Effect of regulating miR-29b-3p/FoxO3 axis by asiaticoside on alleviating liver injury in rats with non-alcoholic fatty liver disease

TAN Jie; LIU Meng; TIAN Xia; HAN Zheng; ZHU Qingxi; ZHAN Ting; CHEN Mingtao; CHEN Aifang; HUANG Min

doi:10.12173/j.issn.1004-5511.202501068

Abstract：Objective To investigate the effect of Asiaticoside(ASI)on liver injury in non-alcoholic fatty liver disease(NAFLD)and its impacts on the microRNA-29b-3p(miR-29b-3p)/forkhead box protein O3(FoxO3)axis.Methods Rats were stochastically grouped into a Control group,a Model group,a low-dose ASI-L group(ASI-L),a high-dose ASI-H group(ASI-H),a high-dose ASI+blank vector group(ASI-H+Vector),and a high-dose ASI+miR-29b-3p over expression group(ASI-H+OV-miR-29b-3p),with 12 rats in each group.The changes in rat liver index were analyzed.HE staining method was applied to observe the pathological changes of liver tissue.Biochemical analyzer was used to detect lipid metabolism indicators and indicators reflecting liver parenchymal damage in rats.The levels of oxidative stress and inflammatory cytokines were detected by the ELISA method.The RT-qPCR method was applied to detect the mRNA levels of miR-29b-3p and FoxO3 in rat liver tissue.The Western blot method was applied to detect the expression levels of FoxO3 and apoptosis-related proteins in rat liver tissue.Dual luciferase activity was applied to validate the targeting relationship between miR-29b-3p and FoxO3.Results Compared with the Control group,rats in the Model group showed severe liver tissue damage,liver cell rupture,and extensive infiltration of inflammatory cells,the liver weight and liver index increased,the levels of TC,TG,AST,ALT,MDA,TNF-α,IL-6 obviously increased,the levels of SOD and CAT obviously reduced,the expression level of miR-29b-3p and expression level of Caspase 3 protein in liver tissue obviously increased,the expression level of FoxO3 mRNA and expression levels of FoxO3,Bcl-2 proteins decreased(P＜0.05).Compared with the model group,as the dose of ASI increased,the degree of liver tissue damage,liver cell rupture,and inflammatory cell infiltration of rats in the ASI-L and ASI-H groups decreased,the liver weight and liver index gradually decreased,the levels of TC,TG,AST,ALT,MDA,TNF-α,IL-6 in serum gradually decreased,the levels of SOD and CAT gradually increased,the expression level of miR-29b-3p and expression level of Caspase 3 protein in liver tissue gradually decreased,the expression level of FoxO3 mRNA and the expression levels of FoxO3 and Bcl-2 proteins gradually increased.miR-29b-3p overexpression reversed the improvement effect of ASI on liver tissue in NAFLD rats.Dual luciferase activity confirmed the targeting relationship between miR-29b-3p and FoxO3.Conclusion ASI can enhance the antioxidant function,regulate lipid metabolism,alleviate inflammation and liver tissue damage in NAFLD rats,which may be related to the down-regulation of miR-29b-3p and the up-regulation of FoxO3 expression.