High D-glucose-induced changes in endothelial Ca2+/EDRF signaling are due to generation of superoxide anions
High D-glucose-induced changes in endothelial Ca2+/EDRF signaling are due to generation of superoxide anions
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DOI：
10.2337/diabetes.45.10.1386
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发表时间：
1996-10-01
期刊：
影响因子：
7.7
通讯作者：
Kostner, GM
中科院分区：
文献类型：
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作者：
Graier, WF;Simecek, S;Kostner, GM
用高D-葡萄糖预处理猪主动脉内皮细胞，由于内皮Ca 2+释放（57%）和Ca 2+进入缓激肽（97%）增加，导致内皮源性舒张因子（EDRF）形成增加（39%）。本研究旨在探讨高糖影响内皮细胞Ca ~（2+）/EDRF反应的细胞内机制。醛糖还原酶抑制剂，sorbinil和zopolrestat，未能减少高D-葡萄糖介导的Ca 2 +/EDRF反应的变化，表明醛糖还原酶不有助于高D-葡萄糖引发的Ca 2 +/EDRF信号的变化。用非代谢性D-葡萄糖类似物3-O-甲基吡喃葡萄糖（3-OMG）预处理细胞，模拟了高D-葡萄糖对钙释放（41%）和钙进入缓激肽（114%）的影响，与EDRF形成升高（26%）相关。高糖和3-OMG增加超氧阴离子（O-2（-））的生成（分别为133%和293%），对环氧合酶抑制剂不敏感（5，8，11，14-二十碳四炔酸[ETYA]，吲哚美辛），脂氧合酶（ETYA、棉酚、去甲二氢愈创木酸[NDGA]）、细胞色素P450（NDGA、益康唑、咪康唑）和一氧化氮（NO）合酶（L-N-Omega-nitroarginine），而去铁醛（desferal），一种金属螯合剂，减少了这种作用。γ-谷氨酰-半胱氨酸合成酶抑制剂丁硫氨酸磺酰亚胺（BSO）也使O-2（-）的形成增加了365%，并模拟了高D-葡萄糖对Ca 2 +/EDRF信号传导的影响。高D-葡萄糖，3-OMG，和BSO的影响被取消与超氧化物歧化酶共孵育。与高D-葡萄糖一样，用O-2（-）-生成系统预处理，黄嘌呤氧化酶/次黄嘌呤，缓激肽刺激的Ca 2+释放（+10%），Ca 2+内流（+75%）和EDRF（+73%）升高。我们认为，长期暴露于病理性高浓度的D-葡萄糖导致O-2（-）的形成增强，可能是由于金属介导的D-葡萄糖在细胞内的氧化。这种O-2（-）的过冲通过一种未知的机制增强激动剂刺激的Ca 2 +/EDRF信号传导。
Pretreatment of porcine aortic endothelial cells with high D-glucose results in enhanced endothelium-derived relaxing factor (EDRF) formation (39%) due to increased endothelial Ca2+ release (57%) and Ca2+ entry (97%) to bradykinin. This study was designed to investigate the intracellular mechanisms by which high D-glucose affects endothelial Ca2+/EDRF response. The aldose-reductase inhibitors, sorbinil and zopolrestat, failed to diminish high D-glucose-mediated alterations in Ca2+/EDRF response, suggesting that aldose-reductase does not contribute to high D-glucose-initiated changes in Ca2+/EDRF signaling. Pretreatment of cells with the nonmetabolizing D-glucose analog, 3-O-methylglucopyranose (3-OMG), mimicked the effect of high D-glucose on Ca2+ release (41%) and Ca2+ entry (114%) to bradykinin, associated with elevated EDRF formation (26%). High D-glucose and 3-OMG increased superoxide anion (O-2(-)) formation (133 and 293%, respectively), which mas insensitive to inhibitors of cyclooxygenase (5,8,11,14-eicosatetraynoic acid [ETYA], indomethacin), lipoxygenase (ETYA, gossypol, nordihydroguaiaretic acid [NDGA]), cytochrome P450 (NDGA, econazole, miconazole), and nitric oxide (NO) synthase (L-N-Omega-nitroarginine), while it was diminished by desferal, a metal chelator. The gamma-glutamyl-cysteine-synthase inhibitor, buthioninesulfoximine (BSO), also increased formation of O-2(-) by 365% and mimicked the effect of high D-glucose on Ca2+/EDRF signaling. The effects of high D-glucose, 3-OMG, and BSO were abolished by co-incubation with superoxide dismutase. Like high D-glucose, pretreatment with the O-2(-)-generating system, xanthine oxidase/hypoxanthine, elevated bradykinin-stimulated Ca2+ release (+10%), Ca2+ entry (+75%), and EDRF (+73%). We suggest that prolonged exposure to pathologically high D-glucose concentration results in enhanced formation of O-2(-), possibly due to metal-mediated oxidation of D-glucose within the cells. This overshoot of O-2(-) enhances agonist-stimulated Ca2+/EDRF signaling via a yet unknown mechanism.