Protective Effect and Mechanism of Serum Containing Bitong Decoction(痹通方)on Fibroblast-Like Synovial Cell Injury in Rheumatoid Arthritis Based on NLRP3 Mediated Pyroptosis
Objective To investigate the protective mechanism of Bitong Decoction(痹通方)containing serum on pyroptosis of fibroblast-like synoviocytes(MH7A)induced by lipopolysaccharide(LPS)and its effect on NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway in rheumatoid arthritis.Methods SD rats were randomly divided into 3 groups:control group,Bitong Decoction low and high dose groups[4.2 g/(kg·d)and 16.8 g/(kg·d)by gavage,respectively].The control group was given the same volume of distilled water by gavage.The drug-containing serum was prepared after 10 days of continuous intragastric administration.The cells were divided into 5 groups:control group,LPS treatment group,low and high dose serum containing Bitong Decoction group,MCC950 group.The cells in the above groups were collected,and the cell viability was detected by CCK8.The expression levels of tumor necrosis factor-α(TNF-α),interleukin-18(IL-18),interleukin-1 β(IL-1β),monocyte chemotactic protein-1(MCP-1)and macrophage inflammatory protein 1 α(MIP-1α)were detected by RT-qPCR.The protein expressions of AIM2,ASC,Caspase-1,IL-18,IL-1 β,cleaved-GSDMD,GSDMD and NLRP3 were detected by Western blotting.The expression of NLRP3 was detected by immunofluorescence staining.Results Compared with that of the control group,the viability of cells in LPS treatment group was decreased(P＜0.01),and the mRNA expressions of TNF-α,IL-18,IL-1 β,MCP-1 and MIP-1A were increased(P＜0.01).The protein expressions of AIM2,ASC,Caspase-1,IL-18,IL-1 β,cleaved-GSDMD,GSDMD and NLRP3 were significantly increased(P＜0.01).The results of immunofluo-rescence showed that the fluorescence intensity of NLRP3 in the model group was significantly higher than that in the blank con-trol group(P＜0.01).Compared with LPS treatment group,NLRP3 inhibitor MCC950 group and serum containing Bitong Decoc-tion groups could improve the changes of above indexes.Conclusion Bitong Decoction can effectively inhibit the expression of NLRP3 inflammasome,reduce pyroptosis of MH7A cells and improve inflammatory injury.Its mechanism may be related to the regulation of NLRP3 pathway.
rheumatoid arthritisMH7A cellspyroptosis of cellsBitong Decoction(痹通方)NLRP3
万方数据
维普
