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Cardiovascular Research 1999 41(1):109-115; doi:10.1016/S0008-6363(98)00196-5
© 1999 by European Society of Cardiology
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Copyright © 1999, European Society of Cardiology

Modulation of constitutive nitric oxide synthase, bcl-2 and Fas expression in cultured human coronary endothelial cells exposed to anoxia–reoxygenation and angiotensin II: role of AT1 receptor activation

Dayuan Li, Kathleen Tomson, Baichun Yang, Paulette Mehta, Byron P. Croker and Jawahar L. Mehta*

Departments of Medicine, Physiology, Pediatrics and Pathology, University of Florida and VA Medical Center, 1600 Archer Rd., P.O. Box 100277, JHMHC Gainesville, FL 32610, USA

* Corresponding author. Tel.: +1-(352)-379-4160; fax: +1-(352)-379-4161; e-mail: mehta@medmac.ufl.edu

Background: Angiotensin II (Ang II) plays a critical role in the pathophysiology of myocardial ischemia–reperfusion injury. We have recently shown that reoxygenation following a period of anoxia causes apoptosis of cultured human coronary artery endothelial cells (HCAECs). Ang II further enhances apoptosis of HCAECs via Ang II type 1 receptor (AT1R) activation. Recent studies suggest an important role of constitutive nitric oxide synthase (cNOS), Fas and bcl-2 proteins in apoptosis. This study was designed to examine the modulation of cNOS, and Fas and bcl-2 expression in HCAECs during exposure to anoxia–reoxygenation and Ang II. Methods and Results: HCAECs were exposed to anoxia–reoxygenation and Ang II. Anoxia- reoxygenation significantly decreased cNOS mRNA, protein and activity in cultured HCAECs (P<0.05 vs. control). Anoxia–reoxygenation also caused an increase in Fas and a decrease in bcl-2 protein expression in cultured HCAECs (both P<0.05 vs. control). The presence of Ang II (0.3 µM) further enhanced these effects of anoxia–reoxygenation on cNOS, Fas and bcl-2 expression. The effects of Ang II were significantly attenuated by the AT1R inhibitor losartan (10 µM). Conclusion: During exposure of HCAECs to anoxia–reoxygenation and Ang II, AT1R activation induces important changes in cNOS mRNA, protein expression and activity, as well as bcl-2 and Fas protein expression which may have a bearing on the development of apoptosis.

KEYWORDS Ang II, Angiotensin II; AT1R, Angiotensin II type 1 receptor; AT2R, Angiotensin II type 2 receptor; cNOS, Constitutive nitric oxide synthase; HCAECs, Human coronary artery endothelial cells; iNOS, Inducible nitric oxide synthase


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