Effects Of ShRNA Targeting COX-2 On Metabolism Of Triglyceride,Cholesterol,Vitamin A In Hepatic Stellate Cells(HSC-T6)

Object:The aim of this study was to investigate the over expression and co-expression of COX-2 and some important genes of lipid metabolism such as ACC,HMGCR,CPT1a in activated hepatic stellate cell, and to find their relationship to activated hepatic stellate cell.Cyelooxygenases-2(COX-2)-short interfering RNA is constructed so as to suppress the expression of COX-2 in activated hepatic stellate cell HSC-T6.We examined the effect of the down-regulation of COX-2 on expression of COX-2,ACC,HMGCR,CPT1a and on expression of triglyceride, cholesterol, vitamin-A, to find out the effects of COX-2 on hepatic fibrosis.Methods:Use passage HSC-T6 as study objects. The experiment is divided into 6 groups: The first group is shRNA 1 group; the second group is shRNA 2 group; the third group is shRNA 3 group;the forth group is blank group; the fifth group is Nimesulide group and the sixth group is non-sense shRNA group. And from group 3 to group 5, the cell is exposed to shRNA targeting COX-2 at the best concentration for 24h,48h,72h respectively, detect the expression of COX-2,ACC,HMGCR,CPT1a of he HSC-T6 of each group by RT- PCR respectively, and detect the intracellular amount of triglyceride , cholesterol ,vitamin-A through high performance liquid chromatography (HPLC),thereby make out the time– effect relation curve of effects of shRNA targeting COX-2 on lipid metabolism of hepatic stellate cells(HSC-T6).Using±s to represent experimental data, using SPSS13.0 statistical software to analyze the data, and P <0.05 is considered significantly important. Results:After exposing the cell to shRNA targeting COX-2 at the best concentration for 24h,48h,72h respectively,the result displays: COX-2 mRNA expression were obviously knocked down after transfection with shRNA, while the expression of HMGCR,CPT1 were decreased on the duration dependently,the expression of ACC were decreased on the duration dependently.The HPLC result says:the increase of TC and CE in the cells and decrease of cholesterol is dependent on the duration.Conclusions:1. COX-2 could incresde the expression of HMGCR,CPT1,and decrease the expression of ACC.2. When the COX-2 mRNA expression were obviously knocked down,the sum of lipid coml. In the lipid droplet of HSC, which could make the HSC retransformed.