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《Chinese Pharmacological Bulletin》 2007-12
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Neuroprotective effects of acetylcholine-mediated EDHF in primarily cultured hippocamal neurons insulted by hypoxia-reoxygenation

WU Jian,CHEN Zhi-wu(Dept of Pharmacology,Anhui Medical University,Hefei 230032,China)  
Aim To study the protective effects of Presumptive endotheliumartmendependent hyperpolarizing factor(EDHF)released from the rat middle cerebral arteries(MCA),which was mediated by acetylcholine(ACh),on primarily cultured hippocampal neurons subjected to hypoxia-reoxygenation injury.Methods Primarily cultured hippocampal neurons was insulted by hypoxia-reoxygenation;EDHF was produced in rat MCA ring by 1 μmol·L-1 ACh in the presence of NG-nitro-L-argininemethyl ester(L-NAME,a NOS inhibitor)and indomethcacin(Indo,a COX inhibitor);MTT absorbance and the LDH activity were served as the cell injury index,The level of free calcium fluorescence intensity in the cultured hippocampal neurons was monitored by laser scanning confocal microsope.Results Compared with nomal group,MTT absorbance were decreased significantly,LDH activity in the supernate culture fluid and the Ca2+ in hippocampal neurons increased significantly in model group.The conjoined use of 1 μmol·L-1 ACh and the endothelium-intacted MCA(MCA/Endo)or ACh+MCA/Endo+L-NAME+Indo can repress both the decrease of MTT absorbance and the increases of LDH activity in the supernate culture fluid and the Ca2+ in neurons which resulted from hypoxia-reoxygenation injury;Neither 1 μmol·L-1ACh alone nor MCA/Endo alone has the similar effects mentioned above,the conjoined use of 1 μmol·L-1 ACh and the endothelium-denuded MCA(ACh+MCA/-Endo)also has little effect.K+ which concentration is between 25~35 μmol·L-1 can significantly attenuate the effects afforded by the conjoined use of 1 μmol·L-1 ACh,MCA/Endo,L-NAME and Indo,but it is not the same situation when it comes to Ba2+.Conclusions Presumptive EDHF can protect primarily cultured rat hippocamal neurons insulted by hypoxia-reoxygenation and the mechanism is partially concerned with the inhibition of calcium overload.
【Fund】: 安徽省人才专项基金资助项目(No2005Z.031)
【CateGory Index】: R363
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