皮质酮对体外培养发育中胎鼠脑皮层神经元迁移蛋白LIS1表达的影响

Effect of corticosterone on lissencephaly 1 expression in developing cerebral cortical neurons of fetal rats cultured in vitro

摘要:

目的 研究皮质酮对发育中胎鼠脑皮层神经元迁移蛋白Lissencephaly 1（LIS1）表达的影响。方法 体外原代培养Wistar胎鼠脑皮层神经元并分为对照组、低浓度干预组和高浓度干预组，各组给予含有不同浓度皮质酮（对照组：0 μmol/L，低浓度干预组：0.1 μmol/L，高浓度干预组：1.0 μmol/L）的培养基干预，分别于干预后1、4、7 d收集细胞，采用Werstern blot法及免疫细胞化学染色法观察神经元LIS1蛋白表达的变化。结果 Western blot结果显示干预后7 d，低浓度干预组及高浓度干预组胎鼠脑皮层神经元胞浆及胞核内LIS1表达均明显低于对照组（P < 0.05），且高浓度干预组胞浆内LIS1表达明显低于低浓度干预组（P < 0.05）。免疫细胞化学染色法结果显示皮质酮干预后1、4、7 d，高浓度干预组LIS1平均光密度值明显低于对照组及低浓度干预组（P < 0.05），且干预后7 d，低浓度干预组LIS1平均光密度值明显低于对照组（P < 0.05）。结论 皮质酮可下调体外培养发育中胎鼠脑皮层神经元迁移蛋白LIS1的表达水平，且与皮质酮浓度高低、作用时间长短相关。

Abstract:

Objective To investigate the effect of corticosterone on the expression of the neuronal migration protein lissencephaly 1 (LIS1) in developing cerebral cortical neurons of fetal rats. Methods The primary cultured cerebral cortical neurons of fetal Wistar rats were divided into control group, low-dose group, and high-dose group. The neurons were exposed to the medium containing different concentrations of corticosterone (0 μmol/L for the control group, 0.1 μmol/L for the low-dose group, and 1.0 μmol/L for the high-dose group). The neurons were collected at 1, 4, and 7 days after intervention. Western blot and immunocytochemical staining were used to observe the change in LIS1 expression in neurons. Results Western blot showed that at 7 days after intervention, the low-and high-dose groups had significantly higher expression of LIS1 in the cytoplasm and nucleus of cerebral cortical neurons than the control group (P < 0.05), and the high-dose group had significantly lower expression of LIS1 in the cytoplasm of cerebral cortical neurons than the low-dose group (P < 0.05). Immunocytochemical staining showed that at 1, 4, and 7 days after corticosterone intervention, the high-dose group had a significantly lower mean optical density of LIS1 than the control group and the low-dose group (P < 0.05). At 7 days after intervention, the low-dose group had a significantly lower mean optical density of LIS1 than the control group (P < 0.05). Conclusions Corticosterone downregulates the expression of the neuronal migration protein LIS1 in developing cerebral cortical neurons of fetal rats cultured in vitro, and such effect depends on the concentration of corticosterone and duration of corticosterone intervention.