我们的网站为什么显示成这样？

可能因为您的浏览器不支持样式，您可以更新您的浏览器到最新版本，以获取对此功能的支持，访问下面的网站，获取关于浏览器的信息：

|本期目录/Table of Contents|

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

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2009年第3期
页码:
313-316
栏目:
基础研究
出版日期:
2009-05-15

文章信息/Info

Title:
DIDS Attenuates ischemia/reperfusion injury-induced cardiomyocytes apoptosis through PI3-K/Akt signaling pathway
作者:
张卫卫; 刘艳; 师堂旺; 刘佳妮; 刘艳霞; 王晓明
第四军医大学西京医院老年病科，陕西 西安 710032
Author(s):
ZHANG Wei-wei;  LIU Yan;  SHI Tang-wang;  LIU Jia-ni;  LIU Yan -xia;  WANG Xiao-ming
DepartmentofGeriatrics,Xijing Hospital,FourthMilitaryMedicalUniversity,Xi’an710032,Shaanxi,China
关键词:
心肌细胞; 缺血/再灌注; 凋亡; PI3-K/Akt信号通路
Keywords:
cardiomyocyte;  ischemia/reperfusion;  apoptosis;  PI3-kinase/Akt
分类号:
R542.2
DOI:
-
文献标识码:
A
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-ONETM Homogeneous 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。
Abstract:
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 cardiomyocytes 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 MTT colorimetric, Hoechest-33258 staining and caspase (Apo-ONETM Homogeneous Caspase)-3 kit and Western blot. RESULTS ①DIDS 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); ②Pretreatment with LY294002, the increase of myocardial cells’ viability, decrease of apoptotic bodies and the increase of caspase-3 activity were significant blocked (P<0.01); ③There 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.

参考文献/References

[1］ Kajstura J, Cheng W, Reiss K, et al. Apoptosis and necrotic myocyte cell deaths are independent contributing variables of infarct size in rats［J］. Lab Invest, 1996, 74(1):86-107.

［2］ Gottlieb RA, Burleson KO, Kloner RA, et al. Reperfusion injury induces apoptosis in rabbit cardiomyocytes［J］. J Clin Invest, 1994, 94(4):1621-1628.

［3］ d’Anglemont de Taocssigny A, Souktani R, Henry P, et al. Volume-sensitive chloride channels(lCl,vol) mediate doxorubicin-induced apoptosis through apoptotic volume decrease in cadiomyocytes［J］. Fundam Clin Pharmacol, 2004, 18(5):531-538.

［4］ Wang XM, Takahashi N, Uramoto H, et al. Chloride channel inhibition prevents ROS dependent apoptosis induced by ischemia-reperfusion in mouse cardiomyocytes［J］. Cell Physiol Biochem, 2005, 16(4-6):147-154.

［5］ 王晓明，臧益民，龚卫琴. 钾、氯离子通道阻断剂对sturosporine诱导心肌细胞凋亡的调节作用的研究［J］. 第四军医大学学报, 2004, 25(9):783-787.

［6］ Liu AH, Cao YN, Liu HT, et al. DIDS attenuates staurosporine-induced cardiomyocyte apoptosis by PI3K/Akt signaling pathway: activation of eNOS/NO and inhibition of Bax translocation［J］. Cell Physiol Biochem, 2008, 22(1-4):177-186.

［7］ Fujio Y, Nguyen T, Weneker D, et al. Akt promotes survival of cardiomyocytes in vitro and protects against ischemia-reperfusion injury in mouse heart［J］. Circulation, 2000, 101(6):660-667.

［8］ Zhang KR, Liu HT, Zhang HF, et al. Long-term aerobic exercise protects the heart against ischemia/reperfusion injury via PI3 kinase-dependent and Akt-mediated mechanism［J］. Apoptosis, 2007, 12(9):1579-1588.

［9］ 刘海涛，张海峰，司瑞，等. 胰岛素保护缺血/再灌注心脏: PI3-K/Akt和JNKs信号通路间的交互作用［J］. 生理学报, 2007, 59(5):651-659.

［10］Uramoto H, Takahashi N, Dutta AK, et al. Ischemia-induced enhancement of CFTR expression on the plasma membrane in neonatal rat ventricular myocytes［J］. Jpn J Physiol, 2003, 53(5):357-365.

［11］曹亚南，刘安恒，张卫卫，等. 模拟缺血/再灌注诱导小鼠心肌细胞凋亡模型的构建及其生化特征［J］. 心脏杂志, 2008, 20(3):259-263.

［12］Cook SA, Poole-Wilson PA. Cardiac myocyte apoptosis［J］. Eur Heart J, 1999, 20(22):1619-1629.

［13］Araki T, Hayashi M, Watanabe N, et al. Down-regulation of Mcl-1 by inhibition of the PI3-K/Akt pathway is required for cell shrinkage-dependent cell death［J］. Biochem Biophys Res Commun, 2002, 290(4):1275-1281.

［14］d’Anglemont de Tassigny A, Berdeaux A, Souktani R, et al. The volume-sensitive chloride channel inhibitors prevent both contractile dysfunction and apoptosis induced by doxorubicin through PI3kinase, Akt and Erk 1/2［J］. Eur J Heart Fail, 2008, 10(1):39-46.

备注/Memo

备注/Memo:
收稿日期：2008-10-21.基金项目：国家自然科学基金（30570758，30770847） 通讯作者：王晓明，副教授，博士，主要从事心血管疾病的基础与临床研究Email: xmwang@fmmu.edu.cn 作者简介：张卫卫，硕士生Email:weiweizhang1982@hotmail.com
更新日期/Last Update: 2009-05-18