Promoting Effect Of High Concentration Chlorogenic Acid On Lipid Accumulation In Hepatic Cells

Objective:The aim of this research was to investigate the effect of chlorogenic acid(CGA)on lipid accumulation and lipid peroxidation in human normal hepatic cells line L02.Methods:L02 cells were treated with TO901317(5?M),which is liver X receptor a(LXRa)agonist,and the intermixture of oleic acid(OA)-palmitic acid(PA)(666?M:333?M)to establish the pathological model of hepatic lipid accumulation and lipid steatosis in vitro.L02 cells were exposed to different concentrations of CGA for 24h.Using MTT test to detect the cell viability,Oil red O staining to check the lipid accumulation in hepatic cells,reactive oxygen species(ROS)assay kit to test the content of ROS,real-time polymerase chain reaction(RT-PCR)and western Blot to investigate the expression level of mRNA and protein of sterol regulatory element binding protein-lc(SREBP-1C),satatin-like phospholipase domain-containing protein 3(PNPLA3),which associated with hepatic cell's lipid accumulation,and the expression level of mRNA and protein of cytochrome P450 2E1(CYP 2E1),which closely related to lipid peroxidation.Results:1.The effect of CGA on L02 cell viability(1)After L02 cells were treated with CGA for 24h,when CGA concentration was lower than 20?M,it have no obvious effect on L02 cell viability;when CGA concentration was at 20?2000p?M,it promoted L02 cell viability significantly;when CGA concentrations was higher than 3mM,it inhibited L02 cell viability.(2)After combine CGA and TO901317 to treat cell for 24h,when CGA concentration was lower than 2mM,it promoted L02 cell viability significantly;when CGA concentrations was higher than 3mM,it inhibited L02 cell viability significantly.(3)After combine CGA and OA-PA to treat L02 cells for 24h,when CGA concentration was lower than 2mM,it promoted L02 cells significantly;when CGA concentrations was higher than 3mM,it inhibited L02 cell viability significantly.2.Effect of CGA on lipid accumulation and lipid peroxidation in L02 cells After CGA(0.5,1,2mM)treated L02 cells for 24h.CGA induced lipid accumulation,enhanced the content of ROS,and up-regulated the level of mRNA and protein of CYP 2E1,SREBP-1C,PNPLA3 in L02 cells.(2)After combine TO901317(5?M)and CGA(0.5,1,2mM)to treat L02 cells for 24h.TO901317 induce L02 cells lipid accumulation,enhanced the content of ROS,and up-regulated the level of mRNA and protein of CYP 2E1,SREBP-1C,PNPLA3 in L02 cells.CGA further enhanced L02 cells lipid accumulation,the content of ROS,and the expression level of mRNA and protein of CYP 2E1,SREBP-1C,PNPLA3.(3)After combine OA-PA(666?M:333?M)and CGA(0.5,1,2mM)to treat L02 cells for 24h.OA-PA induced lipid accumulation,enhanced the content of ROS,and up-regulated the expression level of mRNA and protein of CYP 2E1,SREBP-1C,PNPLA3 in L02 cells.CGA further enhanced L02 cells lipid accumulation,the content of ROS and the expression level of mRNA and protein of CYP 2E1,SREBP-1C,PNPLA3.Conclusions:This research indicated that high concentration CGA promoted L02 cells lipid accumulation and lipid steatosis via up-regulated the expression of SREBP-1C and PNPLA3 under normal or lipid accumulation condition.High concentration CGA lead to lipid peroxidation via up-regulated the expression of CYP 2E1 and enhanced ROS content under normal or lipid accumulation condition.