Wenjing Huoluo Recipe Treated Collagen-induced Arthritis Model Mice
Objective To study the therapeutic mechanisms for Wenjing Huoluo Recipe(WHR)in treatment of collagen-induced arthritis(CIA)via inhibition of tumor necrosis factor-α(TNF-α)/nuclear factor kappa B(NF-κ B)pathway and regulation of Th17 cells in a murine model.Methods Forty 6-week male DBA/1 mice were randomly divided into four groups:normal control group,model group,methotrexate(MTX)group,and WHR group,10 in each group.CIA model was established by collagen injection,and intervened with double distilled water,MTX,and WHR for 35 days starting from the 21st day after initial immunization.Body weight of mice in each group was assessed weekly.The degree of foot swelling and joint inflammation score of each group were evaluated weekly from the 24th day.HE staining was used to detect pathological changes in the toes and ankle joints.ELISA was used for detecting anti-cyclic citrullinated peptide(CCP)antibodies,serum TNF-α and IL-6 levels.Real-time quantitative polymerase(RT-qPCR)was used to detect mRNA expression levels of foot joint NF-κ B,osteoprotegerin(OPG),and receptor activator of nuclear factor-κ B ligand(RANKL).Western Blot was used to detect NF-κB and P-NF-κB expressions in mouse spleen and foot joints.Flow cytometry was used to detect the expression of helper T lymphocyte 17(Th17)cells in the mouse spleen.Results Compared with the normal group,the hair withered,food intake reduced,and body weight obviously decreased in the model group(P＜0.05,P＜0.01).Contents of anti-CCP antibody,IL-6,and TNF-α increased(P＜0.01).mRNA expressions of NF-κB and RANKL also increased(P＜0.01),OPG mRNA expression obviously decreased(P＜0.05).p-NF-κ B protein expression obviously increased in the spleen and toe joints(P＜0.01).Th17 cell ratio in the spleen significantly increased(P＜0.01).Compared with the model group,the weight loss in WHR was alleviated(P＜0.05).The swollen spleen was significantly alleviated in each treatment group(P＜0.01).The degree of foot swelling was significantly reduced(P＜0.01).The joint inflammation score significantly decreased in mice of WHR group(P＜0.05,P＜0.01).The proliferation of synovial cells in the toes and ankle joints of mice was attenuated,and the number of infiltrated inflammatory cells was significantly reduced,the formation of pannus decreased,and the destruction of bone and cartilage were significantly attenuated after treated by WHR.After drug intervention the levels of serum anti-CCP antibodies,IL-6,and TNF-α in each treatment group decreased more significantly(P＜0.05,P＜0.01).After drug treatment NF-κ B phosphorylation(p-NF-κB)level decreased(P＜0.05,P＜0.01),and mRNA expression levels of NF-κ B,and RANKL significantly decreased(P＜0.01),the mRNA expression level of OPG significantly increased(P＜0.05,P＜0.01),the expression of Th17 cells was significantly inhibited after treatment with MTX and WHR(P＜0.01).Conclusion WHR inhibited TNF-α/NF-κ B signaling pathway and Th17 cells'activation possibly by further inhibiting expressions of TNF-α,IL-6 and other related inflammatory factors,thus attenuating RA joint inflammation,lowering joint inflammation score inhibiting bone destruction,and improving pathological changes.
国家科技期刊平台
NSTL
万方数据
