Pro-oxidant role of heme oxygenase in mediating glucose-induced endothelial cell damage
Pro-oxidant role of heme oxygenase in mediating glucose-induced endothelial cell damage
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DOI：
10.1080/10715760400017228
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发表时间：
2004-12-01
影响因子：
3.3
通讯作者：
Chakrabarti, S
中科院分区：
文献类型：
--
作者：
Chen, SL;Khan, ZA;Chakrabarti, S
血管内皮细胞的氧化损伤可能在慢性糖尿病并发症中介导葡萄糖诱导的细胞功能障碍中起关键作用。本研究旨在阐明葡萄糖诱导的高诱导血红素加氧酶（HO）的改变在介导血管内皮细胞氧化应激中的作用。我们还研究了HO和一氧化氮（NO）系统之间的相互作用，以及它在其他血管活性因子改变中的可能作用。人脐静脉内皮细胞（HUVECs）暴露于低（5 mmol/l）和高（25 mmol/l）葡萄糖水平。为了确定HO在内皮功能障碍中的作用并阐明HO和NO系统之间可能的相互作用，将细胞暴露于HO诱导剂（氯化血红素，10 μ mol/l）、HO拮抗剂（SnPPIX，10 μ mol/l）和NO合酶阻断剂（L-NAME，200 μ mol/l），有或没有NO供体（精氨酸，1 mmol/l）。采用真实的时间RT-PCR检测HO和NO亚型的mRNA表达。通过胆红素产生测量HO活性，通过8-羟基-2 '-脱氧鸟苷（8-OHdG）和硝基酪氨酸染色评估细胞氧化应激。葡萄糖诱导内皮细胞HO-1表达上调，HO活性增强，HO与NO之间存在协同刺激关系。HO活性增加也与内皮细胞中的氧化DNA和蛋白质损伤有关。此外，HO活性增加的血管活性因子，ET-1和VEGF的mRNA表达。这些数据表明，HO本身和通过其他血管活性因子的加工可能会导致内皮损伤和功能改变。这些发现在慢性糖尿病并发症的背景下是重要的。
Oxidative damage to the vascular endothelial cells may play a crucial role in mediating glucose-induced cellular dysfunction in chronic diabetic complications. The present study was aimed at elucidating the role of glucose-induced alteration of highly inducible heme oxygenase (HO) in mediating oxidative stress in the vascular endothelial cells. We have also investigated the interaction between HO and the nitric oxide (NO) system, and its possible role in alteration of other vasoactive factors.Human umbilical vein endothelial cells (HUVECs) were exposed to low (5 mmol/l) and high (25 mmol/l) glucose levels. In order to determine the role of HO in endothelial dysfunction and to elucidate a possible interaction between the HO and NO systems, cells were exposed to HO inducer (hemin, 10 mumol/l), HO antagonist (SnPPIX, 10 mumol/l), and NO synthase blocker (L-NAME, 200 mumol/l) with or without NO donor (arginine, 1 mmol/l). mRNA expression of HO and NO isoforms was measured by real time RT-PCR. HO activity was measured by bilirubin production and cellular oxidative stress was assessed by 8-hydroxy-2'-deoxyguanosine (8-OHdG) and nitrotyrosine staining. We also determined the expression of vasoactive factors, endothelin-1 (ET-1) and vascular endothelial growth factor (VEGF).In the endothelial cells, glucose caused upregulation of HO-1 expression and increased HO activity A co-stimulatory relationship between HO and NO was observed. Increased HO activity also associated with oxidative DNA and protein damage in the endothelial cells. Furthermore, increased HO activity augmented mRNA expression of vasoactive factors, ET-1 and VEGF. These data suggest that HO by itself and via elaboration of other vasoactive factors may cause endothelial injury and functional alteration. These findings are of importance in the context of chronic diabetic complications.