EGCG对体外培养胚鼠皮层神经细胞的保护作用

The Effects of Green Tea Polyphenol (-) Epigallocatechin Gallate on the Growth of Mice Cortical Neurons in Vitro

目的观察表没食子儿茶酚没食子酸酯(EGCG)对体外培养的胚鼠皮层神经细胞生长的影响。方法离体培养昆明种小鼠胚鼠大脑皮层神经细胞,至第5天加入不同浓度的EGCG(1、5、10、20、40、60、80和100μmol/L),通过倒置相差显微镜和NSE免疫细胞化学染色法进行神经元形态观察,应用中性红比色法和MTT法测定神经细胞的存活数和细胞活性。结果EGCG实验组培养48h后,与对照组比较,能明显促进神经细胞存活及突起生长,可增加神经细胞的存活数,提高神经细胞内酶的活性,尤其在10～40μmol/L的浓度范围内差异较显著(P<0.01)。结论在5～100μmol/L的浓度范围内,EGCG均有一定的促细胞生长作用,可增加神经细胞的存活数,提高神经细胞内酶的活性。

Objective To investigate the effects of epigallocatechin gallate （EGCG）, an abstract from chinese medical herbs,green tea polyphenol,on the growth of mice cortical neurons in vitro. Methods Neurons were dissociated from the cerebral cortex of embryonic mice and cultured in DMEM with 20% FCS. In the fifth day, the neurons were treated with 1,5, 10,20, 40,60,80 and 100μmol/L EGCG, respectively. The process of the growth and morphology of the neurons were observed under phase contrast microscope in vitro. The cultured cortical neurons were stained with anti- NSE antibody and the cell survival was analyzed using colormetric neutral red assay, as well as the cell viability was determined by MTT assay. Results EGCG could enhance the survival of neurons and improve the viability of normal neurons with concentration dependent way at 1,5,10,20,40, 60,80 and 100μmot/L for 48hr. In contrast with the control groups,the most significant effective range of concentration on neuronal survival was from 10 to 40μmol/L（ P 〈 0.01） ,and 5,60,80 and 100μmol/L were significantly effective （ P 〈 0.05 ）, respectively. Conclusion Supplementation of suitable amount of ECCG （5-10μmol/L） can improve the growth of cortical neurons in vitro, enhance the mouse cortical neuronal survival, as well as improve cortical neurons viability.