The effect of the Qushi Huoxue decoction on the AMPK/SREBP1c signaling pathway in free fatty acid-induced HepG2 cells

TAN Weiqiang; LIU Li; LIU Xudong; WEI Yehuang; GONG Xiaoqian; WANG Rongrong

doi:10.3969/j.issn.1006-5725.2026.03.007

Abstract：Objective To investigate the effect of Qushi Huoxue decoction(QSHXF)on free fatty acid(FFA)-induced HepG2 cells and its underlying mechanism.Methods In vitro cell experiments were conducted.First,the CCK-8 assay was used to determine the effects of different concentrations of QSHXF on HepG2 cells viability to select appropriate concentrations for subsequent experiments.A fatty liver cell model was then established by treating HepG2 cells with a mixture of 250 μmol/L sodium palmitate and 500 μmol/L sodium oleate for 24 h,followed by intervention with different concentrations of QSHXF-containing serum and AMPK inhibitor for another 24 h.Intracellular lipid droplet accumulation was observed using Oil Red O staining and lipid fluorescence staining.The levels of total cholesterol(TC)and triglycerides(TG)were measured using the GPO-PAP enzymatic method.The TBA method and hydroxylamine method were used to detect malondialdehyde(MDA)and superoxide dis-mutase(SOD),respectively,while ELISA was used to detect tumor necrosis factor-α(TNF-α)and interleukin 1β(IL-1β).Furthermore,qRT-PCR,Western blot,and immunofluorescence staining were performed to detect the expression of lipid metabolism-related genes and proteins.Results QSHXF significantly improved the lipid deposition,oxidative stress and inflammatory response of FFA-induced HepG2 cells,markedly reduce the content of TC,TG,MDA,TNF-α and IL-1β in cells and increase the activity of SOD(P<0.01).Meanwhile,it increased the mRNA expression of AMPK and the protein ratio of p-AMPK/AMPK,while decreasing the mRNA and protein levels of SREBP1c,FASN,ACC1,and SCD1(all P<0.05).Conclusion QSHXF significantly improved lipid metabolism and alleviated oxidative stress and inflammation in the MASLD cell model,and its mechanism may be associated with the activation of the AMPK/SREBP1c signaling pathway.