氯通道阻滞剂对大鼠心肌缺血再灌注损伤致心肌细胞凋亡的作用  

The effects of chloride ion channel inhibition on myocardial apoptosis induced by ischemia-reperfusion injury

作　　者：陈涛 雷兰萍 周和平 金振晓 

机构地区：[1]第四军医大学第一附属医院心血管外科,西安710032

出　　处：《中国体外循环杂志》2011年第2期85-88,共4页Chinese Journal of Extracorporeal Circulation

基　　金：陕西省自然科学基金资助项目(2007C224);西京医院学科助推计划项目(XJZT09M16)

摘　　要：目的本研究应用大鼠在体心肌缺血再灌注损伤(I/RI)动物模型,观察氯通道阻滞剂DIDS(4,4'-disothiocya-nostibibene-2,2'-disulfonic acid)对心肌细胞凋亡的作用以及与磷脂酰肌醇3激酶蛋白激酶B(PI3K-Akt)信号通路可能存在的相互作用,明确DIDS心肌保护作用的可能机制。方法 75只大鼠随机分成5组:假手术组,I/RI组,I/RI+DIDS(14 mg/kg),I/RI+LY294002(PI3K-Akt特异性阻断剂),I/R+DIDS+LY294002组。TTC染色检测心肌梗死范围;TUNEL法测定细胞凋亡;Caspase-3试剂盒检测caspase-3活性;免疫印迹法检测p-Akt水平。结果与I/R组比较DIDS可以减少心肌梗死范围(各组n=4,P<0.01),可以明显抑制心肌细胞凋亡(各组n=5,P<0.01),降低凋亡效应子caspase-3活性(各组n=5,P<0.01),提高I/RI心肌的p-Akt水平(各组n=5,P<0.01),LY294002可以部分阻断这些作用,LY294002自身并无减少心肌梗死范围的作用。结论氯离子通道阻滞剂DIDS对缺血再灌注损伤的心肌具有保护作用,其作用部分是通过PI3K/Akt信号通路来实现的。OBJECTIVE To investigate the effects of chloride channel blocker 4,4＇-disothiocyanostibibene-2,2＇-disulfonic acid（DIDS） on myocardium underwent ischemia and reperfusion injury（I/RI）.METHODS Seventy-five male healthy SD rats were randomized into five groups（n=15）： group A,sham operation group;group B,I/RI group,ischemia 30 min,reperfusion 4 h;group C,I/RI＋DIDS group： ischemia 30 min,DIDS was intravenously administered at 7 mg/kg/h exactly at the starting of reperfusion for 2 h;group D,I/RI＋LY294002 group： ischemia 30 min,LY294002（0.3 mg/kg） was infused during the second 15 min of ischemia;group E,I/R＋DIDS＋LY294002 group： rats were treated with combination of C and D group treatment procedure.After 4 h reperfusion,hearts were harvested.Myocardial infarct size,apoptotic index of cardiomyocytes,caspase-3 activity and p-Akt levels were detected.RESULTS DIDS reduced myocardial infarct size（n=4,P0.01 vs.I/RI group）,inhibited cardiomyocytes apoptosis（n= 5,P0.01 vs.I/RI group）,decreased caspase-3 activity（n=5,P0.01 vs.I/RI group） and significantly increased p-Akt levels（n= 5,P0.01 vs.I/RI group）;LY294002 partially blocked these effects except the levels of p-Akt.But LY294002 itself had no effect on myocardial infarct size.CONCLUSION DIDS play a protective role on myocardium underwent ischemia and reperfusion injury,and its protective effect is partially through PI3K/Akt signal pathway.

关 键 词：缺血再灌注损伤 氯离子通道 凋亡 PI3K/AKT 

分 类 号：R965[医药卫生—药理学]