期刊
FREE RADICAL RESEARCH
卷 47, 期 2, 页码 116-122出版社
TAYLOR & FRANCIS LTD
DOI: 10.3109/10715762.2012.749987
关键词
oxidative stress; ACE; cAMP; hypertension; HUVECs
资金
- National Nature Science Foundation of China [30971252, 81170219]
- Funds for Creative Research Groups of the National Natural Science Foundation of China [81121003]
- Major Program of National Natural Science Foundation of China [81130088]
Oxidative stress has been linked to endothelial dysfunction in atherosclerosis and hypertension. The present study was designed to investigate the effect of hydrogen peroxide (H2O2) on angiotensin-converting enzyme (ACE), a key regulator of the renin-angiotensin system, and the mechanisms underlying ACE regulation in human umbilical vein endothelial cells (HUVECs). We used Tetrazolium bromide (MTT) assay for cell viability, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for cell apoptosis, enzyme-linked immunosorbent assay (ELISA) for cAMP measurement, real-time PCR for mRNA detection, and Western blot for protein analysis in the study. Our results demonstrated that H2O2 (50-1000 mu M) decreased HUVECs viability by inducing apoptosis. Notably, H O-2(2) upregulated ACE expression in a concentration-dependent manner. H2O2 100 mu M significantly enhanced cyclic adenosine monophosphate (cAMP) expression by 1.48-fold (P < 0.05). Additionally, forskolin 10 mu M, a cAMP agonist, was also found to enhance ACE expression by 1.78-fold (P < 0.05); in contrast, H-89 10 mu M, a protein kinase A (PKA) inhibitor, abolished H2O2-induced ACE expression and prevented the enhancing effect of forskolin-induced ACE expression. Similar effects on ACE mRNA were also observed. cAMP-response element-specific decoy oligodeoxynucleotides (CRE-dODN) containing binding sites for cAMP-response element-binding protein (CREB) inhibited ACE expression at both the mRNA and protein levels. Negative control CRE-dODN had no effect on ACE expression. We conclude that H2O2 upregulates the expression of ACE through the activation of cAMP/PKA/CREB signal pathway in HUVECs, indicating a role of oxidative stress in the pathophysiology of hypertension.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据