不同浓度高糖对原代乳鼠心肌细胞及H9C2细胞系线粒体形态影响的观察

The effect of different high glucose concentration on mitochondrial morphology of primary cultured neonatal rat cardiomyocytes and H9C2 cells

目的探讨不同浓度高糖对原代乳鼠心肌细胞及H9C2细胞系线粒体形态及膜电位的影响。方法分离培养原代乳鼠心肌细胞及传代培养H9C2细胞,两种细胞贴壁24 h后分为正常对照组(NC)、甘露醇组(NC+M)、25 mmol/L高糖组(HG25)与35 mmol/L高糖组(HG35),检测各组细胞线粒体形态与线粒体膜电位的变化并测定乳酸脱氢酶(LDH)含量和细胞凋亡率。结果乳鼠心肌细胞及传代培养H9C2细胞中HG25组与HG35组均导致细胞线粒体形态的改变,表现为线粒体面积/周长比及线粒体反圆度降低,两组均使线粒体膜电位水平降低(P<0. 05)。与NC组细胞凋亡率、LDH水平比较,HG25组、NC+M组差异无统计学意义(P>0. 05),HG35组升高,差异有统计学意义(P<0. 05)。结论 25 mmol/L及35 mmol/L两种高糖浓度均能降低心肌细胞线粒体连接度与拉长度,使其由网状向点块状改变,并降低其膜电位水平,影响心肌细胞能量代谢,但25 mmol/L的高糖浓度并未影响心肌细胞凋亡率。

Objective To evaluate the influence of different high glucose concentrations on mitochondrial morphology and membrane potential in primary neonatal rat myocardial cells and H9C2 cell lines.MethodsIsolated primary neonatal rat cardiomyocytes and passaged H9C2 cells were adherent cultured for24 h and then divided into control group(NC),mannitol group(NC + M),25 mmol/L glucose group(HG25)and 35 mmol/L glucose group(HG35). LDH content,cell apoptosis rate,myocardial mitochondrial morphology and mitochondrial membrane potential changes were observed.ResultsAfter treated with25 mmol/L and 35 mmol/L high glucose,neonatal rat myocardial cells and H9C2 cells both exhibited decreased area/perimeter ratio and inverse circularity. Mitochondrial membrane potential decreased significantly in the two high glucose groups(P<0. 05). Cell apoptosis rate and LDH was not statistical different between HG25 group and NC group(P>0. 05). Cell apoptosis rate and LDH were higher in HG35 group(P<0. 05).Cell apoptosis rate and mitochondrial morphology were not influenced in NC+M group.ConclusionHigh glucose with 25 mmol/L and 35 mmol/L concentration reduced myocardial mitochondrial connectivity and length pull,and decreased membrane potential level. Thus affect the myocardial cell energy metabolism due to the mitochondrial morphology changes. But high glucose with 25 mmol/L concentration did not affect myocardial apoptosis rate.