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目的研究丙酮酸脱氢酶激动剂二氯醋酸二异丙胺(DADA)对人脐静脉内皮细胞(HUVECs)的功能的影响及可能机制。方法以体外培养的人脐静脉内皮细胞(HUVECs)作为靶细胞,进行了以下研究:1.DADA对高糖诱导的血管EC功能指标一氧化氮(NO)和可溶性细胞间黏附分子-1(sICAM-1)的影响;2.用激光共聚焦显微镜及Westernblotting杂交分别观察高糖诱导的PKCα、PKCδ的位置变化及蛋白表达量的影响;实验分组:对照组:浓度为5.5mM;高糖组:浓度为30mM;DADA(1×10-5M,1×10-4Mor1×10-3M)+高糖组。结果1.30mM高糖可诱导EC功能紊乱:高糖可诱导EC功能紊乱:NO浓度降低,sICAM-1水平增加,DADA可抑制高糖诱导的上述变化;高糖可诱导HUVECs的PKCα及PKCδ表达位置转移;PKCδ表达增强,但对PKCα表达影响不明显。结论DADA可以纠正高糖所致内皮细胞功能紊乱,可能部分是通过PKCα及PKCδ途径来实现的。
Objective To investigate the effect of pyruvate dehydrogenase agonist DADA on the function of human umbilical vein endothelial cells (HUVECs) and its possible mechanism. Methods Human umbilical vein endothelial cells (HUVECs) cultured in vitro were used as target cells. The following studies were carried out: 1. DADA was conducted on high glucose-induced vascular endothelial growth factor (NO) and soluble intercellular adhesion molecule-1 -1) .2. The effects of high glucose on PKCα, PKCδ and protein expression were observed by confocal laser scanning microscopy and Western blotting. The experimental group: control group: 5.5mM; Concentration of 30mM; DADA (1 × 10-5M, 1 × 10-4Mor1 × 10-3M) + high glucose group. Results 1.30mM high glucose can induce EC dysfunction: high glucose can induce EC dysfunction: NO concentration decreased, sICAM-1 levels increased, DADA can inhibit high glucose-induced changes; high glucose can induce HUVECs expression of PKCα and PKCδ Metastasis; PKCδ expression increased, but the impact of PKCα expression was not obvious. Conclusion DADA can correct endothelial dysfunction caused by high glucose, probably through PKCα and PKCδ pathways.