COX-2Controls The HSC-T6Cell Proliferation And ACSL Family Genes Expression By Arachidonic Acid

ObjectiveThis study aims at clarifying the mechanism of COX-2controlling the HSC-T6cell proliferation and ACSL family gene expression in terms of arachidonic acid.Methods11. Use the MTT assay to test the growth of rat’s HSC-T6cells when they are in AA20μM,40μM,60μM.2. Use RT-PCR method to detect the expression of ASCL1, ACSL3, ACSL4,ACSL5, ACSL6,COX-2in HSC by different concentrations of AA.Results11. Arachidonic acid40μM can remarkablely promote the proliferation of HSC.3. The research demonstrates that the expression level of ACSL6and ACSL3inAA40μM group and AA60μM group is remarkably higher than the HSC group(p<0.05). The level of COX-2in AA20μM group and AA40μM group raises thanHSC group (p<0.05).The expression level of ACSL1in AA60μM is relative grows.The expression of ACSL4and ACSL5has no significant changes in each group(p>0.05).Methods21. Transfer the constructed pYr-1.1-hU6-EGFP-COX-2shRNA1into HSC, assessthe transfection efficiency under the fluorescence microscope.Transfection groups:HSC group,AA40μM group,empty vector transfection+AA group,COX-2shRNA+AA group.Choose the optim40μM AA concentration.2. Use Western blot method to detect the expression of COX-2, ACSL family gene after transfer48h.Results21. AA40μM can promote the proliferation of HSC which compareed with HSCgroup.COX-2shRNA1+AA group decreased the proliferation of HSC comparewith pYr-1.1HK+AA group and had not significantly elevated in the proliferationin contrast to control group.2. The expression of COX-2,ACSL1,ACSL6in AA group increased significantly thanthe control group (p<0.05).COX-2shRNA1+AA group decreased COX-2,ACSL1,ACSL6expression of HSC compare with pYr-1.1HK+AA group.There wasno change between COX-2shRNA1transfected group and the control group.Conclusions1. COX-2can stimulate growth of HSC by the way of AA.2. COX-2increase HSC’s ACSL1,ACSL6gene expression through arachidonic acid.