Fig 1.
Comparison of coronary reactive hyperemia (CRH) between Tie2-CYP2J2 Tr and WT mice.
Repayment volume (A), repayment duration (B), and repayment/debt ratio (C), were increased in Tie2-CYP2J2 Tr compared to WT mice (P < 0.05). Baseline CF (D) was not different between the two groups. * P < 0.05 versus WT. n = 8 per group.
Fig 2.
LC–MS/MS analysis for EETs’ (8, 9–, 11, 12–, and 14, 15–) and DHETs’ (8, 9–, 11, 12–, and 14, 15–) levels in WT and Tie2-CYP2J2 Tr mouse heart perfusates at baseline and post-ischemia.
11,12-EET (B), and 14,15-EET (C) were increased in Tie2-CYP2J2 Tr compared to WT mice (P < 0.05), whereas 8,9-EET (A) was not significantly different between the two groups (P > 0.05). 8,9-DHET (D), 11,12-DHET (E), and 14,15-DHET (F) were not significantly different between WT and Tie2-CYP2J2 Tr mice. Also, the levels of EETs and DHETs were not affected in response to ischemia in either mouse genotype (P > 0.05). * P < 0.05 versus baseline WT. # P < 0.05 versus WT post-ischemia. n = 12 per group.
Fig 3.
Effect of the CYP-epoxygenases inhibitor, MS-PPOH (1 μM), on coronary reactive hyperemia (CRH) in Tie2-CYP2J2 Tr and WT mice.
The CYP-epoxygenases inhibitor, MS-PPOH, decreased CRH in both Tie2-CYP2J2 Tr and WT mice. (A): MS-PPOH decreased repayment volume in both mouse genotypes. However, repayment volume was still higher in untreated Tie2-CYP2J2 Tr vs. untreated WT, and in MS-PPOH-treated Tie2-CYP2J2 Tr vs. MS-PPOH-treated WT. (B): repayment duration was not changed by the treatment with MS-PPOH in both WT and Tie2-CYP2J2 Tr mice, but repayment duration was increased more in untreated Tie2-CYP2J2 Tr vs. untreated WT. (C): MS-PPOH increased repayment/debt ratio in WT mice, but decreased it in Tie2-CYP2J2 Tr mice. Repayment/debt ratio was increased more in untreated Tie2-CYP2J2 Tr vs. untreated WT, and in MS-PPOH-treated Tie2-CYP2J2 Tr vs. MS-PPOH-treated WT. (D): MS-PPOH decreased baseline CF in both mouse genotypes. There was no difference in baseline CF between untreated Tie2-CYP2J2 Tr and untreated WT. * P < 0.05 versus untreated WT. # P < 0.05 versus untreated Tie2-CYP2J2 Tr mice. Ψ P < 0.05 versus MS-PPOH-treated WT. n = 8 per group.
Fig 4.
LC–MS/MS analysis of 5-, 8-, 11-, 12- and 15-HETE levels in WT and Tie2-CYP2J2 Tr mouse heart perfusates at baseline and post-ischemia.
In Tie2-CYP2J2 Tr mice, the levels of 8-HETE (B) and 12-HETE (D) were increased compared to WT mice at baseline and post-ischemia (P < 0.05). In both WT and Tie2-CYP2J2 Tr mice, post-ischemic levels of 5-HETE (A), 8-HETE (B), 11-HETE (C), 12-HETE (D) and 15-HETE (E) were decreased compared to baseline levels (P < 0.05). * P < 0.05 versus baseline WT. # P < 0.05 versus WT post-ischemia. Ф P < 0.05 versus baseline Tie2-CYP2J2 Tr. n = 12 per group.
Fig 5.
Effect of the CYP4A-blocker, DDMS (1 μM), on coronary reactive hyperemia (CRH) in Tie2-CYP2J2 Tr and WT mice.
