Protective Effect and Mechanism of Bitong Decoction(痹通方)on Rheumatoid Arthritis Rats Based on NLRP3 Inflammasome Mediated Pyroptosis
Objective To investigate the protective effect and mechanism of Bitong Decoction(痹通方)on rheumatoid arthritis rats by regulating the pyroptosis pathway mediated by NLRP3 inflammasome.Methods Seventy SD rats were randomly divided in-to 7 groups:Bitong Decoction low-dose,medium-dose and high-dose groups[4.2,8.4,16.8 g/(kg·d)by gavage],metho-trexate group[7.5 mg/(kg·d)by gavage],MCC950 group[30 mg/(kg·d)by gavage].The normal group and model group were given distilled water by gavage.On the 29th day after modeling,the drug was administered by gavage once a day.HE stai-ning was used to detect the morphological changes of ankle joints in each group at 8 h after the last administration.The changes of tumor necrosis factor-α(TNF-α),interleukin-18(IL-18)and interleukin-1β(IL-1β)related indexes were detected by ELISA.The levels of TNF-α,IL-18,IL-1β,monocyte chemotactic protein-1(MCP-1)and macrophage inflammatory protein-1a(MIP-1a)related genes were detected by RT-qPCR.The protein levels of IL-1 β and NOD like receptor heat protein domain associated protein 3(NLRP3)were detected by immunohistochemistry.Melanoma deficiency factor 2(AIM2),ap-optosis related point like protein(ASC),cysteine protease precursor 1(pro Caspase-1),Caspase-1,IL-18,IL-1 β,cleaved-GSDMD,GSDMD and NLRP3 were detected by Western Blot.Results Compared with those of the normal group,the ankle joint space of the model group was narrowed,the surface of articular cartilage was irregularly thickened,the bone was slightly de-stroyed,the inflammatory cells and synovial tissue hyperplasia were observed in the joint(P＜0.01),and the serum contents of TNF-α,IL-18 and IL-1 β in the model group were significantly increased(P＜0.01).The mRNA expressions of TNF-α,IL-18,IL-1 β,MCP-1 and MIP-1a in the model group were significantly increased(P＜0.01).Immunohistochemistry showed that the protein expressions of IL-1β and NLRP3 in the model group were significantly increased(P＜0.01).Western Blot method showed AIM2,ASC,pro-Caspase-1,Caspase-1,IL-18 and IL-1 β in the model group were significantly in-creased(P＜0.01).Compared with those of the model group,the above indexes in the Bitong Decoction high-dose group,meth-otrexate group and MCC950 group were significantly improved(P＜0.05 or P＜0.01).Conclusion Bitong Decoction can improve the state of rheumatoid arthritis in rats,and its mechanism may be related to the regulation of NLRP3 inflammasome mediated py-roptosis pathway.
万方数据
维普
