Zhao Yuling, Lu Naihao, Li Hailing, Zhang Yan, Gao Zhonghong, Gong Yuefa
Department of Chemistry and Chemical Engineering, Huazhong University of Science & Technology, Wuhan 430074, PR China.
Mol Cell Biochem. 2008 Apr;311(1-2):19-29. doi: 10.1007/s11010-007-9688-y. Epub 2007 Dec 14.
The dysfunction and further damage of endothelium play an important role in the development and progression of diabetic vascular complications. Protein tyrosine nitration is involved in endothelial cell injury induced by high glucose. Little is known about protein nitration in human umbilical vein endothelial cells (ECV304) induced by high glucose. In the present article, exposure of ECV304 to 30 mM high glucose (HG30) and 40 mM high glucose (HG40) or hemin-nitrite-H2O2 system for 72 h, the cell injury in ECV304 induced by high glucose and exogenous nitrating agent was studied. After 72 h treatment, it was found that high glucose stimulated ECV304 injury in a dose-dependent manner, including reducing cell viability, increasing malondialdehyde (MDA) content, decreasing glutathione (GSH) content, increasing intracellular reactive oxygen species (ROS), increasing the production of nitric oxygen (NO) (increased nitrite content in cell and nitrate content in medium) and generating protein tyrosine nitration. It was also found that protein tyrosine nitration could induce cell injury further. By comparison the protein tyrosine nitration induced by high glucose condition and extrinsic factors (hemin-nitrite-H2O2 system), it may be speculated that protein is nitrated selectively to generate nitrotyrosine in diabetic vascular complications.
内皮功能障碍及进一步损伤在糖尿病血管并发症的发生和发展中起重要作用。蛋白质酪氨酸硝化参与高糖诱导的内皮细胞损伤。关于高糖诱导人脐静脉内皮细胞（ECV304）中的蛋白质硝化知之甚少。在本文中，将ECV304暴露于30 mM高糖（HG30）、40 mM高糖（HG40）或血红素-亚硝酸盐-H2O2体系72小时，研究了高糖和外源性硝化剂诱导的ECV304细胞损伤。处理72小时后，发现高糖以剂量依赖方式刺激ECV304损伤，包括降低细胞活力、增加丙二醛（MDA）含量、降低谷胱甘肽（GSH）含量、增加细胞内活性氧（ROS）、增加一氧化氮（NO）生成（细胞内亚硝酸盐含量和培养基中硝酸盐含量增加）以及产生蛋白质酪氨酸硝化。还发现蛋白质酪氨酸硝化可进一步诱导细胞损伤。通过比较高糖条件和外源性因素（血红素-亚硝酸盐-H2O2体系）诱导的蛋白质酪氨酸硝化，推测在糖尿病血管并发症中蛋白质被选择性硝化以生成硝基酪氨酸。
