TY - JOUR
T1 - The protective effect of astrocyte-derived 14,15-epoxyeicosatrienoic acid on hydrogen peroxide-induced cell injury in astrocyte-dopaminergic neuronal cell line co-culture
AU - Terashvili, M.
AU - Sarkar, P.
AU - Nostrand, M. V.
AU - Falck, J R
AU - Harder, D. R.
N1 - Funding Information:
This work was supported by the National Institutes of Health [ P01 HL059996 , R01 HL033833 and R01 HL092105 ], and also by the Veterans’ Association – Dr. Harder is a Research Career Scientist.
PY - 2012/10/25
Y1 - 2012/10/25
N2 - 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 (H2O2). We found that dopaminergic neuronal cells (N27 cell line) stimulated with two different doses of H2O2 (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 H2O2 for 1h. Dopaminergic neuronal cells (N27 cell line) pretreated with different doses of 14,15-EET (0.1-30μM, 30min) before H2O2 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 H2O2 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 (1-20μM, 1h) before H2O2 (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.
AB - 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 (H2O2). We found that dopaminergic neuronal cells (N27 cell line) stimulated with two different doses of H2O2 (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 H2O2 for 1h. Dopaminergic neuronal cells (N27 cell line) pretreated with different doses of 14,15-EET (0.1-30μM, 30min) before H2O2 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 H2O2 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 (1-20μM, 1h) before H2O2 (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.
KW - 14,15-EET
KW - AUDA
KW - HO
KW - Neuron-astrocyte co-culture
KW - Parkinson's disease
KW - Soluble epoxide hydrolase
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U2 - 10.1016/j.neuroscience.2012.07.045
DO - 10.1016/j.neuroscience.2012.07.045
M3 - Article
C2 - 22863680
AN - SCOPUS:84865450193
SN - 0306-4522
VL - 223
SP - 68
EP - 76
JO - Neuroscience
JF - Neuroscience
ER -