摘要
目的观察甘氨酰谷氨酰胺(Gly-Gln)对缺氧/复氧(H/R)损伤后心肌细胞凋亡、线粒体膜电位及心肌酶释放的影响,探讨Gly-Gln对心肌H/R损伤的防治作用及机制。方法利用原代培养的SD大鼠乳鼠心肌细胞建立体外心肌H/R模型,实验分为Control组,H/R组,H/R+Gly-Gln(1,4,16 mmol·L-1)组,Gly-Gln(1,4,16 mmol·L-1)组。采用MTT法测定各组心肌细胞存活率;并检测各组培养液中乳酸脱氢酶(LDH)、肌酸激酶(CK)含量;以Annexin V联合PI染色法检测各组心肌细胞凋亡率;以JC-1为荧光分子探针检测各组心肌细胞线粒体膜电位(△Ψm)变化。结果 H/R组心肌细胞活力低于对照组(P<0.01),而培养液中心肌细胞释出的LDH、CK含量则高于对照组(P<0.01);H/R+Gly-Gln组心肌细胞活力高于H/R组(P<0.01),而培养液中LDH、CK含量则低于H/R组(P<0.05),其效应在一定浓度范围内呈剂量依赖关系。H/R组心肌细胞凋亡率升高,线粒体膜电位明显低于对照组(P<0.01);H/R+Gly-Gln组心肌细胞凋亡率较H/R组降低(P<0.05),且心肌细胞线粒体膜电位升高,与H/R组比较差异有显著性(P<0.01)。结论 Gly-Gln可对抗H/R损伤,发挥细胞保护作用,维持心肌细胞活力,减少心肌细胞H/R损伤所致心肌酶(LDH、CK)释放,并可抑制心肌细胞凋亡,具体机制可能与其稳定心肌细胞线粒体膜电位等有关。
Aim To study the effect and its mechanism of Glycyl-L-glutamine（Gly-Gln） on mitochondrial membrane potentials,apoptosis,and the release of myocardial enzymes of cardiomyocytes suffering from hypoxia/reoxygenation（H/R） injury.Methods A cardiac H/R model in vitro was established with primary cultured neonate rat cardiomyocytes.There were some experimental groups：（1）control group,（2）H/R gr-oup,（3）H/R＋Gly-Gln（1,4,16 mmol·L^-1） group,（4）Gly-Gln（1,4,16 mmol·L^-1） group.The viability of cardiomyocytes was observed by MTT assay.The contents of lactate dehydrogenase（LDH） and creatine kinase（CK） in the culture solution of various groups were determined.The apoptotic rate of cardiomyocytes was analysed by AnnexinV/PI double staining.The mitochondrial membrane potential（△Ψm） of cardiomyocytes was measured by JC-1 as a fluorescent molecular probe.Results Gly-Gln increased remarkably viability of cardiomyocytes suffering from H/R injury（P〈0.01）,inhibited the release of LDH and CK which induced by H/R injury（P〈0.05）.This effect of Gly-Gln was in a dose-dependent manner within certain concentration range.Gly-Gln reduced the apoptotic rate of cardiomyocytes（P〈0.01）.The △Ψm of cardiomyocyte was lower in H/R group than that in control group（P〈0.01）.But the △Ψm was higher in H/R＋ Gly-Gln group than that in H/R group（P〈0.01）.Conclusion Gly-Gln promotes the survival of cardiomyocytes subjecting to H/R injury,reduces the release of myocardial enzymes induced by H/R injury,and prevents the apoptosis and decrement in mitochondrial membrane potential of cardiomyocytes suffering from H/R injury.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第5期656-661,共6页
Chinese Pharmacological Bulletin
基金
广东省自然科学基金博士科研启动项目(No9451063201002960)
中央高校基本科研业务费专项资金资助(No21610707)
关键词
甘氨酰谷氨酰胺
缺血/再灌注
心肌
细胞培养
凋亡
线粒体膜电位
Glycyl-L-glutamine
ischemia/reperfusion
myocardium
cell culture
apoptosis
mitochondrial membrane potential
