论文部分内容阅读
目的探讨硫化氢(H2S)在p38MAPK信号通路对大鼠肝星状细胞(hepatic stellate cell,HSC)凋亡中的作用及磷酸化P38、Caspase-3蛋白表达的变化。方法实验设对照组(HSC加含10%胎牛血清的DMEM培养液)、二甲基亚砜(DMSO)组(对照组基础上加DMSO,使其终浓度为0.1%)、NaHS组(对照组基础上加NaHS,使其终浓度为50μmol/L)、SB组(DMSO组基础上加SB203580,使其终浓度为75μmol/L)、SB加NaHS(SB+NaHS)组;采用Hoechst荧光染色检测细胞凋亡;Western blotting法检测磷酸化p38MAPK表达及Caspase-3蛋白表达水平。结果与对照组比较,SB组和SB+NaHS组HSC-T6的凋亡率增加(P<0.05),NaHS组p38MAPK磷酸化水平及Caspase-3表达均明显增高(P<0.01);与NaHS组比较,SB组和SB+NaHS组细胞凋亡率增加明显(P<0.01),p38MAPK磷酸化水平表达降低(P<0.01);SB+NaHS组较SB组Caspase-3蛋白表达升高(P<0.05)。结论 p38MAPK及Caspase-3在H2S刺激的HSC-T6中表达增强,H2S能促使SB203580诱导的HSC-T6细胞凋亡,其作用机制可能与活化p38MAPK的磷酸化途径,进而激活Caspase-3的表达有关。
Objective To investigate the effect of hydrogen sulfide (H2S) on the apoptosis of rat hepatic stellate cells (HSC) induced by p38 MAPK and the expression of phosphorylated P38 and Caspase-3 proteins. Methods The control group (HSC plus 10% fetal bovine serum in DMEM), dimethyl sulfoxide (DMSO) group (DMSO added to the control group at a final concentration of 0.1%), NaHS group (SBHS) plus NaHS (final concentration was 50μmol / L), SB group (SB203580 supplemented with DMSO at the final concentration of 75μmol / L), SB plus NaHS group (SB + NaHS) Apoptosis was detected. The phosphorylation of p38MAPK and the expression of Caspase-3 protein were detected by Western blotting. Results Compared with the control group, the apoptosis rate of HSC-T6 in SB group and SB + NaHS group was increased (P <0.05), and the phosphorylation of p38MAPK and the expression of Caspase-3 in NaHS group were significantly increased (P <0.01) Compared with SB group and SB + NaHS group, the apoptosis rate increased significantly (P <0.01) and the phosphorylation level of p38 MAPK decreased (P <0.01). The expression of Caspase-3 protein increased in SB + NaHS group compared with SB group (P < 0.05). Conclusions The expression of p38MAPK and Caspase-3 in H2S-stimulated HSC-T6 cells is enhanced, and H2S can promote the apoptosis of HSC-T6 cells induced by SB203580, which may be related to the activation of p38MAPK phosphorylation pathway and activation of Caspase-3 expression .