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目的观察丝裂原活化蛋白激酶(MAPK)抑制剂PD98059对氯化镉(CdCl2)诱导大鼠肾上皮细胞(NRK)凋亡基因Bcl-2和Caspase-3 mRNA表达的影响。方法应用流式细胞仪测定CdCl2染毒6 h的NRK细胞凋亡作用;用MAPK抑制剂PD98059预先处理NRK细胞1 h后,用不同浓度镉染毒,运用实时荧光定量PCR技术,检测NRK肾细胞内凋亡基因Bcl-2和Caspase-3 mRNA表达水平。结果NRK细胞的凋亡率与CdCl2染毒存在剂量-效应关系;2.5,5,10μmol/L镉染毒使Bcl-2 mRNA的表达水平分别下降至对照组的72%,53%和37%,而Caspase-3 mRNA表达水平上升为对照组的125%,153%和175%。在细胞培养基中预先加入PD98059然后用镉染毒,PD98059可明显阻止镉引起的Bcl-2 mRNA表达水平下降以及Caspase-3 mRNA表达水平升高。结论MAPK抑制剂PD98059对镉引起凋亡基因Bcl-2和Caspase-3的表达水平改变具有明显干预作用,提示MAPK信号调节可能参与镉诱导的细胞凋亡过程。
Objective To investigate the effects of mitogen-activated protein kinase (MAPK) inhibitor PD98059 on the expression of Bcl-2 and Caspase-3 mRNA induced by cadmium chloride (CdCl2) in rat renal epithelial cells. Methods Flow cytometry was used to determine the apoptosis of NRK cells exposed to cadmium chloride (CdCl2) for 6 h. NRK cells were pretreated with MAPK inhibitor PD98059 for 1 h and then exposed to different concentrations of cadmium. Real-time fluorescent quantitative PCR was used to detect NRK cells The expression of Bcl-2 and Caspase-3 mRNAs were detected by Western blot. Results The apoptotic rates of NRK cells were dose-dependently correlated with CdCl2 exposure. The levels of Bcl-2 mRNA were decreased to 72%, 53% and 37% of the control cells exposed to 2.5, 5 and 10μmol / The Caspase-3 mRNA expression level increased to 125%, 153% and 175% of the control group. PD98059 was pre-added to the cell culture medium and then treated with cadmium. PD98059 significantly prevented the decrease of cadmium-induced Bcl-2 mRNA expression and the increase of Caspase-3 mRNA expression. Conclusions The MAPK inhibitor PD98059 can significantly influence the expression of Bcl-2 and Caspase-3 induced by cadmium, suggesting that MAPK signaling may be involved in the process of cadmium-induced apoptosis.