不同浓度孕酮对Aβ致伤神经干细胞向胆碱能神经元分化的影响  

Effects of progesterone with different concentrations on the differentiation of neural stem cells from Aβ induced neural stem cells to cholinergic neurons

作　　者：徐甜颖 冯莹[1] 程莲芝[1] 肖韩艳[1] XU Tian-ying;FENG Ying;CHEN Lian-zhi;XIAO Han-yan(Department of Neurology, The Second affiliated Hospital, Mudanjiang Medical College, Mudanjiang Heilongiia 15700, China)

机构地区：[1]牡丹江医学院第二附属医院神经内科,黑龙江牡丹江15700

出　　处：《中国卫生工程学》2018年第2期195-198,共4页Chinese Journal of Public Health Engineering

基　　金：牡丹江医学院2016科研项目(zs201521)

摘　　要：目的研究神经甾体孕酮对Aβ_(25-35)损伤后神经干细胞分化的影响。方法制备神经干细胞悬液,设立只加入培养液的空白对照组和只加入Aβ_(25-35)5μmol/L的AD模型组,在Aβ_(25-35)损伤细胞模型中加入0.01、0.1、1、10、100 pmol/L的孕酮共同培养设为梯度孕酮组,应用倒置显微镜及ChAT免疫细胞化学染色观察各组神经干细胞向胆碱能神经元分化的情况。结果不同浓度孕酮组随着浓度升高细胞数目逐渐增多,胞体逐渐饱满,突起逐渐增多而变长;各时间点AD模型组与空白对照组比较ChAT免疫阳性细胞率均明显减少,差异均有统计学意义(均P<0.05);与AD模型组相比,不同浓度孕酮组在各时间点ChAT免疫阳性细胞率均升高,差异均有统计学意义(均P<0.05);在14 d时10 pmol/L浓度孕酮组ChAT免疫阳性细胞率达到(9.04±1.79)%,将14 d时AD模型组及10pmol/L浓度孕酮组进行ChAT、NSE双标免疫细胞荧光染色,结果显示ChAT、NSE双标免疫细胞荧光染色为阳性。结论孕酮可以明显保护Aβ致伤的神经干细胞向胆碱能神经元分化,使胆碱能神经元增多且呈一定浓度依赖性。Objective To study the effect of neurosteroid progesterone on Aβ_（25-35）-induced neural stem cell differentiation.Methods The neural stem cell suspension was prepared,and the blank control group and the AD model group with Aβ_（25-35） 5μmol/L were added only to the culture medium. The model of Aβ_（25-35） injury cell was co-cultured with Aβ_（25-35） cell model with a progesterone of 0. 01,0. 1,1,10,100 pmol/L as a gradient progesterone group. The differentiation of neural stem cells into cholinergic neurons was observed by inverted microscope and chat immunocytochemical staining. Results With the increase of the concentration of progesterone,the number of cells increased gradually,the cell bodies became full,the processes increased and the number of chat immunoreactive cells decreased significantly in the AD model group compared with the blank control group at each time point. The difference was statistically significant（all P〈0.05）. Compared with the AD model group,the rate of chat immunoreactive cells in different concentrations of progesterone group was higher than that in the AD model group at all time points. There were significant differences（P〈0.05）,the rate of chat immunoreactive cells reached（9.04 ±1.79） % in 10 pmol/L progesterone group at 14 days,and ChAT,NSE double labeled immunofluorescence staining was performed in ad model group and 10 pmol/L progesterone group at 14 days. The results showed that the fluorescent staining of ChAT,NSE double labeled immunocytes was positive.Conclusion Progesterone could obviously protect the neural stem cells from Aβ-induced differentiation into cholinergic neurons and increase the number of cholinergic neurons in a concentration-dependent manner.

关 键 词：神经甾体 Β-淀粉样蛋白 神经干细胞 雌激素受体 阿尔茨海默病 

分 类 号：R749.16[医药卫生—神经病学与精神病学]