Integrated Multi-Omics Investigation of Gypenosides’ Mechanisms in Lowering Hepatic Cholesterol

(1) Objective: This study aimed to systematically elucidate the molecular mechanisms by which gypenosides (GP), a major active component of <i>Gynostemma pentaphyllum</i>, ameliorate hypercholesterolemia by modulating the hepatic steroidogenesis pathway, and to identify key therapeutic targets. (2) Methods: We established a high-fat diet (HFD)-induced hypercholesterolemia (HC) mouse model and performed GP intervention. An integrated multi-omics approach, combining transcriptomics and proteomics, was utilized to comprehensively analyze GP's effects on the expression of genes and proteins associated with hepatic cholesterol synthesis, transport, and steroid hormone metabolism. (3) Results: HFD induced significant dysregulation, with 48 steroidogenesis pathway-related genes and 35 corresponding proteins exhibiting altered expression in HC mouse livers. GP treatment remarkably reversed these HFD-induced abnormalities, significantly restoring the expression levels of 42 genes and 14 proteins. Multi-omics integration identified seven critical genes/proteins-<i>Cyp3a25</i>, <i>Fdft1</i>, <i>Tm7sf2</i>, <i>Hmgcs1</i>, <i>Fdps</i>, <i>Mvd</i>, and <i>Pmvk</i>-that were consistently and significantly regulated by GP at both transcriptional and translational levels. Furthermore, correlation analyses demonstrated that <i>Cyp3a25</i> was significantly negatively correlated with serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), whereas <i>Fdft1</i>, <i>Tm7sf2</i>, <i>Hmgcs1</i>, <i>Fdps</i>, <i>Mvd</i>, and <i>Pmvk</i> showed significant positive correlations. (4) Conclusions: GP effectively ameliorates cholesterol dyshomeostasis through a multi-targeted mechanism in the liver. It inhibits endogenous cholesterol synthesis by downregulating key enzymes (<i>Hmgcs1</i>, <i>Fdft1</i>, <i>Pmvk</i>, <i>Mvd</i>, <i>Fdps</i>, <i>Tm7sf2</i>), promotes cholesterol efflux and transport (upregulating <i>Abca1</i>, <i>ApoB</i>), and accelerates steroid hormone metabolism (upregulating <i>Cyp3a11</i>, <i>Cyp3a25</i>). These findings provide robust scientific evidence for the development of GP as a safe and effective novel therapeutic agent for hypercholesterolemia.