Effect Of ShRNA Targeting COX-2 On The Gene Expression Of Cox-2, Leptin In Hepatic Stellate Cell

Objective:Research the gene expression of COX-2, Leptin in HSC which were transfacted by COX-2shRNA1,COX-2shRNA2,COX-2shRNA3.Methods:1.Transfering COX-2shRNA vector into HCS by Lipofectamine2000,after 24h,48h and 72h, fluorescence microscopy to assess cell's transfection efficiency of plasmid.2.RT-RCR measure the mRNA expression of COX-2, and leptin in HSC which were inhibited by COX-2shRNA1,COX-2shRNA2,COX-2shRNA3 after 24h,48h and 72h.3.Western measure the protein expression of COX-2 in HSC which were inhibited by COX-2shRNA1,COX-2shRNA2,COX-2shRNA3 after 48h.4.Immunocytochemistry measure the protein expression of COX-2, leptin in HSC which were inhibited by COX-2shRNA1.Results:1.Transfering COX-2shRNA vector into HCS by Lipofectamine2000,after 24h,48h and 72h, three sequences of the three time periods of transferred HSC were expressed fluorescence2. RT-PCR demonstrated the expression of COX-2, leptin can be restrained by COX-2shRNA1,COX-2shRNA2,COX-2shRNA3 ,the best inhibited effect is COX-2shRNA1 after 72h.3.Western demonstrated the protein expression of COX-2 were inhibited by COX-2shRNA1,COX-2shRNA2,COX-2shRNA3, and the best inhibited effect is COX-2shRNA1 after 48h. 4.Immunocytochemistry demonstrated the expression of COX-2,leptin in HSC can be inhibited by COX-2shRNA1.Conclusions:1.COX-2 expression in HSC should be inhibited Signifieantly by three targeting vector of COX-2shRNA,and the best inhibited effect is COX-2shRNA1;2. Leptin expression in HSC should be inhibited Signifieantly by inhibiting gene expression of COX-2 .