氰戊菊酯通过干扰雌激素作用引起小鼠海马神经细胞损伤

Fenvalerate induce hippocampal neurons injury through interfering with estrogen action

目的 探讨氰戊菊酯（fenvalerate,FEN）能否通过干扰雌激素（E2）的作用引起小鼠海马神经细胞损伤.方法 取孕18d的ICR小鼠胚胎海马细胞原代培养,建立小鼠海马神经元和星形胶质细胞的混合培养模型.以氰戊菊酯（FEN,0、1、10、50 μg/ml）,FEN（0、10、50 μg/ml）联合雌激素受体抑制剂（ICI 182,780,1μmol/L）,FEN（0、10、50 μg/ml）联合E2（10 nmol/L）对体外培养7d后的细胞染毒48 h.采用免疫荧光细胞化学方法检测神经元和星形胶质细胞的相对数量,测量神经元的突起长度.结果 1 μg/ml FEN组混合培养的小鼠海马神经元数量、神经突起和原浆型星形胶质细胞均无明显改变,10和50μg/ml FEN组海马神经元数目减少,突起长度缩短,原浆型星形胶质细胞比例增加,与对照组比较,差异有统计学意义（P＜0.05）.ICI 182,780单独作用组神经元数量、突起长度和原浆型星形胶质细胞均无明显改变.ICI＋ 10 μg/ml FEN组海马神经元数目增加,突起长度延长,原浆型星形胶质细胞比例减少,与10 μg/ml FEN单独作用组比较,差异有统计学意义（P＜0.05）; ICI＋50 μg/ml FEN组海马神经元数目增加,原浆型星形胶质细胞比例减少,与50 μg/ml FEN单独作用组比较,差异有统计学意义（P＜0.05）.E2单独组神经元数目增加,神经突起长度延长,与对照组比较,差异有统计学意义（P＜0.05）;E2＋10 μg/ml FEN组和E2＋ 50μg/ml FEN组神经元数目减少,突起长度缩短,原浆型星形胶质细胞比例增加,与E2单独作用组比较,差异有统计学意义（P＜0.05）.结论 FEN可通过干扰E2的经典核受体信号通路引起混合培养的海马神经元丢失,可通过干扰E2的经典核受体信号和膜受体通路抑制神经突起生长,FEN对E2的干扰效应在其神经发育毒性中起重要作用.

Objective To investigate whether fenvalerate can induce mouse hippocampal nerve cell damage by interfering with estrogen （E2） effect.Methods Hippocampus were dissected and cultured from Embryo 18 d ICR mice,the cells were cultured for 7 days.Fenvalerate （FEN,0,1,10,50 μg/ml）,FEN（10,50 μg/ml） and estrogen receptor antagonist ICI 182,780 （1 μmol/L）,FEN （0,10,50 μg/ml） and E2 （10 nmol/L） were applied to the cultured cells for 48h.Immunocytochemically stained with neurons and astrocytes to evaluate the levels respectively,and the growth of neurite.Result 1μg/ml FEN have no effect on neurons,neurites and protoplasmic astrocytes,10 and 50 μg/ml FEN can significantly decrease the neuron viability and the length of neurite as well as increase the level of protoplasmic astrocytes （P〈0.05 vs.control group）.ICI 182,780 alone have no effect on neurons,neurites and protoplasmic astrocytes; ICI＋10 μg/ml FEN significantly increase the cell viability and extend neurite length as well as decrease protoplasmic astrocytes （P〈0.05 vs.10 μg/ml FEN alone group）; ICI＋50 μg/ml FEN significantly increase the cell viability and decrease protoplasmic astrocytes （P〈0.05 vs.50 μg/ml FEN alone group）.E2 alone have no effect on protoplasmic astrocytes,while can promote neuronal survival and neurite growth; E2＋10 μg/ml FEN and E2＋50 μg/ml FEN significantly decrease neuronal survival and neurite growth,as well as increase protoplasmic astrocytes （P〈0.05 vs.E2 alone group）.Conclusion Fenvalerate can induce the loss of hippocampal neurons through disrupting estrogen nuclear receptor signaling,and inhibit the length of neurite through disrupting estrogen nuclear receptor and membrane receptor signaling.The effect of estrogen disruption play an important role in developmental neurotoxicity by fenvalerate.