Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth
Reactive oxygen species (ROS) including nitric oxide (NO) and superoxide anion (O2-) are associated with cell migration, proliferation and many growth-related diseases. The objective of this study was to determine whether there was a reciprocal relationship between rat coronary microvascular endothe...
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| Format: | Article |
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2004
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| Online Access: | https://eprints.nottingham.ac.uk/486/ |
| _version_ | 1848790418672582656 |
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| author | Bayraktutan, Ulvi |
| author_facet | Bayraktutan, Ulvi |
| author_sort | Bayraktutan, Ulvi |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Reactive oxygen species (ROS) including nitric oxide (NO) and superoxide anion (O2-) are associated with cell migration, proliferation and many growth-related diseases. The objective of this study was to determine whether there was a reciprocal relationship between rat coronary microvascular endothelial cell (CMEC) growth and activity/expressions (mRNA and protein) of endothelial NO synthase (eNOS) and NAD(P)H oxidase enzymes. Proliferating namely, 50% confluent CMEC possessed approximately three-fold increased activity and expression of both enzymes compared to 100% confluent cells. Treatment of CMEC with an inhibitor of eNOS (L-NAME, 100M) increased cell proliferation as assessed via three independent methods i.e. cell counting, determination of total cellular protein levels and [3H]thymidine incorporation.
Similarly, treatment of CMEC with pyrogallol (0.3-3 mM), a superoxide anion (O2-)- generator, also increased CMEC growth while spermine NONOate (SpNO), a NO donor, significantly reduced cell growth. Co-incubation of CMEC with a cell permeable superoxide dismutase mimetic (Mn-III-tetrakis-4-benzoic acid-porphyrin; MnTBAP) plus either pyrogallol or NO did not alter cell number and DNA synthesis thereby dismissing the involvement of peroxynitrite (OONO-) in CMEC proliferation. Specific inhibitors of NAD(P)H oxidase but not other ROS-generating enzymes including cyclooxygenase and xanthine oxidase, attenuated cell growth. Transfection of CMEC with antisense p22-phox cDNA, a membrane-bound component of NAD(P)H oxidase, resulted in substantial reduction in [3H]thymidine incorporation, total cellular protein levels and expression of p22-phox protein. These data demonstrate a cross-talk between CMEC growth and eNOS and NAD(P)H oxidase enzyme activity and expression, thus suggesting that the regulation of these enzymes may be critical in preventing the initiation and/or progression of coronary atherosclerosis. |
| first_indexed | 2025-11-14T18:12:18Z |
| format | Article |
| id | nottingham-486 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T18:12:18Z |
| publishDate | 2004 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-4862020-05-04T20:31:20Z https://eprints.nottingham.ac.uk/486/ Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth Bayraktutan, Ulvi Reactive oxygen species (ROS) including nitric oxide (NO) and superoxide anion (O2-) are associated with cell migration, proliferation and many growth-related diseases. The objective of this study was to determine whether there was a reciprocal relationship between rat coronary microvascular endothelial cell (CMEC) growth and activity/expressions (mRNA and protein) of endothelial NO synthase (eNOS) and NAD(P)H oxidase enzymes. Proliferating namely, 50% confluent CMEC possessed approximately three-fold increased activity and expression of both enzymes compared to 100% confluent cells. Treatment of CMEC with an inhibitor of eNOS (L-NAME, 100M) increased cell proliferation as assessed via three independent methods i.e. cell counting, determination of total cellular protein levels and [3H]thymidine incorporation. Similarly, treatment of CMEC with pyrogallol (0.3-3 mM), a superoxide anion (O2-)- generator, also increased CMEC growth while spermine NONOate (SpNO), a NO donor, significantly reduced cell growth. Co-incubation of CMEC with a cell permeable superoxide dismutase mimetic (Mn-III-tetrakis-4-benzoic acid-porphyrin; MnTBAP) plus either pyrogallol or NO did not alter cell number and DNA synthesis thereby dismissing the involvement of peroxynitrite (OONO-) in CMEC proliferation. Specific inhibitors of NAD(P)H oxidase but not other ROS-generating enzymes including cyclooxygenase and xanthine oxidase, attenuated cell growth. Transfection of CMEC with antisense p22-phox cDNA, a membrane-bound component of NAD(P)H oxidase, resulted in substantial reduction in [3H]thymidine incorporation, total cellular protein levels and expression of p22-phox protein. These data demonstrate a cross-talk between CMEC growth and eNOS and NAD(P)H oxidase enzyme activity and expression, thus suggesting that the regulation of these enzymes may be critical in preventing the initiation and/or progression of coronary atherosclerosis. 2004 Article PeerReviewed Bayraktutan, Ulvi (2004) Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth. Journal of Molecular and Cellular Cardiology, 36 . pp. 277-286. |
| spellingShingle | Bayraktutan, Ulvi Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth |
| title | Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth |
| title_full | Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth |
| title_fullStr | Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth |
| title_full_unstemmed | Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth |
| title_short | Nitric oxide synthase and NAD(P)H oxidase modulate coronary endothelial cell growth |
| title_sort | nitric oxide synthase and nad(p)h oxidase modulate coronary endothelial cell growth |
| url | https://eprints.nottingham.ac.uk/486/ |