He Aina, Wang Jian-an, Gui Chun, Jiang Yun, Sun Yong, Chen Tielong
Heart Center, Second Affiliated Hospital, College of Medicine, Zhejiang University, 88 Jiefang Road, Hangzhou 310009, China.
Folia Histochem Cytobiol. 2007;45(4):397-400.
The role of mitochondrial apoptotic pathway in cardiomyocytes subjected to hypoxia/reoxygenation(H/R) was studied. Cultured cardiomyocytes from neonatal Sprague-Dawley rats were exposed to hyoxia/reoxygenation, the apoptotic cardiomyocytes were stained with Annexin-V-FITC, Hoechst 33342 and TUNEL assay. Mitochondrial transmembrane potential of cardiomyocytes was assessed by JC-1 under fluorescence microscope, the expressions of bcl-2, bax, cytochrome c, apoptosis-induced factor (AIF), and caspase-3 were tested by western-blot. Our data showed apoptosis of cardiomyocytes was significantly increased during H/R, accompanied by translocation of bax to mitochondria, release of cytochrome c and AIF to cytosol. The results indicate that the mitochondrial-mediated apoptotic pathway is initiated as a result of H/R.
研究了线粒体凋亡途径在经历缺氧/复氧(H/R)的心肌细胞中的作用。将新生Sprague-Dawley大鼠培养的心肌细胞暴露于缺氧/复氧环境,用膜联蛋白V-异硫氰酸荧光素(Annexin-V-FITC)、Hoechst 33342染色以及TUNEL法检测凋亡心肌细胞。在荧光显微镜下用JC-1评估心肌细胞的线粒体跨膜电位,通过蛋白质免疫印迹法检测bcl-2、bax、细胞色素c、凋亡诱导因子(AIF)和半胱天冬酶-3的表达。我们的数据显示,在缺氧/复氧期间心肌细胞凋亡显著增加,同时伴有bax转位至线粒体、细胞色素c和AIF释放至胞质溶胶。结果表明,缺氧/复氧引发了线粒体介导的凋亡途径。