Abstract 071: Cytokine-angiotensin II Interplay in Cyclophilin D-mediated Vascular Oxidative Stress and Hypertension
Vascular inflammation and oxidative stress interact in a feed-forward fashion to promote vascular disease and hypertension. We hypothesized that angiotensin II and inflammatory cytokines encountered in hypertension co-operatively induce superoxide (O2•-) production by mitochondrial complex I and that efforts to reduce complex I O2•- will reduce hypertension. Treatment of human aortic endothelial cells in culture with angiotensin II (10 nM), IL17A (10 nM) and TNFα (1 nM), factors known to contribute to the hypertensive milieu, co-operatively induced mitochondrial O2•- from 340 to 958 pmol/mg protein as measured by HPLC and MitoSOX. This response was abolished by the complex I inhibitor rotenone. We further tested a potential role of Cyclophilin D (CypD), the redox sensitive regulatory subunit of the mitochondrial transition pore in complex I O2•- production. Both the specific CypD inhibitor Sanglifehrin A and knockdown of CypD by siRNA prevented endothelial cell O2•- production in response to Ang/IL17/TNF. We also found that this cytokine-angiotensin II milieu induced S-glutathionylation of CypD and that scavenging mitochondrial H2O2 with mitoEbselen prevents this and eliminates CypD dependent complex I O2•- production. We further studied the functional role of oxidative stress induced by Ang/IL17/TNF in isometric tension studies of mouse aortic rings. Twenty-four hour treatment of organoid cultured vessels with AngII/IL17/TNF reduced endothelium-dependent vasodilatation to acetylcholine and this was prevented by knockdown of CypD and was not observed in vessels of mice with overexpression of mitochondrial SOD or mitochondrial catalase. The in vivo role of CypD in regulation of vascular O2•- and blood pressure was further studied in mice infused with angiotensin II (490 ng/kg/min). Treatment with Sanglifehrin A after the onset of hypertension reduced blood pressure from 162 to 133 mmHg (P<0.01), reduced vascular O2•- and improved endothelium-dependent vasodilation. These studies have defined a novel role of Cyclophilin D as a cause of vascular dysfunction and hypertension and have provided a new target for treatment of this disease.
Author Disclosures: R. Nazarewicz: None. A. Dikalova: None. H. Itani: None. W. McMaster: None. A. Bikineyeva: None. D.G. Harrison: None. S. Dikalov: None.
This research has received full or partial funding support from the American Heart Association, Greater Southeast Affiliate (Alabama, Florida, Georgia, Louisiana, Mississippi, Puerto Rico & Tennessee).
- © 2014 by American Heart Association, Inc.