Study On The Lipid-lowering Effect Of Artemisinin And Its Derivatives In Vitro

Objective: Artemisinin and its derivatives have been reported to play an important role in treating cardiovascular diseases such as atherosclerosis,obesity and diabetes,possibly by anti-inflammatory and lipid-lowering.This study intends to use cell models to study the anti-inflammatory and lipidlowering effects of artemisinin and its derivatives,and to carry out a series of preliminary researches on its mechanism of action,providing a theoretical basis for its application in cardiovascular diseases.Methods: In this study,we used LPS to induce the mouse macrophages RAW264.7 to an inflammatory model and cholesterol-induced RAW264.7 and FFA-induced human liver cancer cells HepG2 to establish high-fat models.Three cell models were used to evaluate the anti-inflammatory and lipid-lowering effects of artemisinin and its derivatives.Transcriptome sequencing and qRT-PCR validation were used to explore the mechanism of lipid-lowering in artemisinin compounds.Results: Artesunate and artemisinin can significantly reduce the accumulation of lipids in HepG2 cells induced by FFA.Artemisinin can significantly reduce the accumulation of lipids in RAW264.7 cells induced by cholesterol.However,artesunate in this concentration range up-regulated the expression of TNF-? and IL-6 in RAW264.7 cells.Which indicated that the effect of artemisinin compounds on lipid metabolism was independent of its anti-inflammatory mechanism.The above results indicated that the HepG2 high-fat model was more suitable for the regulation of lipid metabolism by artemisinin compounds.The transcriptome sequencing data of artesunate-treated HepG2 cells showed that there were 2679 differentially significant genes in the artesunate treated group compared with the model group,all of which were annotated into 61 GO secondary items and 305 KEGG signaling pathways.A total of 14 lipid metabolic signaling pathways have been annotated,which are related to fatty acid metabolism and there are 20 metabolic pathways related to FFA-induced HepG2 fatty liver in vitro.Among these pathways,44 and 22 distinctly differential genes were annotated into the AMPK and PPAR signaling pathways,including the core genes PRKAA and PPARG and their upstream and downstream genes.qRT-PCR experiments confirmed that the artesunate treated group significantly increased the mRNA levels of PRKAA and PPARG genes compared with the model group.Conclusion: Artemisinin can significantly reduce the phagocytosis of exogenous cholesterol in RAW264.7 cells.Artemisinin,rFIP-gat and artesunate can significantly reduce the accumulation of FFA in HepG2 cells.The lipid-lowering effect of artesunate may up-regulate the expression of PRKAA and PPARG to mediate AMPK and PPAR signaling pathways.