17β-estradiol通过促进PSD95/nNOS耦联抑制皮层海马神经元的存活

17β-estradiol inhibits the survival of cortical hippocampal neuron by promoting the coupling of nNOS and PSD95

目的:检测17β-estradiol对皮层海马神经元存活的影响,并判断其与nNOS/PSD95耦联的关系。方法:采用原代培养的ICR小鼠皮层海马神经元,培养7天后分溶剂对照组和给药组分别给予溶剂DMSO和浓度为10μmol/L的17β-estradiol,LDH法判断细胞损伤,测定给药30 min后细胞死亡情况,并拍摄光镜照片观察细胞形态。同时利用免疫共沉淀法检测DMSO组,10nmol/L17β-estradiol组,10μmol/L 17β-estradiol组的nNOS和PSD95的耦联情况,观察药物对其影响。结果:给药30 min后,与对照组相比,10μmol/L17β-estradiol组LDH漏出率增加,同时nNOS/PSD95耦联增加,而10 nmol/L 17β-estradiol对nNOS/PSD95耦联并没有影响。结论:浓度为10μmol/L的17β-estradiol会损伤皮层海马神经元,而且这一结果可能是通过促进nNOS/PSD95耦联实现的。

Objective：To detect the effects of 17β-estradiol on the survival of cortical hippocampal neurons and determine whether it is associated with nNOS/PSD95 complexes.Methods：ICR mice was adopted to culture cortical hippocampal neurons.Eight days later,lactate dehydrogenase（LDH） release rate were examined in cultured cortical hippocampal neurons exposed to DMSO or 10 μmol/L 17β-estradiol for 30 min.The neurons were photographed through a light microscope.Meanwhile,nNOS/PSD95 complexes of neurons treated with DMSO,10 nmol/L 17β-estradiol and 10 μmol/L 17β-estradiol were measured by co-immunoprecipitation.Results：At 30 minutes after administration,10 μmol/L 17β-estradiol treatment increased LDH release rate and nNOS/PSD95 conjugating in cortical hippocampal neurons,and 10 nmol/L 17β-estradiol treatment had no influence on nNOS/PSD95 conjugating.Conclusion：17β-estradiol of 10 μmol/L could injure cortical hippocampal neurons by promoting the conjugating of nNOS and PSD95.