津力达联合通心络减轻高糖环境下肾脏微血管内皮细胞内质网凋亡

Effect of Jinlida combined with Tongxinluo on apoptosis of renal microvascular endothelial cells in high-glucose environment

目的:观察津力达联合通心络对高糖环境下肾脏微血管内皮细胞内质网凋亡途径的影响,探讨其减轻肾脏微血管内皮细胞损伤的作用机制。方法:将体外培养的肾脏微血管内皮细胞分为正常对照组、高糖组、通心络组、津力达组和通心络+津力达组。干预24 h后,CCK-8法检测细胞活力;流式细胞术检测细胞凋亡率和活性氧簇(ROS)水平。Western blot检测蛋白激酶R样内质网激酶(PERK)、磷酸化PERK(p-PERK)、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、caspase-12和Bcl-2的蛋白水平。结果:与正常对照组比较,高糖环境下肾脏微血管内皮细胞活力降低,细胞凋亡率增加,ROS生成增多,PERK、p-PERK、GRP78、CHOP和caspase-12的蛋白水平升高,而Bcl-2的蛋白水平下降(P<0.05)。与高糖组比较,通心络、津力达和通心络+津力达均能不同程度地增强高糖环境下肾脏微血管内皮细胞活力,减少细胞凋亡和ROS生成,抑制PERK、p-PERK、GRP78、CHOP和caspase-12蛋白的水平,提高Bcl-2蛋白的水平(P<0.05)。与津力达组和通心络组比较,通心络+津力达组各指标的差异均有统计学意义(P<0.05)。结论:津力达和通心络均能够减轻高糖环境下肾脏微血管内皮细胞损伤,其机制与减轻内质网应激介导的凋亡途径有关;两者联合应用能够协同增强肾脏微血管内皮细胞保护作用。

AIM:To observe the effect of Jinlida combined with Tongxinluo on the apoptosis of renal microvascular endothelial cells in the high-glucose environment,and to explore their mechanism of protecting renal microvascular endothelial cells.METHODS:The renal microvascular endothelial cells cultured in vitro were divided into control group,high glucose group,Tongxinluo group,Jinlida group,and Tongxinluo+Jinlida group. After intervention for 24 h,CCK-8 assay was used to measurethe cell viability. The apoptotic rate and reactive oxygen species(ROS)level were measured by flow cytometry. Western blot was used to determine the protein levels of protein kinase R-like endoplasmic reticulum kinase(PERK),phosphorylated PERK(p-PERK),glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),caspase-12 and Bcl-2.RESULTS:Compared with control group,the viability of renal microvascular endothelial cells in the high-glucose environment was decreased,the apoptotic rate,the ROS level and the protein levels of PERK,p-PERK,GRP78,CHOP and caspase-12 were increased,while Bcl-2 protein was decreased(P<0. 05). In comparison with high glucose group,the viability of renal microvascular endothelial cells in Tongxinluo,Jinlida and Tongxinluo+Jinlida groups was increased to varying degrees,the apoptotic rate,the ROS level and the protein levels of PERK,pPERK,GRP78,CHOP and caspase-12 were decreased,while Bcl-2 protein was increased(P<0. 05). Compared with Jinlida group and Tongxinluo group,the improvement of each index in Jinlida+Tongxinluo group was statistically significant.CONCLUSION:Jinlida and Tongxinluo attenuate the damage of renal microvascular endothelial cells in the highglucose environment,and the mechanism may be related to the reduction of endoplasmic reticulum stress-mediated apoptosis pathway. The combined application of Jinlida and Tongxinluo synergistically enhances the protective effect of the drugs on the renal microvascular endothelial cells.