大鼠皮质神经细胞缺氧缺糖损伤后MAP-2表达变化规律

Oxygen-Glucose deprivation-induced dynamic changes of MAP-2 in cultured rat cortical neurons

目的观察原代培养大鼠大脑皮质神经细胞缺氧缺糖（OGD）损伤24h内的形态学及微管相关蛋白2（MAP-2）表达变化。方法使用出生1-2d的Wistar大鼠,取大脑皮质神经细胞进行原代培养,用连二亚硫酸钠加低糖培养基制作神经细胞OGD损伤模型,损伤后应用倒置显微镜观察形态学变化,Western印迹法观察MAP-2蛋白表达变化。结果原代培养大鼠大脑皮质神经细胞OGD损伤后1h,少数神经细胞突起缩短;伤后6-12h神经细胞胞体明显皱缩,多数细胞突起缩短或消失;伤后24h神经细胞形态部分恢复,部分突起重新出现。Western印迹法显示,MAP-2在神经细胞OGD损伤后1h表达降低,伤后12h达低谷,24h表达升高。结论 OGD损伤后,原代培养大鼠大脑皮质神经细胞的形态及MAP-2表达随时间呈现一定的规律性变化。

Objective To study the expression of microtubule-associated protein-2（ MAP-2） and morphological changes of cultured rat cortical neurons after oxygen-glucose deprivation（ OGD） injury.Methods Cortical neurons were isolated from P1 - 2 neonatal Wistar rat cerebral cortices and cultured in OGD medium. MAP-2 expression and morphological changes of the cultured neurons were investigated by Western Blotting and under inverted microscope. Results Without OGD injury,the cultured neuron had round or elliptical cell body with long intact dendrites. The neurons remained round with short dendrites at1 h post-OGD,while wrinkled with few dendrites at 6h and 12 h post-OGD; the neurons partly recovered and some dendrites reappeared at 24 h post-OGD. Western Blotting showed MAP-2 protein decreased 1h post-OGD,reached the minimal level at 12 h,and then increased at 24 h post-OGD. There was no significant difference between MAP-2 expressed at 6h post-OGD and 12 h post-OGD. Conclusion After moderate OGD injury,cultured rat cortical neurons demonstrate time-dependent MAP-2 expression and morphological changes.