Exploration of the mechanism of Alisol B in improving NAFLD of L-02 cells under hypoxic through the PI3K/AKT-mTOR-HIF1α signaling pathway

Abstract Objective: This study aims to explore the mechanism by which the Alisol B exerts its lipid-regulating effects on non-alcoholic fatty liver disease (NAFLD) and to investigate the impact of a hypoxic environment on the progression of NAFLD. Methods: Oil red O staining and lipid fluorescence staining were used to determine the lipid accumulation in L-02 cells; oxidative stress and liver lipid metabolism biochemical indexes were detected in the cell samples by using kits; network pharmacology, molecular docking and other techniques were used to predict the relevant targets for the lipid-lowering and antioxidant effects exerted by Alisol B, and the intersection of the targets were summarized and analyzed by GO and KEGG pathway enrichment. Western blot and qPCR were applied to detect the relative expression of the relevant proteins and target genes. Results: Alisol B can effectively regulate the four abnormalities of blood lipids triggered by NAFLD and improve the up-regulation of liver injury indexes caused by disease and hypoxic environment. Through the network pharmacology technology, 21 related gene targets were screened, 240 entries of GO function enrichment analysis were obtained, and 127 signaling pathways of KEGG pathway enrichment analysis were analyzed. Further experiments demonstrated that the inhibition of the PI3K/AKT-mTOR-HIF1α pathway can down-regulate the expression of associated lipid synthesis genes, while lipid metabolism genes exhibit a reverse regulatory effect. Simultaneously, the disease exacerbation induced by a hypoxic environment was alleviated. In conclusion, Alisol B protects the liver by inhibiting the PI3K/AKT-mTOR-HIF1α signaling pathway, thereby regulating lipid-related genes and improving lipid metabolism dysfunction in a hypoxia-induced disease model. Conclusion: Alisol B protects the liver by inhibiting the PI3K/AKT-mTOR-HIF1α signaling pathway, which regulates lipid-related genes and improves lipid metabolism dysfunction in disease models induced by hypoxia.