茵芪三黄解毒汤抑制中性粒细胞-胞外诱捕网的抗肝纤维化作用机制OA

Objective:To verify the therapeutic effect of Yinqi Sanhuang Jiedu decoction(YQSH)on carbon tetrachloride(CCl4)-induced hepatic fibrosis in mice,and to explore whether its effect was related to the inhibition of neutrophil infiltration and the formation of neutrophil extracellular traps(NETs).Methods:The 36 C57BL/6J mice were randomly divided into control group,model group,positive drug silybin(SF)group(55 mg·kg-1·d-1),YQSH-L group,YQSH-M group,and YQSH-H group(8.325,16.65,33.3 g·kg-1·d-1,respectively),n=6 in each group.Except for the control group,mice in all other groups were intraperitoneally injected with CCl4 to induce hepatic fibrosis.After successful modeling,each drug administration group was given the corresponding drugs by gavage for eight weeks.Hematoxylin-eosin(HE)staining,Sirius red staining and Masson staining were used to observe the pathological changes of liver tissue.Liver elasticity was detected by a color Doppler ultrasound system.Immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)were performed to detect the protein expression and mRNA levels of C-X-C motif chemokine ligand 1(CXCL1),CXCL2 and CXCL5.Neutrophil levels were detected by flow cytometry.The expression of neutrophil elastase(NE)and myeloperoxidase(MPO)positive protein was observed by immunofluorescence.The contents of MPO,NE and CitH3 were detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the control group,the liver of the model group showed obvious inflammatory cell infiltration and collagen deposition,and the liver elasticity,CXCL1,CXCL2,CXCL5 expression,neutrophil level,and MPO,NE and CitH3 levels were significantly increased(P＜0.05,P＜0.01).Compared with the model group,inflammatory cell infiltration and collagen deposition in the liver tissue of mice were reduced after YQSH treatment.Moreover,the liver elasticity was reduced(P＜0.01).The protein expression(P＜0.01)and mRNA level of CXCL1,CXCL2 and CXCL5 were decreased(P＜0.05,P＜0.01).The neutrophil level was decreased(P＜0.01),the expression of MPO and NE positive protein was significantly decreased(P＜0.05,P＜0.01),and the levels of MPO,NE and CitH3 were decreased(P＜0.05,P＜0.01).Conclusion:The anti-hepatic fibrosis effect of YQSH may be related to its inhibition of chemokines(CXCL1,CXCL2,CXCL5),reduction of neutrophil infiltration,and inhibition of NETs generation.