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Mechanistic Studies of AVE3085 Against Homocysteine in Endothelial Protection

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Abstract

Purpose

Homocysteine (Hcy) is an independent risk factor for cardiovascular diseases that impairs endothelial function. We investigated whether the impaired endothelial function can be restored by the eNOS transcription enhancer AVE3085 in porcine coronary arteries. The effects of AVE3085 against Hcy on eNOS-NO function were studied and further investigations were conducted to reveal the role of arginase and the signaling pathway of eNOS activation in the effect of AVE3085 on endothelial dysfunction caused by Hcy.

Methods

Myograph study of vasorelaxation, electrochemical measurement of NO, RT-PCR and Western blot analysis of eNOS, iNOS expression, and eNOS phosphorylation were performed. Arginase activity was determined by urea production and O2 .− generation by lucigenin-enhanced chemiluminenscence.

Results

Exposure to Hcy for 24 h attenuated bradykinin-induced relaxation and NO release, downregulated eNOS mRNA expression and protein expressions of eNOS and p-eNOSSer1177 whereas it upregulated iNOS expression. AVE3085 restored NO release and relaxation, enhanced eNOS but decreased iNOS expression. Inhibition of protein kinase Akt or PI3 kinase attenuated the effect of AVE3085 on relaxation and eNOS phosphorylation. Arginase activity and O2 .− production were inhibited by AVE3085 in Hcy-exposed vessels.

Conclusions

AVE3085 prevents Hcy-induced endothelial dysfunction in coronary arteries by preservation of NO production and suppression of O2 .− generation. Preservation of NO is attributed to upregulation of eNOS expression, activation of eNOS via phosphorylation of Ser1177 through a PI3 kinase/Akt-dependent pathway, and inhibition of arginase. Reduction of O2 .− generation results from reversal of eNOS uncoupling and inhibition of arginase and iNOS.

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Acknowledgments

This work was supported by Hong Kong GRF grants CUHK4789/09M & 4774/12M, National Natural Science Foundation of China 81200123, CUHK Direct Grant 4054015, Tianjin Municipal Science & Technology Commission (10JCYBJC26400), and Key Medical Program of Tianjin Binhai New Area Health Bureau (2011BHKZ001 & 2012BWKZ008) and Tianjin Health Bureau (2012KZ009).

The authors gratefully acknowledge the generous provision of AVE3085 by Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany.

Conflict of Interest

None.

Author information

Authors and Affiliations

  1. Division of Cardiology, Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Hong Kong & TEDA International Cardiovascular Hospital, Medical College, Nankai University, Tianjin, China

    Qin Yang

  2. Department of Physiology, Hebei Medical University, Hebei, China

    Hong-Mei Xue

  3. Division of Cardiothoracic Surgery, Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, Hong Kong

    Malcolm John Underwood

  4. Division of Cardiology, Department of Medicine and Therapeutics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, Hong Kong

    Cheuk-Man Yu

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  1. Qin Yang
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  2. Hong-Mei Xue
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  4. Cheuk-Man Yu
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Corresponding author

Correspondence to Qin Yang.

Additional information

Qin Yang and Hong-Mei Xue contribute equally.

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Yang, Q., Xue, HM., Underwood, M.J. et al. Mechanistic Studies of AVE3085 Against Homocysteine in Endothelial Protection. Cardiovasc Drugs Ther 27, 511–520 (2013). https://doi.org/10.1007/s10557-013-6478-5

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