祛湿活血方对非酒精性脂肪肝肝细胞焦亡诱导巨噬细胞活化的影响OA

Objective To explore the mechanism of Qushi Huoxue Decoction(祛湿活血方,QSHXF)in treating non-alcohol-ic steatohepatitis(NASH)from the perspective of activation of macrophages induced by pyroptosis of hepatocytes in non-alcohol-ic fatty liver disease(NAFLD).Methods The high and low concentrations of serum containing QSHXF were used to intervene the pyroptosis of HepG2 cells induced by lipopolysaccharide(LPS).Adiponectin(ADPN)was used as a positive drug control group.The cell proliferation rate was determined using the CCK-8 method.The concentrations of interleukin-18(IL-18),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in cell culture supernatant were detected by ELISA.The mRNA expres-sions of CASP1 and GSDMD were detected by real-time quantitative PCR.The expressions of pyroptosis-related proteins were detected by Western blot.The effect of QSHXF on the recruitment of J774A.1 by HepG2 was observed by Transwell culture meth-od.At the same time,the secretion of inflammatory factors after activation of J774A.1 induced by HepG2 was detected.Results After 24 and 48 hours of LPS intervention,the proliferation rate of HepG2 cells in the model group was significantly higher than that in the QSHXF-H group(P＜0.01).The levels of TNF-α,IL-18 and IL-1β in the ADPN group and the QSHXF-H group were lower than those in the model group(P＜0.05).At the same time,the levels of IL-18 and IL-1β in the QSHXF-L group ere lower than those in the model group(P＜0.05).The expression of CASP1 mRNA in the QSHXF-H group was sig-nificantly lower than that in the model group(P＜0.05).The expressions of GSDMD mRNA in the ADPN group and the QSHXF-L group were lower than that in the model group(P＜0.05).The expressions of NLRP3,CASP1,GSDMD and GSDMD-N pro-teins in the ADPN group and the QSHXF-H group were all lower than that in the model group(P＜0.01).The content of GSD-MD in HepG2 cells in the QSHXF-L group was lower than that in the model group(P＜0.01).The gray values of CASP1 in the ADPN group,QSHXF-H group and QSHXF-L group were lower than those in the model group(P＜0.01).In the co-culture experiment,the levels of IL-1,IL-1β,IL-6,IL-10,IL-18 and TNF-α in the ADPN group and the QSHXF groups were significantly lower than those in the model group(P＜0.05).The numbers of J774A.1 cells in the ADPN group and the QSHXF groups were significantly higher than that in the model group(P＜0.01).Conclusion QSHXF may inhibit the pyroptosis of hepa-tocytes,thus reducing the recruitment of macrophages,the activation of macrophages and the secretion of inflammatory factors,and then treating NASH.