OBJECTIVE: To investigate the efficacy of Zhuifeng tougu capsules(追风透骨胶囊, ZFTG) in the treatment of rheumatoid arthritis(RA) in rats and study its mechanism, focusing on the toll-like receptor 2/4-nuclear factor...OBJECTIVE: To investigate the efficacy of Zhuifeng tougu capsules(追风透骨胶囊, ZFTG) in the treatment of rheumatoid arthritis(RA) in rats and study its mechanism, focusing on the toll-like receptor 2/4-nuclear factor kappa-B(TLR2/4-NF-κB) signaling pathway.METHODS: TypeⅡ collagen and an artificial climate box were used to construct the rat model of collagen-induced arthritis with wind-cold-dampness arthralgia syndrome. The rats were divided randomly into a control group, wind-cold-dampness syndrome model group, and high-, medium-,and low-dose ZFTG groups. The methotrexate(MTX) control group was treated with the corresponding drug intervention for 28 d. The joint temperature, pain threshold, joint swelling degree, and arthritis index(AI) score were measured. The production of C-reactive protein(CRP), erythrocyte sedimentation rate(ESR), and rheumatoid factors(RFs) in the blood was detected by enzyme-linked immunosorbent assay. The protein expression of TLR2, TLR4, and NF-κB in synovial tissues was detected by Western blotting, and the m RNA expression of TLR2, TLR4, and NF-κB was detected by real-time polymerase chain reaction.RESULTS: Compared with the model group, the joint temperature in each treatment group, the MTX control group, and MTX group recovered, the degree of foot swelling, pain threshold, AI score decreased, serum CRP, ESR, RF level and the levels of TLR2, TLR4, and NF-κB in synovial tissue were decreased(P < 0.05). Among them, the curative effect in the medium-dose and MTX groups was more evident(P < 0.01).CONCLUSION: ZFTG has a significant effect on RA in rats, and its mechanism may involve regulating CRP levels, the ESR, and RFs via the TLR2/4-NF-κB signaling pathway.展开更多
Objective：To study the mechanism of action of Tougu Xiaotong Capsule（透骨消痛胶囊,TGXTC） ex vivo in suppressing chondrocyte（CD） apoptosis induced by sodium nitroprussiate（SNP）.Methods：Thirty New Zealand rabbit...Objective：To study the mechanism of action of Tougu Xiaotong Capsule（透骨消痛胶囊,TGXTC） ex vivo in suppressing chondrocyte（CD） apoptosis induced by sodium nitroprussiate（SNP）.Methods：Thirty New Zealand rabbits,2 months old,were randomized by lottery into five groups,six in each：the blank group treated with saline,the positive control group treated with Zhuanggu Guanjie Pill（壮骨关节丸,70 mg/kg）,and the three experimental groups,EGA,EGB,and EGC,treated with low dose（35 mg/kg）,moderate dose（70 mg/kg）,and high dose（140 mg/kg） of TGXTC,respectively.All treatments were administered via gastrogavage twice a day for 3 days.Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared.CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining.SNP of various final concentrations（0,0.5,1.0,and 2.0 mmol/L） was used to induce CD apoptosis,and the dosage-effect relationship of SNP in inducing CD apoptosis was determined.Serum samples from the blank,control,and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP.Cell apoptosis was determined by Hoechst 33342 staining,viability of CDs was quantified by MTT,CD apoptosis rate was determined by annexin V-FITC/PI staining,levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR,and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.Results：CD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner.The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study.Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis,decrease in p53 mRNA expression,inhibition of catalytic activities of caspase-3 and caspase-9,and increase in Bcl-2 mRNA expression when compared with the serum from the blank group（P0.05）.Conclusion：TGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression,and inhibition of caspase-3 and caspase-9 catalytic activities.展开更多
Objective： To study the pharmacological properties of Tougu Xiaotong Granule （透骨消痛颗粒, TGXTG） in preventing and treating knee osteoarthritis （KOA） at the molecular level. Methods： The computational methods,...Objective： To study the pharmacological properties of Tougu Xiaotong Granule （透骨消痛颗粒, TGXTG） in preventing and treating knee osteoarthritis （KOA） at the molecular level. Methods： The computational methods, including principal component analysis, molecular docking, target-ligand space distribution, and the predictions of absorption, distribution, metabolism, excretion and toxicity （ADMET）, were introduced to characterize the molecules in TGXTG. Results： The structural properties of molecules in TGXTG were more diverse than those of the drug/drug-like molecules, and TGXTG could interact with significant target enzymes related to KOA. In addition, the cluster of effective components was preliminarily identified by the target-ligand space distributions. As to the results of ADMET properties, some of them were unsatisfactory, and were merely regarded as references here. Conclusion： Based on this computational pharmacology study, TGXTG is a broad- spectrum recipe inhibiting many important target enzymes, which could effectively postpone the degeneration of cartilage by coordinately inhibiting the biological effects of cytokines, matrix metallopeptidase 3, and oxygen free radicals.展开更多
