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High glucose induced human umbilical vein endothelial cell injury: involvement of protein tyrosine nitration

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Abstract

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.

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Abbreviations

GSH:

Glutathione

HG30:

30 mM high glucose

HG40:

40 mM high glucose

MDA:

Malondialdehyde

NG:

Normal glucose

NO:

Nitric oxygen

NT:

Nitrotyrosine

PGI2 :

Prostaglandin I2

PI3:

Phosphatidyl inositol 3

ROS:

Reactive oxygen species

S.D:

Standard deviation

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 30300073), the Program for New Century Excellent Talents in University (No. NCET-05-0649) and E-Institutes of Shanghai Municipal Education Commission (No.E-04010).

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Authors and Affiliations

  1. Department of Chemistry and Chemical Engineering, Huazhong University of Science & Technology, Wuhan, 430074, P.R. China

    Yuling Zhao, Naihao Lu, Hailing Li, Yan Zhang, Zhonghong Gao & Yuefa Gong

  2. Hubei Key Laboratory of Bioinorganic Chemistry & Materia Medica, Huazhong University of Science & Technology, Wuhan, 430074, P.R. China

    Yuling Zhao, Naihao Lu, Hailing Li, Yan Zhang, Zhonghong Gao & Yuefa Gong

  3. E-Institutes of Shanghai Universities, Division of Nitric Oxide and Inflammatory Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, P.R. China

    Zhonghong Gao

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  1. Yuling Zhao
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Correspondence to Zhonghong Gao.

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Zhao, Y., Lu, N., Li, H. et al. High glucose induced human umbilical vein endothelial cell injury: involvement of protein tyrosine nitration. Mol Cell Biochem 311, 19–29 (2008). https://doi.org/10.1007/s11010-007-9688-y

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  • DOI: https://doi.org/10.1007/s11010-007-9688-y

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