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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. 2011 Sep; 28(3):405-12.IJ

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.

Authors+Show Affiliations

Department of Nephrology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, PR China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21537828

Citation

Chang, Jie, et al. "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, vol. 28, no. 3, 2011, pp. 405-12.
Chang J, Jiang Z, Zhang H, et al. 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. 2011;28(3):405-12.
Chang, J., Jiang, Z., Zhang, H., Zhu, H., Zhou, S. F., & 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(3), 405-12. https://doi.org/10.3892/ijmm.2011.683
Chang J, et al. 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. 2011;28(3):405-12. PubMed PMID: 21537828.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - NADPH oxidase-dependent formation of reactive oxygen species contributes to angiotensin II-induced epithelial-mesenchymal transition in rat peritoneal mesothelial cells. AU - Chang,Jie, AU - Jiang,Zongpei, AU - Zhang,Haiyan, AU - Zhu,Hengmei, AU - Zhou,Shu-Feng, AU - Yu,Xueqing, Y1 - 2011/04/28/ PY - 2011/02/15/received PY - 2011/03/31/accepted PY - 2011/5/4/entrez PY - 2011/5/4/pubmed PY - 2011/11/1/medline SP - 405 EP - 12 JF - International journal of molecular medicine JO - Int. J. Mol. Med. VL - 28 IS - 3 N2 - 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. SN - 1791-244X UR - https://www.unboundmedicine.com/medline/citation/21537828/NADPH_oxidase_dependent_formation_of_reactive_oxygen_species_contributes_to_angiotensin_II_induced_epithelial_mesenchymal_transition_in_rat_peritoneal_mesothelial_cells_ L2 - http://www.spandidos-publications.com/ijmm/28/3/405 DB - PRIME DP - Unbound Medicine ER -