基金Supported by Natural Science Foundation of Hunan Province of China (the Mechanism of Zhuifengtougu Capaule on the Cold and Heat Syndrome Animal Model of Reheumatod Arthritis in Rats on the "Syndrome-Molecular" No.2017JJ2190Research the Effect Mechanism of Zhuifengtougu Capsule on the Knee Osteoarthritis Base on TLR4-My D88Pathway Mediated by Intestinal Microbiome, 2019JJ50462)。
文摘OBJECTIVE: To investigate the efficacy of Zhuifeng tougu capsules(追风透骨胶囊, ZFTG) in the treatment of rheumatoid arthritis(RA) in rats and study its mechanism, focusing on the toll-like receptor 2/4-nuclear factor kappa-B(TLR2/4-NF-κB) signaling pathway.METHODS: TypeⅡ collagen and an artificial climate box were used to construct the rat model of collagen-induced arthritis with wind-cold-dampness arthralgia syndrome. The rats were divided randomly into a control group, wind-cold-dampness syndrome model group, and high-, medium-,and low-dose ZFTG groups. The methotrexate(MTX) control group was treated with the corresponding drug intervention for 28 d. The joint temperature, pain threshold, joint swelling degree, and arthritis index(AI) score were measured. The production of C-reactive protein(CRP), erythrocyte sedimentation rate(ESR), and rheumatoid factors(RFs) in the blood was detected by enzyme-linked immunosorbent assay. The protein expression of TLR2, TLR4, and NF-κB in synovial tissues was detected by Western blotting, and the m RNA expression of TLR2, TLR4, and NF-κB was detected by real-time polymerase chain reaction.RESULTS: Compared with the model group, the joint temperature in each treatment group, the MTX control group, and MTX group recovered, the degree of foot swelling, pain threshold, AI score decreased, serum CRP, ESR, RF level and the levels of TLR2, TLR4, and NF-κB in synovial tissue were decreased(P < 0.05). Among them, the curative effect in the medium-dose and MTX groups was more evident(P < 0.01).CONCLUSION: ZFTG has a significant effect on RA in rats, and its mechanism may involve regulating CRP levels, the ESR, and RFs via the TLR2/4-NF-κB signaling pathway.
文摘Objective：To study the mechanism of action of Tougu Xiaotong Capsule（透骨消痛胶囊,TGXTC） ex vivo in suppressing chondrocyte（CD） apoptosis induced by sodium nitroprussiate（SNP）.Methods：Thirty New Zealand rabbits,2 months old,were randomized by lottery into five groups,six in each：the blank group treated with saline,the positive control group treated with Zhuanggu Guanjie Pill（壮骨关节丸,70 mg/kg）,and the three experimental groups,EGA,EGB,and EGC,treated with low dose（35 mg/kg）,moderate dose（70 mg/kg）,and high dose（140 mg/kg） of TGXTC,respectively.All treatments were administered via gastrogavage twice a day for 3 days.Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared.CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining.SNP of various final concentrations（0,0.5,1.0,and 2.0 mmol/L） was used to induce CD apoptosis,and the dosage-effect relationship of SNP in inducing CD apoptosis was determined.Serum samples from the blank,control,and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP.Cell apoptosis was determined by Hoechst 33342 staining,viability of CDs was quantified by MTT,CD apoptosis rate was determined by annexin V-FITC/PI staining,levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR,and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.Results：CD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner.The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study.Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis,decrease in p53 mRNA expression,inhibition of catalytic activities of caspase-3 and caspase-9,and increase in Bcl-2 mRNA expression when compared with the serum from the blank group（P0.05）.Conclusion：TGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression,and inhibition of caspase-3 and caspase-9 catalytic activities.
基金Supported by the National Natural Science Foundation of China (No.30672701)Chen Ke-ji Integrative Medicine Development Foundation(No.2008J1004-27CKJ2008064)Major Project of Department of Science & Technology,Fujian Province(No. 2006Y0016,2009Y0029)
文摘Objective： To study the pharmacological properties of Tougu Xiaotong Granule （透骨消痛颗粒, TGXTG） in preventing and treating knee osteoarthritis （KOA） at the molecular level. Methods： The computational methods, including principal component analysis, molecular docking, target-ligand space distribution, and the predictions of absorption, distribution, metabolism, excretion and toxicity （ADMET）, were introduced to characterize the molecules in TGXTG. Results： The structural properties of molecules in TGXTG were more diverse than those of the drug/drug-like molecules, and TGXTG could interact with significant target enzymes related to KOA. In addition, the cluster of effective components was preliminarily identified by the target-ligand space distributions. As to the results of ADMET properties, some of them were unsatisfactory, and were merely regarded as references here. Conclusion： Based on this computational pharmacology study, TGXTG is a broad- spectrum recipe inhibiting many important target enzymes, which could effectively postpone the degeneration of cartilage by coordinately inhibiting the biological effects of cytokines, matrix metallopeptidase 3, and oxygen free radicals.
