatase 1; Enzyme Activation; Epoxy Compounds; Extracellular Signal-Regulated MAP Kinases; Female; Gene Expression; Immediate-Early Proteins; Inflammation; Interleukin-1; JNK Mitogen-Activated Protein Kinases; Macrophages; Macrophages, Peritoneal; MAP Kinase Kinase 4; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase Kinases; p38 Mitogen-Activated Protein Kinases; Peptidoglycan; Phenanthrenes; Phosphoprotein Phosphatases; Phosphorylation; Protein Phosphatase 1; Protein Tyrosine Phosphatases; Transfection; Tumor Necrosis Factor-alpha

The lack of efficacy of anti-inflammatory drugs, anti-coagulants, anti-oxidants, etc. in critically ill patients has shifted interest towards developing alternative treatments. Since inhibitors of the nuclear enzyme poly-(ADP-ribose) polymerase (PARP) were found to be beneficial in many pathophysiological conditions associated with oxidative stress and PARP-1 knock-out mice proved to be resistant to bacterial lipopolysaccharide (LPS)-induced septic shock, PARP inhibitors are candidates for such a role. In this study, the mechanism of the protective effect of a potent PARP-1 inhibitor, PJ34 was studied in LPS-induced (20mg/kg, i.p.) septic shock in mice. We demonstrated a significant inflammatory response by magnetic resonance imaging in the dorsal subcutaneous region, in the abdominal regions around the kidneys and in the inter-intestinal cavities. We have found necrotic and apoptotic histological changes as well as obstructed blood vessels in the liver and small intestine. Additionally, we have detected elevated tumor necrosis factor-alpha levels in the serum and nuclear factor kappa B activation in liver of LPS-treated mice. Pre-treating the animals with PJ34 (10mg/kg, i.p.), before the LPS challenge, besides rescuing the animals from LPS-induced death, attenuated all these changes presumably by activating the phosphatidylinositol 3-kinase-Akt/protein kinase B cytoprotective pathway.

Topics: Animals; Base Sequence; Binding Sites; Consensus Sequence; Endotoxins; Enzyme Inhibitors; Escherichia coli; Inflammation; Lipopolysaccharides; Mice; Mice, Inbred BALB C; NF-kappa B; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Sepsis; Shock, Septic

Abietic acid is one of the terpenoids, which are multifunctional natural compounds. It has been reported that abietic acid suppresses effects on inflammation. However, the mechanism underlying the anti-inflammatory effects remains unclear. The present work indicates that abietic acid suppresses the protein expression of tumor necrosis factor-alpha and cyclooxygenase 2, which are involved in inflammation, in lipopolysaccharide-stimulated macrophages. Moreover, this effect resembles that of thiazolidinedione, a synthetic peroxisome proliferator-activated receptor-gamma (PPARgamma) ligand. Indeed, abietic acid activates PPARgamma in luciferase reporter assays. The activity of abietic acid induces PPARgamma target gene expression in RAW264.7 macrophages and 3T3-L1 adipocytes. These data indicate that abietic acid is a PPARgamma ligand and that its anti-inflammatory effect is partly due to the activation of PPARgamma in stimulated macrophages. The present work suggests a novel possibility that abietic acid, a naturally occurring compound, can be used not only for anti-inflammation but also for regulating lipid metabolism and atherosclerosis.

Topics: 3T3 Cells; Abietanes; Adipocytes; Animals; CD36 Antigens; Cells, Cultured; CREB-Binding Protein; Cyclooxygenase 2; Diterpenes; Gene Expression Regulation; Inflammation; Isoenzymes; Ligands; Lipid Metabolism; Lipopolysaccharides; Luciferases; Macrophages, Peritoneal; Male; Mice; Mice, Inbred BALB C; Nuclear Proteins; Phenanthrenes; Prostaglandin-Endoperoxide Synthases; Receptors, Cytoplasmic and Nuclear; Thiazoles; Thiazolidinediones; Trans-Activators; Transcription Factors; Tumor Necrosis Factor-alpha

In this study we evaluate the antifibrotic properties of PG-490-88, a water-soluble derivative of triptolide. Triptolide is an oxygenated diterpene that is derived from a traditional Chinese herb that has potent immunosuppressive and antitumor activity. We used the intratracheal bleomycin mouse model and found that PG490-88 inhibits fibrosis in the bleomycin group when given the same day or 5 days after bleomycin. PG490-88 also markedly reduced the number of myofibroblasts in the bleomycin treatment group. An enzyme-linked immunosorbent assay of transforming growth factor (TGF)-beta in the bronchoalveolar lavage fluid showed a significant decrease in TGF-beta in the PG490-88-treated groups compared to the bleomycin-treated group. Additionally, triptolide blocked bleomycin-induced increase in TGF-beta mRNA in cultured normal human lung fibroblasts. The efficacy of PG490-88 when administered late after bleomycin installation suggests a potential role in the treatment of idiopathic pulmonary fibrosis.

