Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII
Previous studies have demonstrated the important role of angiotension II (AngII) in promoting proliferation of myofibroblasts (myoFbs) and myocardial fibrosis. However, the underlying mechanisms and the role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII are unclear....
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| Format: | Article |
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Elsevier
2013-02-01
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| Series: | Acta Pharmaceutica Sinica B |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211383512001608 |
| _version_ | 1811223363597631488 |
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| author | Liying Wang Hong Li Shijie Yang |
| author_facet | Liying Wang Hong Li Shijie Yang |
| author_sort | Liying Wang |
| collection | DOAJ |
| description | Previous studies have demonstrated the important role of angiotension II (AngII) in promoting proliferation of myofibroblasts (myoFbs) and myocardial fibrosis. However, the underlying mechanisms and the role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII are unclear. The present study was designed to shed light on this issue through exploration of AngII signaling pathways via in vitro experiments. Primary cultures of neonatal rat myoFbs were divided into five groups which were treated with AngII (10−8 to 10−6 M), AngII with the antioxidant N-acetyl-L-cysteine (NAC), or normal culture medium. We observed the proliferation of myoFbs as induced by AngII at different concentrations with MTT. Reactive oxygen species (ROS) levels in myoFbs were detected by monitoring the fluorescence of 2′,7′-dichlorofluorescein. The contents and levels of oxygen free radicals (OH·) in the three groups were detected by spectrophotometer, immunocytochemical staining, and confocal fluorescence. Western blot and image analysis were used to measure membrane translocation and expression of phospho-protein kinase Cα. MyoFbs incubated with AngII (10−8 to 10−6 M) for 24 h increased their rate of proliferation, the content of OH·, and expression of ROS (P<0.01 vs. control group), whereas these parameters decreased in the presence of NAC. Immunocytochemistry, confocal fluorescence staining and image analysis showed that AngII could promote the translocation and expression of p-PKCα in membrane, and the antioxidant NAC blocked this increase (P<0.01). Western blot results also showed that NAC could inhibit the expression of p-PKCα. |
| first_indexed | 2024-04-12T08:31:40Z |
| format | Article |
| id | doaj.art-1e6a6e0d923a49f4887abb689cb8d7b4 |
| institution | Directory Open Access Journal |
| issn | 2211-3835 |
| language | English |
| last_indexed | 2024-04-12T08:31:40Z |
| publishDate | 2013-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Acta Pharmaceutica Sinica B |
| spelling | doaj.art-1e6a6e0d923a49f4887abb689cb8d7b42022-12-22T03:40:10ZengElsevierActa Pharmaceutica Sinica B2211-38352013-02-0131323710.1016/j.apsb.2012.12.001Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngIILiying Wang0Hong Li1Shijie Yang2Department of Pharmacology, Norman Bethune College of Medicine, Jilin University, Changchun 130021, ChinaDepartment of Pharmacology, Norman Bethune College of Medicine, Jilin University, Changchun 130021, ChinaDepartment of Pharmacology, Norman Bethune College of Medicine, Jilin University, Changchun 130021, ChinaPrevious studies have demonstrated the important role of angiotension II (AngII) in promoting proliferation of myofibroblasts (myoFbs) and myocardial fibrosis. However, the underlying mechanisms and the role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII are unclear. The present study was designed to shed light on this issue through exploration of AngII signaling pathways via in vitro experiments. Primary cultures of neonatal rat myoFbs were divided into five groups which were treated with AngII (10−8 to 10−6 M), AngII with the antioxidant N-acetyl-L-cysteine (NAC), or normal culture medium. We observed the proliferation of myoFbs as induced by AngII at different concentrations with MTT. Reactive oxygen species (ROS) levels in myoFbs were detected by monitoring the fluorescence of 2′,7′-dichlorofluorescein. The contents and levels of oxygen free radicals (OH·) in the three groups were detected by spectrophotometer, immunocytochemical staining, and confocal fluorescence. Western blot and image analysis were used to measure membrane translocation and expression of phospho-protein kinase Cα. MyoFbs incubated with AngII (10−8 to 10−6 M) for 24 h increased their rate of proliferation, the content of OH·, and expression of ROS (P<0.01 vs. control group), whereas these parameters decreased in the presence of NAC. Immunocytochemistry, confocal fluorescence staining and image analysis showed that AngII could promote the translocation and expression of p-PKCα in membrane, and the antioxidant NAC blocked this increase (P<0.01). Western blot results also showed that NAC could inhibit the expression of p-PKCα.http://www.sciencedirect.com/science/article/pii/S2211383512001608N-acetyl-L-cysteineAngIIOxygen free radicalMyofibroblastsProtein kinase Cα |
| spellingShingle | Liying Wang Hong Li Shijie Yang Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII Acta Pharmaceutica Sinica B N-acetyl-L-cysteine AngII Oxygen free radical Myofibroblasts Protein kinase Cα |
| title | Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII |
| title_full | Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII |
| title_fullStr | Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII |
| title_full_unstemmed | Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII |
| title_short | Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII |
| title_sort | role of oxygen free radicals in the proliferation of myofibroblasts induced by angii |
| topic | N-acetyl-L-cysteine AngII Oxygen free radical Myofibroblasts Protein kinase Cα |
| url | http://www.sciencedirect.com/science/article/pii/S2211383512001608 |
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