抵挡汤在高糖、高糖波动环境下对人脐静脉血管内皮细胞能量代谢相关因子表达的影响

Effect of Didang Decoction on the expression of energy metabolism related factors in human umbilical vein endothelial cells under high glucose and high glucose fluctuating

关键词:  抵挡汤  高糖波动  能量代谢  p-AMPK  PGC-1α  eNOS  caspase-3

Key words:  Didang Decoction  high glucose fluctuating  energy metabolism  p-AMPK  PGC-1α  eNOS  caspase-3

摘要:

[目的] 通过观察抵挡汤在高糖、高糖波动环境下对人脐静脉血管内皮细胞（HUVECs）磷酸化腺苷酸活化蛋白激酶（p-AMPK）、过氧化物酶体增殖物激活受体γ（PPARγ）共激活因子1α（PGC-1α）、内皮细胞型一氧化氮合酶（eNOS）、半胱氨酸天门冬氨酸特异性蛋白酶3（caspase-3）表达的影响，探讨抵挡汤对血管内皮细胞保护作用。[方法] 模拟高糖、高糖波动环境培养HUVECs并分别进行抵挡汤、辛伐他汀、二甲双胍干预，各组培养48 h后，蛋白免疫印迹（Western Blot）检测p-AMPK、PGC-1α、eNOS蛋白表达，逆转录-聚合酶链式反应（RT-PCR）检测caspase-3基因含量，流式细胞仪检测细胞凋亡率。[结果] 高糖组、高糖波动组p-AMPK、PGC-1α、eNOS表达均显著下降，且以高糖波动组最低。抵挡汤干预后p-AMPK、PGC-1α、eNOS表达均显著上调，与二甲双胍干预无差异。高糖组、高糖波动组caspase-3 mRNA表达均明显升高，抵挡汤干预后均明显降低，且与二甲双胍干预组无差异。[结论] 抵挡汤在高糖、高糖波动环境下均可磷酸化AMPK，上调PGC-1α、eNOS蛋白表达、下调caspase-3基因表达，影响能量代谢和线粒体合成，减少内皮细胞凋亡，从而保护血管内皮。

Abstract:

[Objective] To observe the effect of Didang Decoction (DDD) on the expression of phosphorylated adenylate activated protein kinase (p-AMPK),peroxisome proliferator activated receptor γ co-activated activator 1α (PGC-1α),endothelial nitric oxide synthase (eNOS),caspase-3 in human umbilical vein endothelial cells (HUVECs) under high glucose and high glucose fluctuation environment,the protective effect of DDD on vascular endothelial cells was discussed.[Methods] HUVECs were cultured in a simulated environment with high glucose and high glucose fluctuation and were intervened by DDD,simvastatin,and metformin respectively. After 48 hours of culture in each group,the protein expressions of p-AMPK,PGC-1α and eNOS were detected by Western blot,the gene content of caspase-3 was detected by real-time PCR (RT-PCR),and the apoptosis rate was detected by flow cytometry.[Results] The expression of p-AMPK,PGC-1α and eNOS in high glucose group and high glucose fluctuation group decreased significantly,and the lowest expression was found in high glucose fluctuation group. The expression of p-AMPK,PGC-1α and eNOS protein were significantly up-regulated after the intervention of DDD,and there was no difference with the intervention of metformin. Caspase-3 gene expression in high glucose group and high glucose fluctuation group were significantly increased,and significantly decreased after intervention with DDD,and there was no difference between them and metformin intervention group.[Conclusion] DDD can promote AMPK phosphorylation,up-regulate the expression of PGC-1α and eNOS proteins,reduce caspase-3 gene level,regulate endothelial cell mitochondrial synthesis and energy metabolism,and reduce endothelial cell apoptosis under high glucose and high glucose fluctuation environment,thus protecting vascular endothelium.