NADPH oxidase-dependent formation of reactive oxygen species contributes to angiotensin II-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells

  • Authors:
    • Jie Chang
    • Zongpei Jiang
    • Haiyan Zhang
    • Hengmei Zhu
    • Shu-Feng Zhou
    • Xueqing Yu
  • View Affiliations

  • Published online on: April 28, 2011     https://doi.org/10.3892/ijmm.2011.683
  • Pages: 405-412
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Abstract

The objective of the present study was to investigate the role of NADPH oxidase-dependent formation of reactive oxygen species (ROS) in the angiotensin II (Ang II)-induced epithelial-mesenchymal transition (EMT) and in the accumulation of extracellular matrix (ECM) in rat peritoneal mesothelial cells (RPMCs). Primary cultured RPMCs were incubated with serum-free media for 24 h in order to arrest and synchronize cell growth. The cells were treated with Ang II (10-7 M) up to 48 h. Cells were pretreated with an Ang II type I receptor antagonist (losartan, 10-5 M), or an inhibitor of NADPH, oxidase diphenyleneiodonium (DPI) (10-5 M), for 1 h before addition of Ang II. The dichlorofluorescein (DCF)-sensitive cellular ROS were measured by fluorometric assay and confocal microscopy. RT-PCR was employed to detect the mRNA expression for the NADPH oxidase subunit p47phox, plasminogen activator inhibitor-1 (PAI-1), α-smooth muscle actin (α-SMA) and E-cadherin. PAI-1, α-SMA and p47phox protein expression were examined by Western blot analysis. Ang II significantly induced the production of intracellular ROS. DPI and losartan inhibited Ang II-induced ROS generation by 86.8% and 77.4% (p<0.05), respectively. Ang II significantly stimulated NADPH oxidase subunit p47phox mRNA and protein expression in RPMCs. Both losartan and DPI inhibited Ang II-induced up-regulation of p47phox mRNA by 37.3% and 67.8% (p<0.05), respectively. Ang II also stimulated α-SMA mRNA and protein expression and down-regulated E-cadherin mRNA expression in RPMCs. This effect was suppressed by both losartan and DPI. Ang II significantly up-regulated PAI-1 mRNA and protein expression and these were significantly suppressed by both losartan and DPI. In conclusion, NADPH oxidase-dependent formation of ROS mediates Ang II dependent EMT and accumulation of ECM in rat peritoneal mesothelial cells. NADPH oxidase may represent a potential therapeutic target in the management of peritoneal fibrosis.

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September 2011
Volume 28 Issue 3

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Chang J, Jiang Z, Zhang H, Zhu H, Zhou S and Yu X: NADPH oxidase-dependent formation of reactive oxygen species contributes to angiotensin II-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells. Int J Mol Med 28: 405-412, 2011.
APA
Chang, J., Jiang, Z., Zhang, H., Zhu, H., Zhou, S., & Yu, X. (2011). NADPH oxidase-dependent formation of reactive oxygen species contributes to angiotensin II-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells. International Journal of Molecular Medicine, 28, 405-412. https://doi.org/10.3892/ijmm.2011.683
MLA
Chang, J., Jiang, Z., Zhang, H., Zhu, H., Zhou, S., Yu, X."NADPH oxidase-dependent formation of reactive oxygen species contributes to angiotensin II-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells". International Journal of Molecular Medicine 28.3 (2011): 405-412.
Chicago
Chang, J., Jiang, Z., Zhang, H., Zhu, H., Zhou, S., Yu, X."NADPH oxidase-dependent formation of reactive oxygen species contributes to angiotensin II-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells". International Journal of Molecular Medicine 28, no. 3 (2011): 405-412. https://doi.org/10.3892/ijmm.2011.683