Inhibitory Effects Of Long Chain Acyl Coenzyme A Synthetase On The Proliferation Of HSC-T6 By COX-2

Objective: To study the effect of inhibition of long chain acyl coenzyme A synthetase activity on the proliferation of HSC-T6 regulated by COX-2Methods:1?HSCs-T6 cells were cultured in vitro,the constructed p Yr-1.1COXsh RNA was transfected into activated HSCs-T6 cells,the transfection efficiency was observed under fluorescent microscope.a)vector group(NC group): HSCs-T6 cells activated by +p Yr-1.1-HK;b)COX-2sh RNA group(KD group): activated HSCs-T6+ p Yr-1.1-COX2 sh RNA cells;c)NC+Acsl inhibitor group: activated HSCs-T6 cells +p Yr-1.1-HK+ Acsl inhibitor Triacsin C;d)KD+Acsl inhibitor group: HSCs-T6 cells activated+p Yr-1.1-COX-2sh RNA+Acsl inhibitor Triacsin C.2?MTT method was used to detect the proliferation of transfected HSCs-T6 cells.3?RT-PCR technique was used to detect the expression of a-SMA,COX-2,Acsl1,Acsl3,Acsl4,Acsl5,Acsl6 gene in HSCs-T6 cells of each group after transfection of 48 H.ResuLts:1?The green fluorescent plasmid was found in HSCs-T6 cells and the transfection rate was over 75%;2?With the blank control group,empty vector group compared with the COX-2sh RNA plasmid and 48 h HSCs-T6 cell proliferation rate decreased,and there was statistical significance(P < 0.05);the rate of 72 h significantly decreased the proliferation,and there was statistical significance(P < 0.05);3?a-SMA,COX-2,Q-PCR test results of the Acsl family: compared with the empty vector group,COX-2sh RNA group,a-SMA,COX-2 Acsl6 expression decreased,m RNA expression increased significantly,no significant other subtypes of Acsl expression;compared with COX-2sh RNA group,COX-2sh RNA inhibitor of +Acsl group a-SMA,COX-2 expression increased,Acsl6 decreased expression of m RNA.4.MTT to detect cell viability results: compared with NC group and KD group,NC+ inhibitor of Acsl group promoted the proliferation of HSCs-T6 cells;compared with KD group,KD+ inhibitor of Acsl group promoted the proliferation of HSCs-T6 cells;compared with NC+Acsl inhibitor group,KD+ inhibitor of Acsl group inhibited the proliferation of HSCs-T6 cells..Conclusions:1?COX-2 Sh RNA can effectively inhibit the proliferation of HSCs cells.2?Inhibition of Acsl activity can reverse the silencing of COX-2 on the proliferation of HSC-T6,suggesting that COX-2 may regulate HSC-T6 proliferation through Acsl pathway.