[Objective] The aim of this study was to determine whether diisopropylamine dichloroacetate (DADA), a pyruvate dehydrogenase activator, has protective effects on functions of human umbilical vein endothelial cells (HUVECs) and the underlying mechanisms. [Methods] HUVECs were cultured in medium containing 5.5 mM or 30 mM glucose with or without various concentrations (1×10-5 M, 1×10-4 M or 1×10-3 M) of DADA. Expression of nitric oxide (NO) and soluble intercellular adhesion molecule-1 (sICAM-1) was determined by commercially available kits. Distribution and expression of PKCα and PKCδ were determined by laser-scanning confocal microscopy and by Western blot, respectively. [Results] Glucose at 30 mM decreased NO expression but increased sICAM-1 expression. However, these cellular dysfunctions were reversed to the normal levels by co-treatment with DADA.Moreover, DADA inhibited the translocation of PKCα from the plasma to the nucleus and the translocation of PKCδ from the plasma towards the membrane induced by high glucose in HUVECs, and inhibited the high glucose-induced overexpression of PKCδ. [Conclusions] Our findings indicate that DADA corrects high glucose-induced endothelial cell dysfunctions, probably by regulating the PKCα and PKCδ signaling pathways.%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Western blotting杂交分别观察高糖诱导的PKCα、PKC δ的位置变化及蛋白表达量的影响;实验分组:对照组:浓度为5.5 mM;高糖组:浓度为30 mM;DADA(1×10-5 M,1×10-4 M or 1×10-3 M)+高糖组.结果 1.30 mM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PKC δ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.
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