20-Hydroxyeicosatetraenoic acid induces apoptosis in neonatal rat cardiomyocytes through mitochondrial-dependent pathways.
OBJECTIVE
20-Hydroxyeicosatetraenoic acid (20-HETE), a [omega]-hydroxylation product of arachidonic acid catalyzed by cytochrome P450 4A, may play a role in the cardiovascular system. It is well known that cytochrome P450 [omega]-hydroxylase inhibitors markedly reduced the cardiac ischemia reperfusion injury. However, the direct effect of 20-HETE on cardiomyocytes is still poorly investigated. Here, we studied the effect of 20-HETE on cardiomyocyte apoptosis and the apoptosis-associated signaling pathways.


RESULTS
The cardiomyocyte apoptosis was measured by fluorescein isothiocyanate conjugated annexin V/propidium iodide double staining cytometry, indicating that the percentage of early apoptotic cells increased from 15.6% +/- 2.6% to 25.5% +/- 2.5% in control and 20-HETE-treated cells, respectively. The mitochondrial membrane potential ([DELTA][PSI]m) was measured by detecting the ratio of JC-1 green/red emission intensity. A significant decrease in the ratio was observed after treatment with 20-HETE for 24 hours in comparison with control group, suggesting the disruptive effect of 20-HETE on mitochondrial [DELTA][PSI]m. In addition, 20-HETE stimulated caspase-3 activity and Bax mRNA expression in cardiomyocytes. In contrast, the Bcl-2 mRNA levels were significantly decreased by 20-HETE treatment.


CONCLUSIONS
These results demonstrate that 20-HETE induces cardiomyocyte apoptosis by activation of several intrinsic apoptotic pathways. The 20-HETE-induced apoptosis could contribute to the cytochrome P450 [omega]-hydroxylase-dependent cardiac injure during cardiac ischemia-reperfusion.
Laboratory of Molecular & Cellular Physiology, School of Life Sciences, Northeast Normal University, Changchun, Jilin, China.
Yuyan
Y
Bao
Xueying
X
Wang
Wei
W
Li
Dan
D
Huo
Xin
X
Shen
Yong
Y
Han
Jiang
J
Tan
Qinghua
Q
Zeng
Chengwen
C
Sun
D000578:Amidines / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D016718:Arachidonic Acid / Q000378:metabolism; D001562:Benzimidazoles; D002232:Carbocyanines; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D042926:Cytochrome P-450 CYP4A / Q000378:metabolism; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000494:pharmacology; D005456:Fluorescent Dyes; D019254:Genes, bcl-2 / Q000187:drug effects; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology*; D006900:Hydroxylation / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects*; D032383:Myocytes, Cardiac / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D013194:Staining and Labeling; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis / Q000187:drug effects
D000578:Amidines; D001562:Benzimidazoles; D002232:Carbocyanines; D065607:Cytochrome P-450 Enzyme Inhibitors; D005456:Fluorescent Dyes; C000708209:HET0016; D006893:Hydroxyeicosatetraenoic Acids; D051028:bcl-2-Associated X Protein; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; D016718:Arachidonic Acid; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D003577:Cytochrome P-450 Enzyme System; D042926:Cytochrome P-450 CYP4A; C409986:cytochrome P-450 omega-hydroxylase; D053148:Caspase 3
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