Topics: Animals; Bleomycin; Bronchoalveolar Lavage Fluid; Cell Survival; Diterpenes; Drugs, Chinese Herbal; Epoxy Compounds; Fibroblasts; Immunosuppressive Agents; Inflammation; Lung; Male; Mice; Mice, Inbred C57BL; Phenanthrenes; Pulmonary Fibrosis; Transcription, Genetic; Transforming Growth Factor beta

The anti-inflammatory activity of abietic acid, a diterpene isolated from Pimenta racemosa var. grissea (Myrtaceae), was evaluated in-vivo and in-vitro. This compound significantly inhibited rat paw oedema induced by carrageenan in a time- and dose-dependent manner, and mouse ear oedema induced by 12-O-tetradecanoylphorbol acetate, after oral or topical administration. The inhibition of myeloperoxidase enzyme showed that its topical activity was influenced by neutrophil infiltration into the inflamed tissues (ears). In addition, the effect of abietic acid on some macrophage functions was analysed in-vitro. Non-toxic concentrations of abietic acid inhibited prostaglandin E2 (PGE2) production in lipopolysaccharide-treated macrophages, whereas nitrite, tumour necrosis factor alpha and interleukin-1beta production were only weakly affected by this diterpene. PGE2 production from A23187-stimulated macrophages was only inhibited at high doses (100 microM) and it failed to modify leukotriene C4 production. These results indicate that abietic acid exerts in-vivo anti-inflammatory activity after oral or topical administration and has partial ability to prevent the production of some inflammatory mediators.

Topics: Abietanes; Administration, Oral; Administration, Topical; Animals; Cytokines; Diterpenes; Edema; Fibrinolytic Agents; Inflammation; Macrophages; Male; Mice; Neutrophil Infiltration; Phenanthrenes; Plant Extracts; Rats; Rats, Wistar

Monocyte chemoattractant protein 1 (MCP-1) has an important influence on monocyte migration into sites of inflammation. Our understanding of the signal transduction pathways involved in the response of monocytes to MCP-1 is quite limited yet potentially significant for understanding and manipulating the inflammatory response. Prior studies have demonstrated a crucial regulatory role for cytosolic phospholipase A(2) (cPLA(2)) in monocyte chemotaxis to MCP-1. In these studies we investigated the role for another PLA(2), calcium-independent PLA(2) (iPLA(2)) in comparison to cPLA(2). Pharmacological inhibitors of PLA(2) were found to substantially inhibit chemotaxis. Using antisense oligodeoxyribonucleotide treatment we found that iPLA(2) expression is required for monocyte migration to MCP-1. Complete blocking of the chemotactic response was observed with inhibition of either iPLA(2) or cPLA(2) expression by their respective antisense oligodeoxyribonucleotide. In reconstitution experiments, lysophosphatidic acid completely restored MCP-1-stimulated migration in iPLA(2)-deficient monocytes, whereas lysophosphatidic acid was without effect in restoring migration in cPLA(2)-deficient monocytes. To the contrary, arachidonic acid fully restored migration of cPLA(2)-deficient monocytes while having no effect on the iPLA(2)-deficient monocytes. Additional studies revealed that neither enzyme appears to be upstream of the other indicating that iPLA(2) and cPLA(2) represent parallel regulatory pathways. These data demonstrate novel and distinct roles for these two phospholipases in this critical step in inflammation.

