COX-2Regulates Acsl Family Genes Expression In Palmitic Acid And Oleic Acid Induced Hepatic Stellate Cells Re-lipidation

Aim：This chapter mainly discussed the effect of palmitic acid and oleic acid onHSC-T6re-lipidation, and revealed the regulation effect of COX-2on the expressionof long-chain acyl-AcylCoA synthetase（Acsl）during palmitic acid and oleic acidinduced rat hepatic stellate cells (HSC-T6) re-lipidation.Methods:1.The re-lipidation of HSC-T6induced by DMEM substrate which contained200mmol/L palmitic acid or20μg/ml oleic acid. Test the variation of lipid drops inHSC-T6treatment with youhong O staining48h, to exam the effect of palmitic acidand oleic acid on re-lipidation.2. Transfer the constructed pYr-1.1-COX-2shRNA into rat HSC-T6, then assess thetransfection efficiency under the fluorescence microscope.3.Western Blot: detect the expression of COX-2, Acsl1, Acsl3, Acsl4, Acsl5, Acsl6after transfer COX-2shRNA for48h by palmitic acid or oleic acid treatment HSC-T6cells.1、200mmol/LResults:1.Palmitic acid and oleic acid induced HSC-T6re-lipidation by increasing theformation of lipid drops; the amount of lipid deposition drops in oleic acidintervention group significantly higher than control group,(P<0.05); the amount oflipid deposition drops in palmitic acid intervention group significant higher thancontrol group (P<0.05).2.The transfection efficiency of COX-2shRNA more than70%, meet the experimentrequirements.3. The expression of COX-2was increased in200mmol/L palmitic acid or20μg/mloleic acid treatment HSC-T6; The oleic acid intervention group was significantly higher than that control group (P<0.05); The palmitic acid intervent group wassignificantly higher than that control group (P<0.05). Compared With palmitic acidor oleic acid treatment group, COX-2shRNA+palmitic acid and COX-2shRNA+oleic acid treatment group significantly reduced COX-2expression, with a significantdifference (P<0.05).4.200mmol/L palmitic acid or20μg/ml oleic acid could increase the expression ofAcsl1, Acsl5, Acsl6,with a significant difference (P<0.05), while have no effect on theexpression of Acsl3, Acsl4(P>0.05); COX-2shRNA+palmitic acid and COX-2shRNA+oleic acid group further up-regulates the expression of Acsl5and Acsl6(P<0.05), but have no effect on the increase of Acsl1、Acsl3、Acsl4(P>0.05).Conclusion：1.Palmitic acid or oleic acid could induce the re-lipidation of HSC-T6.2.Palmitic acid or oleic acid up-regulated the expression of Acsl1、Acsl5、Acsl6inrat HSC-T6.3.COX-2inhibited the expression of Acsl5、Acsl6up-regulated by Palmitic acid oroleic acid.