Caspase-3抑制剂对兔体外循环心肌细胞凋亡的影响

Effect of Caspase-3 inhibitor on cardiocyte apoptosis after cardiopulmonary bypass in rabbits

目的:探讨Caspase-3抑制剂对兔体外循环心肌细胞凋亡的作用。方法:将24只健康新西兰大耳白兔随机分为3组:正常对照组、体外循环(cardiopulmonary bypass,CPB)组和Caspase-3抑制剂组。采用原位末端标记(TUNEL)法观察心肌细胞凋亡的改变;借助Caspase-3活性检测试剂盒,荧光比色法检测心肌细胞凋亡过程中Caspase-3活性的变化;应用半定量RT-PCR技术分析心肌细胞凋亡过程中Caspase-3 mRNA的表达。结果:Caspase-3抑制剂组心肌细胞凋亡指数(apoptosis index,AI)为(4.24±1.36)%,与CPB组(7.28±2.28)%相比明显降低(P<0.01);Caspase-3抑制剂组的Caspase-3活性比CPB组降低了47.44%(P<0.01);Caspase-3 mRNA的表达分析显示,Caspase-3抑制剂组明显低于CPB组(P<0.01)。结论:Caspase-3抑制剂对兔体外循环心肌细胞凋亡具有明显的抑制作用,其机制与抑制凋亡途径中关键蛋白酶Caspase-3的活性有关。

Objective： To observe the effect of Casepase-3 inhibitor on cardiocyte apoptosis after cardiopulmonary bypass in rabbits. Methods： Twenty-four healthy rabbits were randomly divided into 3 groups; control group, CPB group and Casepase-3 inhibitor group. Cardiocyte apoptosis was detected by TUNEL; the Casepase-3 activity was detected by Casepase-3 activity assay kit. Applying semiquantu RT- PCR to detect the expression of Caspase-3 mRNA. Results： TUNEL method showed that cardiocyte apoptosis indexes in Casepase-3 inhibitor group was （4.24 ± 1.36 ）%, and compared with CPB group （7.28 ± 2.28） %, was significantly lower （ P 〈 0. 01 ）. Casepase-3 activity in Casepaseo3 inhibitor group was 47.44% less than CPB group ; expression of Caspase-3 mRNA analysis showed that expression of Caspase-3 mRNA in Casepase-3 inhibitor group was very lower than that in CPB group （P 〈 0.01 ）. Conclusion： Casepase-3 inhibitor efficiently inhibited cardiocyte apoptosis after cardiopulmonary bypass in rabbits, and inhibited activity of critical protease Caspase-3 in apoptosis.