Thiamine and benfotiamine prevent increased apoptosis in endothelial cells and pericytes cultured in high glucose
Thiamine and benfotiamine prevent increased apoptosis in endothelial cells and pericytes cultured in high glucose
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DOI：
10.1002/dmrr.470
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发表时间：
2004-07-01
影响因子：
8
通讯作者：
Porta, M
中科院分区：
文献类型：
--
作者：
Beltramo, E;Berrone, E;Porta, M
高糖诱导小血管和大血管的病理改变，可能是通过AGE的形成增加、醛糖还原酶和蛋白激酶C的激活以及己糖胺途径的通量增加。我们之前的研究表明，硫胺素和苯甲硫胺可以纠正高葡萄糖内皮细胞的延迟复制并增加乳酸的产生。我们现在的目的是验证在高葡萄糖环境下，硫胺素和苯甲硫胺对内皮细胞和周细胞的细胞周期、凋亡和粘附分子表达的影响。方法人脐静脉内皮细胞和牛视网膜周细胞在正常（5.6 mmol/L）和高（28 mmol/L）葡萄糖环境中培养，添加或不添加硫胺素或苯福地胺，50或100 μ mol/L。凋亡通过两种单独的ELISA方法测定，分别测定DNA片段和caspase-3活性。流式细胞术检测细胞周期和整合素亚单位alpha3、alpha5、beta1的浓度。结果高糖培养3 d后，细胞凋亡增加。在内皮细胞和周细胞。硫胺素和苯甲硫胺逆转了这种作用。在这些实验条件下，细胞周期穿越和整合素浓度都没有改变。结论硫胺素和苯福地胺可纠正高糖诱导的血管细胞凋亡增加。进一步阐明其作用机制有助于为临床使用这种维生素预防和/或治疗糖尿病微血管病变奠定基础。版权所有：John Wiley Sons， Ltd. 2004
Background High glucose induces pathological alterations in small and large vessels, possibly through increased formation of AGE, activation of aldose reductase and protein kinase C, and increased flux through the hexosamine pathway. We showed previously that thiamine and benfotiamine correct delayed replication and increase lactate production in endothelial cells subjected to high glucose. We now aim at verifying the effects of thiamine and benfotiamine on cell cycle, apoptosis, and expression of adhesion molecules in endothelial cells and pericytes, under high ambient glucose.Methods Human umbilical vein endothelial cells and bovine retinal pericytes were cultured in normal (5.6 mmol/L) or high (28 mmol/L) glucose, with or without thiamine or benfotiamine, 50 or 100 mumol/L. Apoptosis was determined by two separate ELISA methods, measuring DNA fragmentation and caspase-3 activity, respectively. Cell cycle and integrin subunits alpha3, alpha5, and beta1 concentration were measured by flow cytometry.Results Apoptosis was increased in high glucose after 3 days of culture, both. in endothelium and pericytes. Thiamine and benfotiamine reversed such effects. Neither cell cycle traversal nor integrin concentrations were modified in these experimental conditions.Conclusions Thiamine and benfotiamine correct increased apoptosis due to high glucose in cultured vascular cells. Further elucidations of the mechanisms through which they work could help set the basis for clinical use of this vitamin in the prevention and/or treatment of diabetic microangiopathy. Copyright (C) 2004 John Wiley Sons, Ltd.