Based On The JAK/STAT Pathway To Explore The Anti-Inflammatory Effect Of Guizhi Shaoyao Zhimu Decoction On TNF-?-Induced MH7A Cells

Objective:This study was designed to investigate the cytotoxicity of Guizhi Shaoyao Zhimu Decoction on Fibroblast-like Synovial cells(MH7A cells)of Rheumatoid Arthritis,and whether Guizhi Shaoyao Zhimu Decoction can inhibit the secretion of inflammatory factors in MH7 A cells induced by tumor necrosis factor-?(TNF-?),and to explore its mechanism.Method:The MH7 A cell line was selected as the experimental research object.Firstly,the effect of Guizhi Shaoyao Zhimu Decoction on the survival rate of MH7 A cells was determined by MTT method,and the appropriate drug concentration was selected.MH7 A cells were divided into blank group,TNF-? group,low-dose group,medium-dose group,and high-dose group.Except the blank group,the other groups were stimulated with TNF-?(10 ng/mL).According to the MTT test results,the drug group was treated with different concentrations of Guizhi Shaoyao Zhimu Decoction(100?g/mL,200?g/mL,600?g/mL),and the cells in each group were further cultured for 24 h.The levels of inflammatory factors(TNF-?,IL-6,IL-1?)in the supernatant fluid of each group were detected by ELISA.The expression of JAK2 and STAT3 protein in each group was detected by Western-blot.Result:1)Guizhi Shaoyao Zhimu Decoction had no obvious cytotoxicity to MH7 A cell line at the concentration below 600 ug/mL.2)Guizhi Shaoyao Zhimu Decoction could inhibit the production of inflammatory cytokines(TNF-?,IL-6,IL-1?)in MH7 A cells induced by TNF-?(P < 0.05).3)The expression of JAK2 protein in the middle and high dose Guizhi Shaoyao Zhimu Decoction group decreased,with no statistical significance(P > 0.05);The expression of STAT3 protein in the low,middle and high dose Guizhi Shaoyao Zhimu Decoction group decreased,with no statistical significance(P > 0.05).Conclusion:The experimental data showed that Guizhi Shaoyao Zhimu Decoction had no obvious cytotoxicity to MH7 A cell line at the concentration below600 ug/mL.Guizhi Shaoyao Zhimu Decoction can reduce the production ofinflammatory factors(TNF-alpha,IL-6,IL-1beta)in MH7 A cells induced by TNF-alpha.Its mechanism may be related to the downward trend of JAK2 and STAT3 protein expression,but its exact mechanisms still need further experimental verification.