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 dismutase （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.

Key words: Qushi Huoxue decoction, non-alcoholic fatty liver disease, AMPK/SREBP1c signaling pathway, HepG2 cells