Acute hyperglycemia induces nitrotyrosine formation and apoptosis in perfused heart from rat
Acute hyperglycemia induces nitrotyrosine formation and apoptosis in perfused heart from rat
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DOI：
10.2337/diabetes.51.4.1076
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发表时间：
2002-04-01
期刊：
影响因子：
7.7
通讯作者：
Giugliano, D
中科院分区：
文献类型：
--
作者：
Ceriello, A;Quagliaro, L;Giugliano, D
本研究探讨了冠状动脉灌注压，一氧化氮（NO）和超氧化物的生产，硝基酪氨酸（NT）的形成，心肌细胞凋亡在离体心脏灌注高浓度葡萄糖。冠状动脉灌注压; NO和超氧阴离子的产生; NT、诱导型NO合酶（iNOS）和组成型NO合酶（NOS）eNOS的免疫组化染色;在用含有浓度为11.1mmol/l的D-葡萄糖的溶液（对照）灌注2小时的心脏中研究心脏细胞的凋亡，浓度为33.3 mmol/l的D-葡萄糖（高糖），或D-葡萄糖（33.3 mmol/l）加谷胱甘肽（0.3 mmol/l）。在高糖条件下灌流离体心脏，可引起冠脉灌注压显著升高（P < 0.001），NO和超氧化物生成增加。然而，超氧化物的产生比基线高300%，而NO的产生高40%（两者P < 0.001）。这种作用伴随着NT的形成和iNOS表达的增加。eNOS保持不变。在实验结束时，心脏细胞凋亡是明显的心脏灌注与高葡萄糖。谷胱甘肽可显著预防高糖的影响。本研究表明高糖2 h足以增加工作大鼠心脏iNOS基因表达和NO释放。iNOS的上调和NO生成的增加伴随着超氧化物产生的显著伴随增加，这是一种有利于过氧亚硝酸盐产生的条件，过氧亚硝酸盐是一种强大的促氧化剂，可以通过自身和/或通过形成硝基酪氨酸介导高葡萄糖对心脏的毒性作用，如细胞凋亡检测所示。
This study investigated coronary perfusion pressure, nitric oxide (NO) and superoxide production, nitrotyrosine (NT) formation, and cardiac cell apoptosis in isolated hearts perfused with high glucose concentration. Coronary perfusion pressure; NO and superoxide anion generation; immunostaining for NT, inducible NO synthase (iNOS), and the constitutive type of NO synthase (NOS) eNOS; iNOS and eNOS mRNA expression by Western blot and RT-PCR; and apoptosis of cardiac cells were studied in hearts perfused for 2 h with solutions containing D-glucose at a concentration of 11.1 mmol/l (control), D-glucose at the concentration of 33.3 mmol/l (high glucose), or D-glucose (33.3 mmol/l) plus glutathione (0.3 mmol/l). Perfusion of isolated hearts in conditions of high glucose concentration caused a significant increase of coronary perfusion pressure (P < 0.001) and an increase of both NO and superoxide generation. However, superoxide production was 300% higher than baseline, whereas NO production was 40% higher (P < 0.001 for both). This effect was accompanied by the formation of NT, and an increase of iNOS expression. eNOS remained unchanged. At the end of the experiments, cardiac cell apoptosis was evident in hearts perfused with high glucose. The effects of high glucose were significantly prevented by glutathione. This study demonstrates that high glucose for 2 h is enough to increase iNOS gene expression and NO release in working rat hearts. Upregulation of iNOS and raised NO generation are accompanied by a marked concomitant increase of superoxide production, a condition favoring the production of peroxynitrite, a powerful pro-oxidant that can mediate the toxic effects of high glucose on heart by itself and/or via the formation of nitrotyrosine, as suggested by the detection of cell apoptosis.