Effect Of COX-2shRNA On Expression Of Atf3、Serpinb2、 Pgk1、Thra In Rat Hepatic Stellate Cells

ObjectiveThis study research the Gene expression of Atf3, Serpinb2, Pgk1and Thra inHepatic Stellate Cell(HSC-T6) by the way of COX-2shRNA.MethodsThis experiment is divided into three groups: the control group (PBS alternativeplasmid), empty vector group (DNA fragments of not inserted the target gene, HKplasmid) and the transfection group (COX-2shRNA1group). The plasmid transfectedinto rat hepatic stellate cells by LipofectamineTM2000, observe its transfectionefficiency by fluorescence microscopy, After24h,48h and72h by transfer COX-2shRNA,extract cell RNA, detect the expression of Atf3、Serpinb2、Pgk1、Thraby the way of RT-PCR technology.Results1.immunocytochemistry showed: After24h,48h and72h by transfer COX-2shRNA1,we can visible green fluorescent in the cytoplasm of the cells under a fluorescencemicroscope,the transfection efficiency can be up to70%above, The blank group andempty vector have not green fluorescence in cytoplasmic.2.The expression of COX-2: After24h,48h and72h by transfer COX-2shRNA,COX-2mRNA expression was lower than the control group and empty vector of bythe way of RT-PCR（P<0.05, The72h’inhibition was especially noticeable, and theblank control group and empty vector group were without any silence effect.3.The expression of Atf3、Serpinb2and Thra: After24h,48h and72h by transferCOX-2shRNA, Atf3and Serpinb2mRNA expression was lower than the controlgroupand empty vector of by the way of RT-PCR（P<0.05）,The72h’inhibition wasespecially noticeable, and the blank control group and empty vector group were without any silence effect.4.The expression of Pgk1：24later than the transfection, no difference wasobserved, under the help of RT-PCR technology, in expression of Pgk1mRNA amongtransfection group, blank control group and empty vector group（P<0.05）. Howeverexpression of Pgk1mRNA in transfection group was lower than that of blank controlgroup and empty vector group at the time of48h and72h. The72h’inhibition wasespecially noticeable,and the blank control group and empty vector group werewithout any silence effect.Conclusions1.The transfection of targeted COX-2shRNA into rats HSC-T6can result in silent ofexpression of COX-2.2.Targeted COX-2shRNA can decrease the expression of Serpinb2in HSC,genes related to proliferation.3.Targeted COX-2shRNA can reduce the expression of Pgk1、Thra in HSC, genesrelated to lipid metabolism.4.Targeting COX-2shRNA inhibit of the the proliferation of HSC may be throughadjusting the expression of the Serpinb2, Pgk1, Thra and other gene.