A valproic acid-induced cell model of hepatic steatosis: Effects of <i>Gnetum montanum</i> extract

Hepatic steatosis is a multifactorial disorder with no specific pharmacological treatment available. Gnetum montanum has been traditionally used to treat metabolic disorders; however, its effects on hepatic steatogenesis remain unexplored. This study aimed to establish a method for inducing steatosis in hepatocytes and to investigate whether G. montanum extract (GME) exerts beneficial effects on hepatocellular lipid metabolism. We found that valproic acid (VPA) at a concentration of 3 mM effectively induced extensive lipid accumulation in HepG2 human hepatoma cells, as evidenced by the formation of intracellular lipid droplets and an increase in cellular triglyceride and cholesterol levels, with minimal effects on cell viability after 48 hours of incubation. Interestingly, pretreatment with GME significantly abrogated the steatogenic effects of VPA. Specifically, GME reduced the intracellular neutral lipid pool, as determined by Oil Red O staining. Fluorescent assays using BODIPY 493/503 in combination with imaging techniques and flow cytometry further confirmed that GME decreased both the number of lipid droplets and the accumulation of neutral lipids in VPA-treated HepG2 cells. Additionally, GME exerted dose-dependent inhibitory effects on VPA-induced increases in cellular triglyceride and total cholesterol levels. Taken together, this study has established a cell model for evaluating in vitro anti-hepatic steatotic activity and suggests G. montanum as a promising medicinal plant for managing hepatic steatosis. Keywords: Gnetum montanum, Hepatic steatosis, Lipid droplets, NALFD, Valproate.