Effect of Alisol G on Lipid Metabolism in HepG2 Cells Fat Accumulation Model Using Mass Spectrometry Based Lipidomics

OBJECTIVE To explore the mechanism of alisol G(AG) in improving lipid metabolism disorder in HepG2 cells fat accumulation model using lipidomics.

METHODS HepG2 cells fat accumulation model was constructed using free fatty acids solution(oleic acid∶palmitic acid=2∶1). After intervention with AG, oil red O staining was performed, and the relative lipid content, triglycerides(TG) content, and total cholesterol(TC) content in the cells were measured, respectively. Ultra-high performance liquid chromatography-quadrupole time of flight-mass spectrometry/mass spectrometry(UPLC-QTOF-MS/MS) techniques was applied to analyze the changes in the cellular lipid composition and the effects of lipid metabolism pathways. The mechanism of the effect of AG on lipid metabolism of HepG2 cells was predicted by network pharmacology, and the binding ability of AG to key targets was verified by molecular docking technology.

RESULTS 5 µmol·L⁻¹ and 10 µmol·L⁻¹ of AG significantly inhibited the lipid content in the FFA induced HepG2 cells fat accumulation model(P<0.05 or P<0.01), and significantly reduced the levels of TG and TC(P<0.01). Sixteen potential differential lipid metabolites including glycerophospholipids, sphingolipids, and sterols, were identified through lipidomics analysis, which mainly affect the process of fat accumulation in HepG2 cells through the glycerophospholipid metabolic pathway. Network pharmacologic analysis identified PPARG, PPARA, ESR1, AKT1, AKT2, PIK3CA and other key targets, suggesting that these targets played a key role in the improvement of lipid metabolism disorders by AG. Molecular docking results showed that the lowest binding energy between AG and these key targets was −6.0 kcal·mol⁻¹, indicating that AG had high affinity with key targets and high drug activity.

CONCLUSION AG may primarily influence the glycerophospholipid pathway by regulating glycerophospholipids, sphingolipids, and sterol lipids; it may mainly act on targets such as AKT1, AKT2, PPARG and PPARA to affect multiple signaling pathways including PI3K-AKT, thereby exerting the potential to regulate lipid metabolism and improve fat accumulation in HepG2 cell fat accumulation models.