Mechanism of Buyang Huanwu decoction in treating non-alcoholic fatty liver disease based on network pharmacology and experimental validation

AimTo screen the therapeutic mechanisms of Buyang Huanwu decoction (BYHWD) in treating non-alcoholic fatty liver disease (NAFLD) based on network pharmacology and to validate them through cell experiments.MethodsThe active components and potential targets of BYHWD in the treatment of NAFLD were obtained through relevant databases of network pharmacology, and the"drug–component–target"relationship network was constructed. GO and KEGG enrichment analysis was performed on the intersecting genes. L02 cells were cultivated in vitro and an NAFLD model was established by inducing L02 cells with oleic acid(oleic acid, OA). The effects of different concentrations of DMSO, OA, and BYHWD on the viability of L02 cells were analyzed using the CCK-8 assay. The intracellular TG content was detected using a triglyceride assay kit. Intracellular lipid accumulation was observed by Oil Red O staining. The effects of BYHWD on lipid content and oxidative stress injury in OA-induced L02 cells were detected using biochemical methods. The changes in mRNA and protein levels of AMPK and SREBP-1c, GPAT were detected using real-time quantitative PCR (RT-qPCR) and Western blot methods.ResultsA total of 111 active components of BYHWD were screened out, involving 2 264 targets, among which 531 intersecting targets were shared with NAFLD. The KEGG analysis results showed that the treatment of NAFLD by BYHWD was mainly enriched in signaling pathways such as lipid and atherosclerosis, AMPK signaling pathway, and insulin resistance. The cellular results indicated that BYHWD could significantly reduce the levels of TG(triglycerides)and MDA(malondialdehyde), inhibit the activity of AST (aspartate aminotransferase), and enhance the activity of SOD (superoxide dismutase). Moreover, BYHWD promoted the expression of AMPK at both gene and protein levels, while inhibiting the expression of SREBP- 1c(sterol regulatory element-binding protein 1c)and GPAT (glycerol-3-phosphate acyltransferase) at both gene and protein levels.ConclusionBYHWD may treat NAFLD by regulating the AMPK and SREBP-1c, GPAT signaling pathways, reducing lipid synthesis and accumulation, thereby ameliorating oxidative stress injury.