The CYP4A-blocker, DDMS, enhanced CRH in both Tie2-CYP2J2 Tr and WT mice. Repayment volume (A) and repayment duration (B) were increased by DDMS in both mouse genotypes. Both repayment volume (A) and repayment duration (B) were increased more in untreated Tie2-CYP2J2 Tr compared to untreated WT mice. Similarly, repayment/debt ratio (C) was increased by DDMS in both mouse genotypes, but was significant in WT mice only. Also, repayment/debt ratio was increased more in untreated Tie2-CYP2J2 Tr compared to untreated WT mice. Baseline CF (D) was not different between the two groups. * P < 0.05 versus untreated WT. # P < 0.05 versus untreated Tie2-CYP2J2 Tr mice. n = 8 per group.
Fig 6.
LC–MS/MS analysis of 6-keto-PG-F1α, PG-F2α, PG-D2, PG-E2, and TxB2 in WT and Tie2-CYP2J2 Tr mouse heart perfusates at baseline and post-ischemia.
The levels of prostaglandin 6-Keto-PG-F1α (A), PG-D2 and (C), and thromboxane (Tx)-B2 (E) were not significantly changed due to ischemia or between Tie2-CYP2J2 Tr and WT mice (P > 0.05). In both Tie2-CYP2J2 Tr and WT mice, PG-F2α (B) and PG-E2 and (D) were decreased post-ischemia (P < 0.05). * P < 0.05 versus baseline WT. Ф P < 0.05 versus baseline Tie2-CYP2J2 Tr. n = 8 per group.
Fig 7.
LC–MS/MS analysis of EpOME and DiHOME levels in WT and Tie2-CYP2J2 Tr mouse heart perfusates at baseline and post-ischemia.
9,10- and 12,13-EpOMEs (A) were increased in Tie2-CYP2J2 Tr compared to WT mice at baseline and post-ischemia (P < 0.05). 9,10- and 12, 13-DiHOMEs (B) did not change between Tie2-CYP2J2 Tr compared to WT mice (P > 0.05). * P < 0.05 versus baseline WT. # P < 0.05 versus WT post-ischemia. n = 8 per group.
Fig 8.
LC–MS/MS analysis of HODEs in WT and Tie2-CYP2J2 Tr mouse heart perfusates at baseline and post-ischemia.
In both WT and Tie2-CYP2J2 Tr, 9- and 13-HODEs decreased in response to ischemia (P < 0.05). However, they were not significantly different between the two groups at baseline or post-ischemia (P > 0.05). * P < 0.05 versus baseline WT. Ф P < 0.05 versus WT post-ischemia. n = 8 per group.
Fig 9.
LC–MS/MS analysis of 8-iso-PGF2α in WT and Tie2-CYP2J2 Tr mouse heart perfusates at baseline and post-ischemia.
8-Iso-PGF2α decreased in Tie2-CYP2J2 Tr compared to WT mice at baseline and post-ischemia (P < 0.05). However, in both mouse genotypes, 8-Iso-PGF2α was not significantly changed in response to ischemia (P > 0.05). *P < 0.05 versus baseline WT. #P < 0.05 versus WT post-ischemia. n = 8 per group.
Fig 10.
A schematic diagram comparing the oxylipin changes observed in response to both brief ischemia and CYP 2J2 over-expression as well as oxylipins’ possible effect on coronary reactive hyperemia (CRH) in WT and Tie2-CYP2J2 Tr mice.
Both ischemia and CYP 2J2 over-expression caused changes in the measured oxylipin profiles. However, the changes associated with ischemia were common to both mouse strains (WT and Tie2-CYP2J2 Tr). Overall, CRH was enhanced in Tie2-CYP2J2 Tr compared to WT possibly through increased EETs, increased EpOMEs, and decreased 8-iso-PGF2α. Grey block arrows indicate the effects of ischemia, whereas black block arrows indicate the effects of ischemia CYP 2J2 over-expression.