Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats
Kinin B1 receptor (B1R) enhanced superoxide anion (O2•-) production in the vasculature of diabetic rats. This study investigates the induction and distribution of B1R in diabetic blood vessels and addresses the hypothesis that B1R is co-localized with NADPH oxidase (NOX1 and NOX2) and produces its a...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-10-01
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| Series: | Frontiers in Physiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fphys.2017.00861/full |
| _version_ | 1818966661425790976 |
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| author | Youssef Haddad Réjean Couture |
| author_facet | Youssef Haddad Réjean Couture |
| author_sort | Youssef Haddad |
| collection | DOAJ |
| description | Kinin B1 receptor (B1R) enhanced superoxide anion (O2•-) production in the vasculature of diabetic rats. This study investigates the induction and distribution of B1R in diabetic blood vessels and addresses the hypothesis that B1R is co-localized with NADPH oxidase (NOX1 and NOX2) and produces its activation via protein kinase C (PKC). Diabetes was induced in rats with streptozotocin (STZ 65 mg.kg−1, i.p.). Two weeks later, the production of O2•- was measured in aorta rings in response to the B1R agonist (Sar[D-Phe8]-des-Arg9-BK, 20 μM) by the method of lucigenin-enhanced chemiluminescence. Various inhibitors were added (10 μM) to block PKCtotal (Ro-31-8220), PKCβ1/2 (LY333531), or NADPH oxidase (Diphenyleneiodonium). The cellular localization of B1R was studied in the aorta, popliteal artery, and renal glomerulus/arteries by immunofluorescence and confocal microscopy with markers of endothelial cells (anti-RECA-1), macrophages (anti-CD11), vascular smooth muscle cells (anti-SMA), and NADPH oxidase (anti-NOX1 and NOX2). Although B1R was largely distributed in resistant vessels, it was sparsely expressed in the aorta's endothelium. The greater basal production of O2•- in STZ-diabetic aorta was significantly enhanced by the B1R agonist (15–45 min). The peak response to the agonist (30 min) was inhibited by all inhibitors. Immunofluorescent staining for B1R, NOX1, and NOX2 was significantly increased in endothelial cells, vascular smooth muscle cells, and macrophages of STZ-diabetic aorta on which they were found co-localized. Data showed that B1R enhanced O2•- by activating vascular NADPH oxidase through PKCβ1/2. This was substantiated by the cellular co-localization of B1R with NOX1 and NOX2 and opens the possibility that B1R-enhanced oxidative stress is derived from vascular and infiltrating immune cells in diabetes. |
| first_indexed | 2024-12-20T13:36:27Z |
| format | Article |
| id | doaj.art-60e4c0d16bd44332805bcb91b4088f46 |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-20T13:36:27Z |
| publishDate | 2017-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-60e4c0d16bd44332805bcb91b4088f462022-12-21T19:38:55ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2017-10-01810.3389/fphys.2017.00861295092Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic RatsYoussef HaddadRéjean CoutureKinin B1 receptor (B1R) enhanced superoxide anion (O2•-) production in the vasculature of diabetic rats. This study investigates the induction and distribution of B1R in diabetic blood vessels and addresses the hypothesis that B1R is co-localized with NADPH oxidase (NOX1 and NOX2) and produces its activation via protein kinase C (PKC). Diabetes was induced in rats with streptozotocin (STZ 65 mg.kg−1, i.p.). Two weeks later, the production of O2•- was measured in aorta rings in response to the B1R agonist (Sar[D-Phe8]-des-Arg9-BK, 20 μM) by the method of lucigenin-enhanced chemiluminescence. Various inhibitors were added (10 μM) to block PKCtotal (Ro-31-8220), PKCβ1/2 (LY333531), or NADPH oxidase (Diphenyleneiodonium). The cellular localization of B1R was studied in the aorta, popliteal artery, and renal glomerulus/arteries by immunofluorescence and confocal microscopy with markers of endothelial cells (anti-RECA-1), macrophages (anti-CD11), vascular smooth muscle cells (anti-SMA), and NADPH oxidase (anti-NOX1 and NOX2). Although B1R was largely distributed in resistant vessels, it was sparsely expressed in the aorta's endothelium. The greater basal production of O2•- in STZ-diabetic aorta was significantly enhanced by the B1R agonist (15–45 min). The peak response to the agonist (30 min) was inhibited by all inhibitors. Immunofluorescent staining for B1R, NOX1, and NOX2 was significantly increased in endothelial cells, vascular smooth muscle cells, and macrophages of STZ-diabetic aorta on which they were found co-localized. Data showed that B1R enhanced O2•- by activating vascular NADPH oxidase through PKCβ1/2. This was substantiated by the cellular co-localization of B1R with NOX1 and NOX2 and opens the possibility that B1R-enhanced oxidative stress is derived from vascular and infiltrating immune cells in diabetes.http://journal.frontiersin.org/article/10.3389/fphys.2017.00861/fullbradykinin B1 receptorendotheliummacrophagesNOX1NOX2diabetes mellitus |
| spellingShingle | Youssef Haddad Réjean Couture Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats Frontiers in Physiology bradykinin B1 receptor endothelium macrophages NOX1 NOX2 diabetes mellitus |
| title | Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats |
| title_full | Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats |
| title_fullStr | Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats |
| title_full_unstemmed | Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats |
| title_short | Localization and Interaction between Kinin B1 Receptor and NADPH Oxidase in the Vascular System of Diabetic Rats |
| title_sort | localization and interaction between kinin b1 receptor and nadph oxidase in the vascular system of diabetic rats |
| topic | bradykinin B1 receptor endothelium macrophages NOX1 NOX2 diabetes mellitus |
| url | http://journal.frontiersin.org/article/10.3389/fphys.2017.00861/full |
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