Caspase-3在心衰兔模型细胞凋亡过程中的变化  

Changes of Caspase-3 Expression during Myocyte Apoptosis in the Rabbit Model of Congestive Heart Failure

作　　者：王萧[1] 林传权[1] 许庆文[2] 付云[1] 谢玲玲[1] 刘金元[3] 

机构地区：[1]广州中医药大学实验动物中心,广州510405 [2]广州中医药大学临床药理所,广州510405 [3]广州中医药大学测试中心,广州510405

出　　处：《中国实验动物学报》2009年第3期201-205,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：广东省科技计划资助项目(编号:2005B60302006)

摘　　要：目的研究caspase-3在导管术致超容量负荷型心衰兔模型细胞凋亡中的变化。方法运用导管术造成兔的超容量负荷型心衰模型,然后分别于造模后24 h和造模后7 d处死兔,取心脏组织做电镜检查;通过流式细胞仪检测心肌细胞凋亡情况、caspase-3试剂盒检测其活性、Western blot法检测caspase-3蛋白水平的表达。结果①电镜检查表明:正常组心内膜完整无损伤,有少量红细胞附着;造模24 h后和造模7 d后均出现大量红细胞附着和血栓,且造模7 d的血栓较造模24 h减少。②AnnexinV联合PI染色结果表明:正常组凋亡率较低(3.72±1.92)%,随着时间推移,凋亡细胞所占比例呈上升趋势,造模7 d(10.73±2.27)%明显高于造模24 h(7.62±1.70)%。③Caspase-3活力测定:随时间推移,caspase-3活力呈上升趋势,造模7 d明显高于造模24 h,指数分别为2.7±0.31和2.15±0.32。④Western Blot检测结果表明:正常组基本检测不到激活的caspase-3蛋白酶,在造模后24h和造模后7 d caspase-3蛋白酶均有明显表达,但造模7 d有所下降。结论超容量负荷型心衰兔模型的细胞凋亡与caspase-3有密切相关。Objective To observe the changes of caspase-3 in the rabbit model of congestive heart failure with volume overload caused by catheterization. Method The model of congestive heart failure with volume overload was set up by catheterization. The rabbits were sacrificed and the hearts were taken out for scanning electron microscopic examination at 24 h and 7 d after modeling, respectively. The apoptosis in myocardial cells was detected by flow cytometry. The activity of caspase-3 was assessed by caspase-3 assay kit. At the same time, the expression of caspase-3 protease was determined by Western blot analysis. Results ①The electron microscopic examination showed few erythrocytes adhered on the endothelial surface without any other alterations on the endocardium in normal rabbits, but many red ceils and thrombi were observed on the endocardium in the 24 h and 7 d experimental groups. The areas of thrombi in the 7 d group were smaller than those in the 24 h group. ②The staining of annexin-V combined with PI showed： The rate of apoptotic ceils was going up as time went on, the ratio in the normal group, 24 h and 7 d model groups were （3.72± 1.92, % ）, （ 10.73 ± 2.27% ） and （7.62 ±1.70% ）, respectively. ③The activity of caspase-3 was increasing, from 2.15 ± 0.32 （24 h） to 2.7 ± 0.31 （7 d）. O The results of Western blot analysis showed no expression of the active form of caspase-3 in the nomal group, but significantly increased in the 24 h group and 7 d model groups, and the level of 7 d group became decreased. Conclusion Apoptosis of myocardial cells has a very close relationship with caspase-3 protease in the rabbit models of congestive heart failure with volume overload.

关 键 词：心肌组织 心衰模型 新西兰兔 细胞凋亡 CASPASE-3 

分 类 号：R542.2[医药卫生—心血管疾病]