摘要
目的:探讨氧化应激在高糖诱导乳鼠心肌细胞凋亡中的作用。方法:分离培养SD乳鼠心肌细胞,分别用10～50mmol/L终浓度的葡萄糖进行干预,MTT测定各组心肌细胞的生长活力;激光共聚焦显微镜、脱氧核糖核酸转移酶原位缺口末端标记法(TUNEL)和流式细胞术用于观察和检测心肌细胞凋亡;将心肌细胞分为3组:对照组、高糖组和抗氧化剂组,分别检测各组上清液中乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)的含量以反映心肌细胞的氧化应激水平。结果:MTT结果显示高糖能浓度依赖性地抑制心肌细胞的生长活力;激光共聚焦显微镜、TUNEL染色和流式细胞术均表明高糖能诱导心肌细胞凋亡;与对照组相比,高糖组LDH、MDA含量明显增高,SOD含量显著下降,与高糖组相比,抗氧化剂组LDH,MDA含量明显下降,SOD含量显著升高;流式细胞检测显示高糖组细胞凋亡率较对照组明显增加,而抗氧化剂组细胞凋亡率较高糖组则显著下降。结论:氧化应激是高糖诱导心肌细胞凋亡的一个重要机制。
Objective：To investigate the effect of oxidative stress on high glucose-induced apoptosis in neonatal rat cardiomyocytes. Methods：MTT assay was carried out to detect the viability of cultrured cardiomyocytes exposed to different concentrations of glucose ranging from 10 mmol / L to 50 mmol / L. Laser confocal microscopy,TUNEL assay and flow cytometry were designed to observe and detect cellular apoptosis. Cardiomyocytes were randomly divided into three groups：the control group,the high glucose group and the antioxidant group. Oxidative stress was evaluated by detecting levels of LDH,MDA and SOD in the supernatant of culture media. Results：High glucose was found to inhibit cardiomyocyte viability in a concentration-dependent manner. The results of laser confocal microscopy,TUNEL assay and flow cytometry all suggested that high glucose can induce cardiomyocyte apoptosis. Our findings showed significant increase of LDH and MDA levels and decrease of SOD activity in the high glucose group compared with the control group. However,in the antioxidant group,LDH and MDA levels decreased and SOD activity increased significantly compared with those in the high-glucose group. The apoptotic rate of cardiomyocytes in the high glucose group increased significantly compared with that in the control group,while the apoptotic rate in the antioxidant group declined remarkably in comparison with that in the high glucose group. Conclusion：Oxidative stress is an important mechanism for high glucose-induced cardiomyocyte apoptosis.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第3期345-349,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省"科教兴卫工程"医学重点学科基金
江苏省高校自然科学基金项目
关键词
氧化应激
高糖
心肌细胞
凋亡
oxidative stress
high glucose
cardiomyocyte
apoptosis
