title: The protective effect of astrocyte-derived 14,15-epoxyeicosatrienoic acid on hydrogen peroxide-induced cell injury in astrocyte-dopaminergic neuronal cell line co-culture.
abstract: Astrocytes perform several functions that are essential for normal neuronal activity. They play a critical role in neuronal survival during ischemia and other degenerative injuries and also modulate neuronal recovery by influencing neurite outgrowth. In this study, we investigated the neuroprotective effects of astrocyte-derived 14,15-epoxyeicosatrienoic acid (14,15-EET), metabolite of arachidonic acid by cytochrome P450 epoxygenases (CYP), against oxidative stress induced by hydrogen peroxide (H(2)O(2)). We found that dopaminergic neuronal cells (N27 cell line) stimulated with two different doses of H(2)O(2) (0.1 and 1mM) for 1h showed decreased cell viability compared to the control group, while astrocytes showed less cell death after stimulation with the same doses of H(2)O(2) for 1h. Dopaminergic neuronal cells (N27 cell line) pretreated with different doses of 14,15-EET (0.1-30 μM, 30 min) before H(2)O(2) stimulation also showed increased cell viability. Furthermore, pre-treatment of the co-cultured cells with 12-(3-adamantan-1-yl-ureido)-dodecanoic acid, an inhibitor of the EET metabolizing enzyme, soluble epoxide hydrolase (sEH), before H(2)O(2) stimulation (1mM, for 1h) increased cell viability. It also increased the endogenous level of 14,15-EET in the media compared to control group. However, pretreatment with the CYP epoxygenase inhibitor miconazole (1-20 μM, 1h) before H(2)O(2) (1mM, 1h) stimulation showed decreased cell viability. Our data suggest that 14,15-EET which is released from astrocytes, enhances cell viability against oxidant-induced injury. Further understanding of the mechanism of 14,15-EET-mediated protection in dopaminergic neurons is imperative, as it could lead to novel therapeutic approaches for treating CNS neuropathologies, such as Parkinson's disease.
authors.affiliation: Department of Physiology, Cardiovascular Research Center, Medical College of Wisconsin, Milwaukee, WI 53226, United States.
authors.forename: M
authors.initials: M
authors.lastname: Terashvili
authors.forename: P
authors.initials: P
authors.lastname: Sarkar
authors.forename: M V
authors.initials: MV
authors.lastname: Nostrand
authors.forename: J R
authors.initials: JR
authors.lastname: Falck
authors.forename: D R
authors.initials: DR
authors.lastname: Harder
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D018920:Coculture Techniques; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004334:Drug Administration Schedule; D015777:Eicosanoids / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D006861:Hydrogen Peroxide / Q000633:toxicity; D013058:Mass Spectrometry; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D058632:Neuroprostanes / Q000494:pharmacology*; D016877:Oxidants / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: D015777:Eicosanoids; D058632:Neuroprostanes; D016877:Oxidants; C046782:14,15-epoxy-5,8,11-eicosatrienoic acid; D006861:Hydrogen Peroxide; D015126:8,11,14-Eicosatrienoic Acid