高糖高脂对培养成年大鼠心肌细胞损伤观察及机制初探

目的建立成年大鼠心肌细胞培养模型，观察给予高糖、高脂刺激后是否发生心肌细胞损伤和凋亡，并分析其可能机制。方法将分离所得成年大鼠心肌细胞接种入培养皿后随机分为2组进行培养。正常对照组：以M199培养基（Media199）加入5mmol／L葡萄糖与20mmol／L甘露醇作为正常对照培养基培养细胞；高糖高脂组：以M199加入高糖（25mmol／L葡萄糖）高脂（600μmol／L软脂酸）作为培养基培养模拟糖尿病。培养48h后，分别收集培养基上清与细胞，进行心肌损伤和凋亡指标测定。结果高糖高脂组乳酸脱氢酶（LDH），细胞凋亡蛋白caspase-3、caspase-8、caspase-9活性显著升高，与正常对照组比较，差异均具有统计学意义（P均〈0．01）。结论在培养的成年大鼠心肌细胞，高糖、高脂可引起心肌细胞损伤和凋亡，这种凋亡与easpase-8和caspase-9依赖的凋亡途径有关。

Research of mechanisms of injury in adult rat cardiomyocytes by high glucose and high fatty acid culture

Objective To observe the injury and apoptosis in adult rat cardiomyocytes induced by high glucose and high fatty acid culture, and to investigate its possible mechanisms. Methods Cardiomyocytes were isolated from adult rats. The cell culture was randomly divided into two groups. Control group： cultured by M199 with 5 mmol/L glucose and 20 mmol/L mannitol. High glucose and high fatty acid group,cultured by M199 with 25 mmol/L glucose and 600 μmol/L palmitate. Cardiomyocytes were cultured in those two kinds of media for 48 hours and then the conditioned media and cardiomyocytes were collected respectively for different assays. Results Compared with Control group, high glucose and high fatty acid group showed increased activity of LDH which indicated the injury of cardiomyocytes, and also increased activity of caspase-3, caspase-8 and caspase-9 which implied for apoptosis of cardiomyocytes. Conclusion High glucose and high fatty acid could cause the injury and apoptosis in cultured adult rat cardiomyocytes. This cardiomyocyte apoptosis was mainly induced through caspase- 8-and caspase-9-dependent pathways.