TY - JOUR
T1 - Novel mechanisms of myocardial ischemia, ischemia-reperfusion, and protection by myocardial conditioning
T2 - Infarct size-limiting effect of epoxyeicosatrienoic acid analog EET-B is mediated by hypoxia-inducible factor-1α via downregulation of prolyl hydroxylase 3
AU - Neckář, Jan
AU - Hsu, Anna
AU - Hye Khan, Md Abdul
AU - Gross, Garrett J.
AU - Nithipatikom, Kasem
AU - Cyprová, Michaela
AU - Benák, Daniel
AU - Hlaváčková, Markéta
AU - Sotáková-Kašparová, Dita
AU - Falck, J R
AU - Sedmera, David
AU - Kolář, František
AU - Imig, John D.
N1 - Funding Information:
J. Neckárˇ was supported by Czech Science Foundation Grants 15-07544S and 18-03207S and Institutional Research Projects 67985823 (Institute of Physiology of the Czech Academy of Sciences) and 00023001 (Institute for Clinical and Experimental Medicine). This work was also supported by National Institutes of Health Grants HL-111392 (to G. Gross) and DK-103616 (to J. Imig), a Dr. Ralph and Marian Falk Medical Research Trust Bank of America, North America, Trustee Grant (to J. Imig), and Robert A. Welch Foundation Grant I-0011 (to J. Falck).
Funding Information:
J. Neckář was supported by Czech Science Foundation Grants 15-07544S and 18-03207S and Institutional Research Projects 67985823 (Institute of Physiology of the Czech Academy of Sciences) and 00023001 (Institute for Clinical and Experimental Medicine). This work was also supported by National Institutes of Health Grants HL-111392 (to G. Gross) and DK-103616 (to J. Imig), a Dr. Ralph and Marian Falk Medical Research Trust Bank of America, North America, Trustee Grant (to J. Imig), and Robert A. Welch Foundation Grant I-0011 (to J. Falck).
Publisher Copyright:
© 2018 American Physiological Society. All rights reserved.
PY - 2018/10
Y1 - 2018/10
N2 - Epoxyeicosatr-ienoic acids (EETs) decrease cardiac ischemia-reperfusion injury; however, the mechanism of their protective effect remains elusive. Here, we investigated the cardioprotective action of a novel EET analog, EET-B, in reperfusion and the role of hypoxia-inducible factor (HIF)-1β in such action of EET-B. Adult male rats were subjected to 30 min of left coronary artery occlusion followed by 2h of reperfusion. Administration of 14,15-EET (2.5 mg/kg) or EET-B (2.5 mg/kg) 5 min before reperfusion reduced infarct size expressed as a percentage of the area at risk from 64.3 ± 1.3% in control to 42.6 ± 1.9% and 46.0 ± 1.6%, respectively, and their coadministration did not provide any stronger effect. The 14,15-EET antagonist 14,15-epoxyei-cosa-5(Z)-enoic acid (2.5 mg/kg) inhibited the infarct size-limiting effect of EET-B (62.5 ± 1.1%). Similarly, the HIF-1β inhibitors 2-methoxyestradiol (2.5 mg/kg) and acriflavine (2 mg/kg) completely abolished the cardioprotective effect of EET-B. In a separate set of experiments, the immunoreactivity of HIF-1β and its degrading enzyme prolyl hydroxylase domain protein 3 (PHD3) were analyzed in the ischemic areas and nonischemic septa. At the end of ischemia, the HIF-1β immunogenic signal markedly increased in the ischemic area compared with the septum (10.31 ± 0.78% vs. 0.34 ± 0.08%). After 20 min and 2 h of reperfusion, HIF-1β immunoreactivity decreased to 2.40 ± 0.48% and 1.85 ± 0.43%, respectively, in the controls. EET-B blunted the decrease of HIF-1β immunoreactivity (7.80 ± 0.69% and 6.44 ± 1.37%, respectively) and significantly reduced PHD3 immunogenic signal in ischemic tissue after reperfusion. In conclusion, EET-B provides an infarct size-limiting effect at reperfusion that is mediated by HIF-1β and downregulation of its degrading enzyme PHD3. NEW & NOTEWORTHY The present study shows that EET-B is an effective agonistic 14,15-epoxyeicosatrienoic acid analog, and its administration before reperfusion markedly reduced myocardial infarction in rats. Most importantly, we demonstrate that increased hypoxia-inducible factor-1β levels play a role in cardioprotection mediated by EET-B in reperfusion likely by mechanisms including downregulation of the hypoxia-inducible factor-1β-degrading enzyme prolyl hydroxylase domain protein 3.
