L-3-n-butylphthalide protects rats' cardiomyocytes from ischaemia/reperfusion-induced apoptosis by affecting the mitochondrial apoptosis pathway
L-3-n-butylphthalide protects rats' cardiomyocytes from ischaemia/reperfusion-induced apoptosis by affecting the mitochondrial apoptosis pathway
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DOI：
10.1111/apha.12186
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发表时间：
2014-03-01
影响因子：
6.3
通讯作者：
Yao, L-L.
中科院分区：
文献类型：
--
作者：
Wang, Y-G.;Li, Y.;Yao, L-L.
方法采用结扎大鼠左冠状动脉前降支（LAD）30 min，再灌注2 h的方法，建立缺血再灌注模型，观察左旋3丁基苯酞（NBP）对缺血再灌注心肌的保护作用。培养新生心肌细胞并使其缺氧。在体内实验中，在手术前2 h和再灌注后立即腹腔注射L-3-正丁基苯酞;在体外实验中，将L-3-正丁基苯酞加入培养液中。记录血流动力学参数评价心功能，用氯化三苯基四氮唑（TTC）和伊文思蓝（Evens blue）染色确定危险区和梗死区面积，用末端脱氧核苷酸转移酶缺口末端标记（TUNEL）染色计数凋亡细胞数。Western blotting法检测凋亡蛋白水平，免疫组化染色法检测甘油醛-3-磷酸脱氢酶（GAPDH）蛋白转位。结果本研究首次发现，L-3-正丁基苯酞具有明显改善心脏血流动力学功能、减少心肌梗死面积和梗死周边区凋亡细胞数量的作用。细胞凋亡信号研究表明，L-3-正丁基苯酞通过影响线粒体Bcl-2蛋白的表达，抑制caspase 3的激活和细胞色素C的释放，从而影响细胞凋亡信号的传递。结论L-3-正丁基苯酞通过抗氧化作用和影响线粒体凋亡途径，对缺血/再灌注诱导的心肌细胞凋亡具有保护作用。
AimsThis study investigated the role of L-3-n-Butylphthalide (NBP) in cardiac protection.MethodsThe left anterior descending coronary arteries (LAD) of the rats were occluded for 30min following by 2-h reperfusion to make the ischaemia/reperfusion models. Neonatal cardiomyocytes were cultured and subjected to hypoxia. L-3-n-Butylphthalide was administered intraperitoneally 2h before the surgery and right after the reperfusion in the in vivo experiments or added to the culture medium in vitro. Haemodynamic parameters were recorded to evaluate the cardiac functions, triphenyltetrazolium chloride (TTC) and Evens blue staining were used to determine the area of risk and infarct area, apoptotic cell numbers were counted with terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining. Western blotting was used to determine the apoptotic protein levels and immune staining to determine the translocation of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein.ResultsOur research showed for the first time that L-3-n-Butylphthalide had great effects in improving cardiac hemodynamic function and decreasing cardiac infarct areas and apoptotic cell numbers in the peri-infarct areas. The apoptotic signals investigation showed that L-3-n-Butylphthalide affected the mitochondrial pathway including Bcl-2 protein expression, inhibition of caspase 3 activation and cytochrome C releasing. Besides, Glyceraldehyde-3-phosphate dehydrogenase protein translocation was inhibited by L-3-n-Butylphthalide treatment, and this effect was mediated by endogenous reactive oxygen species (ROS).ConclusionL-3-n-Butylphthalide protects cardiomyocytes from ischaemia/reperfusion-induced apoptosis by antioxidant effect and affecting mitochondrial apoptotic pathway.