高糖微环境对体外心肌细胞增殖活性和氧化应激的影响

Effect of high concentration glucose microenvironment on proliferation activity and oxidative stress in cardiomyocytes in vitro

目的:考察不同浓度葡萄糖对体外心肌细胞增殖活性和氧化应激的影响,探讨糖毒性对心肌细胞的损伤作用及可能的作用机制。方法:将含有不同浓度葡萄糖培养基处理的心肌细胞分别培养24,48,72 h,利用倒置显微镜观察心肌细胞形态学变化,MTT法检测心肌细胞活性,酶标法检测还原型谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)含量,四唑盐(water-soluble tetrazolium,WST-1)法检测超氧化物歧化酶(superoxide dismutase,SOD)活力,流式细胞法检测活性氧(reactive oxygen species,ROS)水平。结果:高浓度葡萄糖(33.30,50.00,100.00mmol·L-1)作用下,心肌细胞形态发生改变。与同期对照组及正常糖浓度组比较,高浓度葡萄糖作用24~72 h后,细胞增殖活性显著降低(P<0.05)。随着葡萄糖浓度增加和作用时间延长,细胞内GSH含量、SOD活力降低,MDA含量和ROS水平升高。结论:高浓度葡萄糖抑制心肌细胞增殖活性,诱导氧化应激反应发生,并通过破坏机体ROS与抗氧化物质之间的平衡,引起心肌细胞损伤。

OBJECTIVE To explore the damage impact and its mechanism of the toxicity of glucose at high concentrations on cardiomocytes,the effects of different concentrations of glucose on proliferation activity and oxidative stress in cardiomyocytes in vitro.METHODS Cardiomyocytes were cultured in the medium containing different concentrations of glucose for 24,48 and72 hours,respectively.The morphological changes of cardiomyocytes were observed by inverted microscope and cell viability was determined by MTT assay.The levels of glutathione（GSH）and malondialdehyde（MDA）were detected by microplate reader.The activity of superoxide dismutase（SOD）was detected by Water-soluble tetrazolium（WST-1）assay.Furthermore,reactive oxygen species（ROS）were detected by flow cytometer.RESULTS The morphological changes of cardiomyocytes were obviously observed in high concentrations of glucose（33.30,50.00,100.00 mmol·L-1）media for 24,48 and 72 hours,respectively.Compared with the low concentration control group and the normal concentration control group at the same treatment time,cell proliferation activity reduced significantly under the high concentration of glucose for 24-72 h（P〈0.05）.With the increase of glucose concentration and the exposure time,the content of GSH and SOD activity were reduced,whereas the MDA and ROS level were increased.CONCLUSION A high concentration of glucose inhibits the proliferation of cardiomyocytes,triggers a series of oxidative stress reactions,and causes damages in H9 c2 cells by destroying the balance between ROS and antioxidants.