AB - Epoxyeicosatr-ienoic acids (EETs) decrease cardiac ischemia-reperfusion injury; however, the mechanism of their protective effect remains elusive. Here, we investigated the cardioprotective action of a novel EET analog, EET-B, in reperfusion and the role of hypoxia-inducible factor (HIF)-1β in such action of EET-B. Adult male rats were subjected to 30 min of left coronary artery occlusion followed by 2h of reperfusion. Administration of 14,15-EET (2.5 mg/kg) or EET-B (2.5 mg/kg) 5 min before reperfusion reduced infarct size expressed as a percentage of the area at risk from 64.3 ± 1.3% in control to 42.6 ± 1.9% and 46.0 ± 1.6%, respectively, and their coadministration did not provide any stronger effect. The 14,15-EET antagonist 14,15-epoxyei-cosa-5(Z)-enoic acid (2.5 mg/kg) inhibited the infarct size-limiting effect of EET-B (62.5 ± 1.1%). Similarly, the HIF-1β inhibitors 2-methoxyestradiol (2.5 mg/kg) and acriflavine (2 mg/kg) completely abolished the cardioprotective effect of EET-B. In a separate set of experiments, the immunoreactivity of HIF-1β and its degrading enzyme prolyl hydroxylase domain protein 3 (PHD3) were analyzed in the ischemic areas and nonischemic septa. At the end of ischemia, the HIF-1β immunogenic signal markedly increased in the ischemic area compared with the septum (10.31 ± 0.78% vs. 0.34 ± 0.08%). After 20 min and 2 h of reperfusion, HIF-1β immunoreactivity decreased to 2.40 ± 0.48% and 1.85 ± 0.43%, respectively, in the controls. EET-B blunted the decrease of HIF-1β immunoreactivity (7.80 ± 0.69% and 6.44 ± 1.37%, respectively) and significantly reduced PHD3 immunogenic signal in ischemic tissue after reperfusion. In conclusion, EET-B provides an infarct size-limiting effect at reperfusion that is mediated by HIF-1β and downregulation of its degrading enzyme PHD3. NEW & NOTEWORTHY The present study shows that EET-B is an effective agonistic 14,15-epoxyeicosatrienoic acid analog, and its administration before reperfusion markedly reduced myocardial infarction in rats. Most importantly, we demonstrate that increased hypoxia-inducible factor-1β levels play a role in cardioprotection mediated by EET-B in reperfusion likely by mechanisms including downregulation of the hypoxia-inducible factor-1β-degrading enzyme prolyl hydroxylase domain protein 3.
KW - Epoxyeicosatrienoic acid
KW - Heart
KW - Hypoxia-inducible factor-1β
KW - Ischemia-reperfusion
KW - Prolyl hydroxylase 3
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U2 - 10.1152/ajpheart.00726.2017
DO - 10.1152/ajpheart.00726.2017
M3 - Article
C2 - 30074840
AN - SCOPUS:85054889895
SN - 0363-6135
VL - 315
SP - H1148-H1158
JO - American Journal of Physiology - Heart and Circulatory Physiology
JF - American Journal of Physiology - Heart and Circulatory Physiology
IS - 5
ER -