摘要
目的观察维拉帕米对培养大鼠心肌细胞缺氧复氧(hypoxia/reoxygenation,H/R)所致损伤的作用及其机制。方法制作心肌细胞H/R损伤模型。倒置显微镜和透射电子显微镜下观察心肌细胞形态和结构的变化;双抗体夹心光化学法检测心肌细胞培养上清液中肌钙蛋白(cardiac troponin I,cTnI)的浓度;ELISA方法检测上清液中TNF-α的含量;Western-blot及Northern-blot方法分别检测培养心肌细胞中早期生长反应基因-1(early growth response gene-1,Egr-1)基因和蛋白的表达水平。结果缺氧及复氧过程中给予维拉帕米能改善H/R所致心肌细胞形态结构的损伤,减轻H/R所致cTnI和TNF-α的漏出或分泌,抑制H/R所致的心肌细胞Egr-1基因和蛋白过量表达。结论维拉帕米对培养心肌细胞H/R损伤具有多重保护作用,其机制与其抑制Egr-1基因及蛋白过量表达有关。
Objective To investigate the effect and mechanism of verapamil on rat cardiomyocytes during hypoxia/reoxygenation（H/R） injury.Methods The H/R model of neonatal rat cardiomyocytes was established.The changes of morphology and structure of cardiomyocytes were detected by inverted microscopy and transmission electron microscopy.The levels of cardiac troponin I（cTnI） and tumor necrosis factor-α（TNF-α） in culture medium were measured using chemiluminescence system with two-site sandwich immunoassay kit and ELISA,respectively.The expression levels of early growth response gene-1（Egr-1）mRNA and protein were examined by northern-blot and western-blot,respectively.Results Treatment with verapamil significantly attenuated morphological and structural changes of cardiomyocytes,decreased the cTnI leakage and TNF-α release,and inhibited the overexpression of Egr-1 mRNA and protein in cardiomyocytes during hypoxia/reoxygenation.Conclusion Verapamil can protect cultured cardiomycytes from H/R injury,which is associated with the inhibition of Egr-1 overexpression.
出处
《广东药学院学报》
CAS
2013年第2期172-176,共5页
Academic Journal of Guangdong College of Pharmacy
基金
国家自然科学基金-广东省自然科学基金联合资助基金项目(U0932005)
国家自然科学基金(81072633)
