摘要
目的研究促红细胞生成素衍生肽(HBSP)对急性缺氧/复氧心肌细胞凋亡的影响及其可能的作用机制。方法体外培养H9C2乳鼠心肌细胞系,建立缺氧(3 h)/复氧(3 h)模型。实验分为对照组、缺氧/复氧组(A/R)、A/R+HBSP组。测定培养细胞上清液乳酸脱氢酶(LDH)含量;用annexin-V与PI双染法及流式细胞仪检测心肌细胞凋亡率;分离细胞质与线粒体,用Western blot法检测线粒体内和胞浆中的细胞色素C蛋白的表达,用caspase试剂盒检测心肌细胞caspase-9及caspase-3的表达。结果与正常对照组比较,A/R组细胞存活率和线粒体内细胞色素C蛋白水平明显下降(P<0.01),而凋亡率、caspase-9、caspase-3活性及胞质内细胞色素C蛋白水平均显著升高(P<0.01)。结论HBSP对缺氧复氧乳鼠心肌细胞具有明显的保护作用,其机制可能与抑制线粒体途径介导的细胞凋亡有关。
Objective To study the effect of Helix B surface peptide（ HBSP） on myocardial apoptosis in the state of acute anoxia / reoxygenation and its possible mechanism. Methods Cultured neonatal rat cardiomyocytes H9C2 cell line in vitro and establishment anoxia（ 3 h） / reoxygenation（ 3 h） model. The experiment was carried by three group： control group,anoxia / reoxygenation group（ A / R） and H / R ＋ HBSP groups. Cultured supernatants were collected to measure the lactate dehydrogenase（ LDH） and the myocardial apoptosis rate was detected by annexinV and PI double staining and flow cytometry. Western blot analysis was used to measure the expression of cytochrome C,and the activity of caspase-3 and caspase-9 was determined by a colorimetric method. Results Compared with the control group,cell viability and mitochondrial cytochrome C protein in A / R group significantly decreased（ P〈0. 01）,while the rate of apoptosis,caspase-9,caspase-3 activity and cytosolic cytochrome C protein levels significantly increased（ P〈0. 01）. Conclusions HBSP significantly protects neonatal rat cardiomyocytes from the injury of anoxia / reoxygenation and the mechanism may be inhibiting the cell apoptotic mediated by mitochondrial pathway.
出处
《基础医学与临床》
CSCD
2016年第1期68-72,共5页
Basic and Clinical Medicine
基金
广东省自然科学基金(2015A030313238)
广东省广州市海珠区科普计划(2014HZKP-HD-35)
广东省医学科研基金(20151261346046)
关键词
HBSP
缺氧/复氧
心肌细胞
凋亡
线粒体途径
HBSP
anoxia/reoxygenation
myocardial cells
apoptosis
mitochondrial pathway
