期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 45, 期 9, 页码 1223-1231出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2008.06.012
关键词
Hypoxia; NADPH oxidase; Reactive oxygen species; Intracellular calcium; Mitochondria; Protein kinase C; Pulmonary artery; Free radicals
资金
- AHA Scientist Development [0630236N, 0730242N]
- EHS Center [P30ES06639]
- AHA Established Investigator Award [034016ON]
- NIH [R01HL064043, R01HL075190]
The importance of NADPH oxidase (Nox) in hypoxic responses in hypoxia-sensing cells, including pulmonary artery smooth muscle cells (PASMCs), remains uncertain. In this study, using Western blot analysis we found that the major Nox subunits Nox1, Nox4, p22(phox), p47(phox), and p67(phox) were equivalently expressed in mouse pulmonary and systemic (mesenteric) arteries. However, acute hypoxia significantly increased Nox activity and translocation of p47(phox),, protein to the plasma membrane in pulmonary, but not mesenteric, arteries. The Nox inhibitor apocynin and p47(phox) gene deletion attenuated the hypoxic increase in intracellular concentrations of reactive oxygen species and Ca2+ ([ROS](i) and [Ca2+](i)), as well as contractions in mouse PASMCs, and abolished the hypoxic activation of Nox in pulmonary arteries. The conventional/novel protein kinase C (PKC) inhibitor chelerythrine, specific PKC epsilon translocation peptide inhibitor, and PKC epsilon gene deletion, but not the conventional PKC inhibitor GO6976, prevented the hypoxic increase in Nox activity in pulmonary arteries and [ROS](i) in PASMCs. The PKC activator phorbol 12-myristate 13-acetate could increase Nox activity in pulmonary and mesenteric arteries. Inhibition of mitochondrial ROS generation with rotenone or myxothiazol prevented hypoxic activation of Nox. Glutathione peroxidase-1 (Gpx1) gene overexpression to enhance H2O2 removal significantly inhibited the hypoxic activation of Nox, whereas Gpx1 gene deletion had the opposite effect. Exogenous H2O2 increased Nox activity in pulmonary and mesenteric arteries. These findings suggest that acute hypoxia may distinctively activate Nox to increase [ROS](i) through the mitochondrial ROS-PKC epsilon signaling axis, providing a positive feedback mechanism to contribute to the hypoxic increase in [ROS](i) and [Ca2+](i) as well as contraction in PASMCs. (c) 2008 Elsevier Inc. All rights reserved.
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