20-Hydroxyeicosatetraenoic acid inhibits the apoptotic responses in pulmonary artery smooth muscle cells

• Published in: European journal of pharmacology Vol. 588; no. 1; pp. 9 - 17
• Main Authors: Wang, Zhigang, Tang, Xiaobo, Li, Yumei, Leu, Changlian, Guo, Lei, Zheng, Xiaodong, Zhu, Daling
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 24.06.2008; Elsevier
• Subjects: 20-Hydroxyeicosatetraenoic acid; Annexin A5 - metabolism; Apoptosis; Apoptosis - drug effects; Benzimidazoles; Biological and medical sciences; Blotting, Western; Caspase 3 - metabolism; Caspase 9 - metabolism; Chromatin - chemistry; Coloring Agents; Culture Media, Serum-Free; Cytochrome P450 4A; Fatty Acids, Unsaturated - pharmacology; Hydroxyeicosatetraenoic Acids - pharmacology; In Situ Nick-End Labeling; L-Lactate Dehydrogenase - metabolism; Medical sciences; Membrane Potentials - drug effects; Mitochondrial Membranes - drug effects; Myocytes, Smooth Muscle - drug effects; Pharmacology. Drug treatments; Propidium; Proto-Oncogene Proteins c-bcl-2 - metabolism; Pulmonary Artery - cytology; Pulmonary Artery - drug effects; Pulmonary artery smooth muscle cells; Tetrazolium Salts; Thiazoles; Cytochrome P450 4A; 20-Hydroxyeicosatetraenoic acid; Pulmonary artery smooth muscle cells; Apoptosis; Enzyme; Cytochrome P450; Smooth muscle; In vitro; Pulmonary artery; Pulmonary vessel; Cell death; Blood vessel; Circulatory system
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2008.03.045

20-Hydroxyeicosatetraenoic acid (20-HETE), a ω-hydroxylation product of arachidonic acid catalyzed by cytochrome P450 4A (CYP4A), plays a role in vascular smooth muscle remodeling. Although its effects on angiogenic responses are known, it remains unclear whether 20-HETE acts on apoptosis of pulmonary arterial smooth muscle cells (PASMC), an important step in PASMC remodeling, and what pathways are involved in the process. Here we show evidence for the missing information. The effect of 20-HETE on PASMC apoptosis and the apoptosis-associated signaling pathways were determined with cell viability assay, Annexin V and propidium idodide binding, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL), mitochondrial potentials assay, caspase activity assay and Western blots. We found that exogenous 20-HETE suppressed the serum deprivation-induced loss of bovine PASMCs and prevented Annexin V binding, DNA nick end labeling and chromatin condensation. The effect was worsened by 17-octadecynoic acid (17-ODYA), which inhibited the production of endogenous 20-HETE. Furthermore, 20-HETE induced the expression of bcl-2, maintained the stability of mitochondria membrane, and relieved the activation of caspase-9 and caspase-3. Such effects were reversed in the presence of 17-ODYA. Thus, these findings indicate that 20-HETE protects PASMCs against apoptosis by acting on, at least in part, the intrinsic apoptotic pathway.