Cyp2j5‐gene Deletion Affects on Acetylcholine and Adenosine‐induced Relaxation in Mice: Role of Angiotensin‐II, CYP‐epoxygenase and Omega‐hydroxylase inhibitors

Abstract

Previously, we showed vascular endothelial overexpression of human‐CYP2J2 enhances coronary reactive hyperemia in Tie2‐CYP2J2 Tr mice, but we did not see the response in CYP2J‐epoxygenase knockout mice. Therefore, we hypothesized that Cyp2j5‐gene deletion affects acetylcholine‐, NECA (adenosine)‐and CGS 21680 (adenosine A2A receptor)‐induced relaxation and their response is partially inhibited by angiotensin‐II (Ang‐II) and ω‐hydroxylase inhibitor in mice. ACh‐induced response was tested with N‐(methylsulfonyl)‐2‐(2‐propynyloxy)‐benzenehexanamide (MS‐PPOH, CYP‐epoxygenase inhibitor; 10–5M) and Ang‐II (10‐6M). In Cyp2j5−/− mice, ACh‐induced relaxation was different from C57Bl/6 mice, at 10‐5 M (76.1 ± 3.3 vs. 58.3 ± 5.2, P<0.05). However, ACh‐induced relaxation was not blocked by MS‐PPOH in Cyp2j5−/−: 58.5 ± 5.0%, P>0.05, but blocked in C57Bl/6: 52.3 ± 7.5%, P<0.05, and Ang‐II reduces ACh‐induced relaxation in both Cyp2j5−/− and C57Bl/6 mice (38.8 ± 3.9% and 45.9 ± 7.8, P <0.05). In addition, NECA‐induced response tested with Ang‐II and 20‐HETE inhibitor, dibromo‐dodecenyl‐methylsulfimide (DDMS, 10‐5M). In Cyp2j5−/− mice, NECA‐induced response was not different from C57Bl/6 mice at 10‐5M (23.1 ± 2.1 vs. 21.1 ± 3.8, P>0.05). However, NECA‐induced response was reduced by Ang‐II in both Cyp2j5−/− and C57Bl/6 mice (−10.8 ± 2.3% and 3.2 ± 2.7, P <0.05), and the reduced response due to Ang‐II was partially rescued by DDMS in both Cyp2j5−/− and C57Bl/6 mice (at 10‐5M, A2AAR‐agonist, CGS‐21680, 15.8 ± 3.4% and 13.0 ± 2.3, P <0.05). Data suggest that ACh‐induced relaxation in Cyp2j5−/− mice depends on nitric oxide (NO) but not CYP‐epoxygenases, and the NECA‐induced different response in male vs. female Cyp2j5−/− mice when Ang‐II treated.Support or Funding InformationNational Institutes of Health (HL‐114559) to M. A. Nayeem and the Intramural Research Program of the National Institute of Environmental Health Sciences (Z01 ES025034) to D. C. Zeldin supported this work.Flow diagram: Comparison between Cyp2j5−/− vs. C57Bl/6 mice treated with Acetylcholine, Acetylcholine + MS‐PPOH, Acetylcholine + Ang‐II, NECA, NECA + Ang‐II and CGS 21680 + DDMS + Ang‐II.Figure 1