DIDS通过PI3-K/Akt途径减弱缺血/再灌注损伤诱导心肌细胞凋亡  

DIDS Attenuates ischemia/reperfusion injury-induced cardiomyocytes apoptosis through PI3-K/Akt signaling pathway

作　　者：张卫卫[1] 刘艳[1] 师堂旺[1] 刘佳妮[1] 刘艳霞[1] 王晓明[1] 

机构地区：[1]第四军医大学西京医院老年病科,陕西西安710032

出　　处：《心脏杂志》2009年第3期313-316,共4页Chinese Heart Journal

基　　金：国家自然科学基金(30570758;30770847)

摘　　要：目的探讨磷脂酰肌醇3激酶(PI3-K)/蛋白激酶B(Akt)信号通路在DIDS(4,4′-diisothiocyanostilbene-2,2′-disulfonic acid)减弱缺血/再灌注损伤(I/RI)诱导心肌细胞凋亡中的作用。方法以I/RI诱导心肌细胞凋亡,然后用PI3-K特异性的抑制剂LY294002[22(42吗啉基)282苯基24氢212苯并吡喃242酮]进行干预。实验分为正常对照组、I/RI组、I/RI+DIDS组和I/RI+DIDS+LY294002组。通过噻唑蓝(MTT)比色法、Hoechest-33258染色和半胱天冬蛋白酶(Apo-ONETMHomogeneous Caspase)-3试剂盒以及Western blot分别检测:心肌细胞的存活率(%)、细胞核的形态变化和caspase-3活性、Akt的磷酸化。结果①DIDS能够显著抑制I/RI诱导的细胞存活率的下降,抑制凋亡小体的出现,抑制caspase-3的活性的增加(P<0.01)。②用LY294002预处理后,DIDS保护I/RI心肌细胞存活的作用减弱、凋亡小体增加和caspase-3活性明显升高(P<0.01)。③I/RI组与正常对照组Akt磷酸化无明显差异,DIDS能够显著增加I/RI组中Akt蛋白的磷酸化(P<0.01)。用LY294002预处理后,DIDS对I/RI组Akt蛋白磷酸化的影响明显减弱(P<0.01)。结论DIDS可通过激活PI3-K/Akt信号通路减弱I/RI诱导的心肌细胞的凋亡。AIM To explore the role of phosphatidylinositol 3＇-kinase （PI3-K）/protein kinase B （Akt） signaling pathway during DIDS （4,4＇-diisothiocyanostilbene-2,2＇-disulfonic acid） inhibited cardiomyo- cytes apoptosis induced by ischemia/reperfusion injury （I/RI）. METHODS Cardiomyocytes apoptosis was induced by I/RI, then it was treated with phosphatidylinositol 3 kinas＇ inhibitor LY294002 [ 22 （42- Morpholine） 282 H-24-212 benzopyran 242] （specific inhibitor of PI3-K）. Experiment was divided into the normal control group, I/RI group, I/RI ＋ DIDS group and I/RI ＋ DIDS ＋ LY294002 group. The cell viability, morphology changes of nucleus and caspase-3 activity, Akt phosphorylation was observed through the MTY colorimetric, Hoechest-33258 staining and caspase （Apo-ONETM Homogeneous Caspase）-3 kit and Western blot. RESULTS （1） DIDS was able to markedly inhibit the I/RI indicated by a decline of cell viability, inhibited the emergence of apoptotic bodies, restrained the increase of caspase-3 activity （P 〈 0. 01 ） ; （2） Pretreatment with LY294002, the increase of myocardial cells＇ viabil- ity, decrease of apoptotic bodies and the increase of caspase-3 activity were significant blocked （P 〈 0.01 ） ; （3） There was no significant difference of Akt phosphorylation between I/RI group and control. DIDS could dramatically increase Akt phosphorylation in I/RI group （P 〈 0.01 ） ; Pretreatment with LY- 294002, DIDS＇ effect on the Akt phosphorylation in the I/RI group was decreased significantly （P 〈 0. 01）. CONCLUSION DIDS attenuates I/RI-induced myocardial apoptosis through the activation of PI3K/Akt signaling pathway.

关 键 词：心肌细胞 缺血/再灌注 凋亡 P13一K/Akt信号通路 

分 类 号：R542.2[医药卫生—心血管疾病]