Topics: Aminobenzoates; Arachidonic Acid; Arachidonic Acids; Aristolochic Acids; Chemokine CCL2; Chemotaxis, Leukocyte; Chlorobenzoates; Cinnamates; Enzyme Inhibitors; Fatty Acids; Group IV Phospholipases A2; Group VI Phospholipases A2; Humans; Inflammation; Lysophospholipids; Monocytes; Naphthalenes; Oligodeoxyribonucleotides, Antisense; ortho-Aminobenzoates; Phenanthrenes; Phospholipases A; Pyrones; Signal Transduction

We recently described a stimulatory effect of high doses (> 100 mumol/L) diclofenac on platelet adhesion. In this study we extend our research to the possible biochemical mechanisms of the observed effects, to other non steroidal anti-inflammatory drugs (NSAIDs) (flurbiprofen, indomethacin, acetylsalicylic acid, ibuprofen, nitrofenac and nitroflurbiprofen) and to the effect of high doses diclofenac and flurbiprofen on platelet aggregation. We observed that high doses of diclofenac and of flurbiprofen, but not of the other tested NSAIDs, increased platelet adhesion at doses ranging from 100 to 500 mumol/L, an effect completely removed by the 12-lipoxygenase-inhibitor nordihydroguaiaretic acid. Moreover, they had no pro-aggregating effect, inhibiting platelet aggregation induced by 10 mumol/L arachidonic acid and dose-dependently increasing the [Ca2+]i. Finally, whereas no basal nitric oxide release by washed platelets was detected, when platelets were incubated by 500 mumol/L diclofenac or flurbiprofen, the production of nitric oxide, as measured by amounts of nitrite released, was 4.4 +/- 0.5 and 3.8 +/- 0.4 pmol/5 x 10(8) platelets/min, respectively. Our data indicate that high doses diclofenac and flurbiprofen are promoters of the early phases of platelet activation, probably through the 12-lipoxygenase pathway.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Arachidonate 12-Lipoxygenase; Aristolochic Acids; Calcium; Diclofenac; Dose-Response Relationship, Drug; Enzyme Inhibitors; Flurbiprofen; Humans; In Vitro Techniques; Inflammation; Lipoxygenase Inhibitors; Masoprocol; Nitric Oxide; Phenanthrenes; Phospholipases A; Platelet Activation; Platelet Adhesiveness; Platelet Aggregation

In chronic inflammation, cytokines induce the production of nitric oxide (NO.) that is converted to DNA damaging and carcinogenic peroxynitrite and nitrite. The compounds epigallocatechin gallate (EGCG), carnosol, and curcumin are non-vitamin phytochemicals contained in commonly consumed dietary plants. They are known to be anti-inflammatory and cancer preventive. Therefore, we studied their effect on the generation of peroxynitrite radicals and nitrite. They inhibited lipopolysaccharide (LPS) and interferon-gamma (IFN gamma) induced nitrite production by mouse peritoneal cells by more than 50% at 2.5-10 microM. Cell viability assays verified that the inhibition was not due to general cellular toxicity.

Topics: Abietanes; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Catechin; Cell Survival; Curcumin; Inflammation; Interferon-gamma; Lipopolysaccharides; Macrophage Activation; Macrophages; Mice; Mice, Inbred BALB C; Nitrites; Phenanthrenes; Recombinant Proteins; Spices; Tea; Tumor Necrosis Factor-alpha

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 contribution is concerned with the concept of immunotherapy and the search for the new therapeutic agents. Examples given are, aristolochic acid, Poly I:C and Interferon. The results show a clear correlation of in vivo and in vitro activity of Interferon in the case of virus infections (eccema vaccinatum). No certainty could be determined concerning the efficiency of A.S. and Poly I:C, determined concerning the efficiency of A.S. and Poly I:C, if an Interferon-mechanism was anticipated.

Topics: Aristolochic Acids; Dioxolanes; Humans; Inflammation; Interferons; Leukemia; Neoplasms; Phenanthrenes; Poly I-C; Rosette Formation; Virus Diseases

Topics: Animals; Anti-Inflammatory Agents; Arthritis; Carrageenan; Edema; Formaldehyde; Inflammation; Phenanthrenes; Rats; Saponins

Topics: Humans; Inflammation; Otorhinolaryngologic Diseases; Phenanthrenes

Topics: Abscess; Endocarditis, Bacterial; Humans; Inflammation; Leukocytes; Macrophages; Phagocytosis; Phenanthrenes; Skin Window Technique; Tonsillitis

Topics: Animals; Aristolochic Acids; Communicable Diseases; Drug Resistance; Drug Resistance, Microbial; Infections; Inflammation; Phagocytosis; Phenanthrenes; Plants, Medicinal; Rabbits; Rats; Research; Suppuration