Nimesulide May Promote The Apoptosis Of HSC-T6 By Caspase-3

Objective: To prove the effect of nimesulide(the selective COX-2 inhibitor) on the expression of Caspase-3 protein,in order to provide a new idea for further clarifying the mechanism of COX-2 inhibiting HSC apoptosis and promoting liver fibrosis. Methods: 1. Cell culture: before starting experiment,cultured a rat hepatic stellate cell line(HSC-T6) with RPMI-1640 medium supplemented with 10% Biological industries in incubator. Incubation condition were at 37℃ under 5%CO2 at constant humidity.Changed the culture medium every 2-3day. 2. the groups and methods of the experiment: this experiment were divided into three groups:the blank control group, 200umol/L NS treatment group(NS200 group for short) and 400umol/L NS treatment group-(NS400 group for short).After treated HSC-T6 with different concentrations of NS for 48 h, the apoptosis and cell cycle of HSC-T6 were detected using FCM assay. Meanwhile,the protein expression of Caspase-3 was estimated using WB analysis.Results: 1.The impact of NS on the apoptosis of HSC-T6: FCM assay showed that the apoptosis rate of the HSC-T6 in the blank control group,NS-200 group and NS400 group were 10.23%±1.34%, 25.23%±7.7%,65.87%±2.89%, respectively. There were significant difference in the apoptosis rate of the HSC-T6 cells among the three groups(P<0.05),and compared with the blank control group,the apoptosis rate of the HSC-T6 in the NS- 200 group and NS400 group increased significantly(P<0.05).It indicated that nimesulide could promote dose-dependently the apotosis of HSC-T6. 2.The impact of NS on the cell cycle of HSC-T6: cell cycle analysis showed cell viability was almost unaffected by treatment with NS(200umol/l) for 48 h. However, there were a growth retardation in G0/G1 phase of cell cycle of the HSC-T6 after treatment with NS( 400 umol/l) 48 h. It incated that nimesulide could arrest dose-dependently the cell cycle of HSC-T6. 3.The impact of NS on the protein expression of Caspase-3:WB analysis showed that compared with the blank control group, the protein expression of Caspase-3 in the NS 200 group and NS 400 group were increased obviously and dose-dependmently.Conclusion: 1. NS may promote dose-dependmently the apoptosis of HSC-T6. 2. NS may arrest dose-dependmently the cell cycle progression of HSC-T6. 3. NS may promote the apoptosis of HSC-T6 and arrest the cell cycle of HSC-T6 by upgraduating the protein expression of Caspase-3.