骨形态发生蛋白4对胎鼠脑神经元胆碱能表达上调的效应  

Up-regulation effect of bone morphogenetic protein 4 on cholinergic-expression for neurons of embryo rats

作　　者：罗芸[1] 刘磊[1] 薛毅珑[1] 李雁凌[1] 崔忻[1] 潘静坤[1] 

机构地区：[1]解放军总医院老年医学研究所,北京100853

出　　处：《解放军医学杂志》2005年第9期825-827,共3页Medical Journal of Chinese People's Liberation Army

摘　　要：目的观察了骨形态发生蛋白4(BMP4)对原代分离培养的大鼠脑海马和大脑皮质的神经元胆碱能表达的调节效应。方法将从胎龄2周内大鼠脑海马和大脑皮质分离的细胞培养于含AraC的DMEM/F12培养液中,在光镜下观察细胞的形态特征,并行微管相关蛋白(MAP2)细胞化学染色鉴定。AraC作用96h后,改换成含有BMP4的培养液,继续培养10天,行胆碱乙酰转移酶(ChAT)免疫细胞化学染色和流式细胞术检测。结果从胎龄大鼠海马和大脑皮质分离出的细胞具有典型神经元形态学特征和免疫学标志;加入BMP4组的ChAT阳性细胞比率显著高于未加BMP4的对照组(13.3%±1.37%,6.44%±0.81%,P<0.01)。流式细胞术也测得BMP4组的ChAT阳性细胞比率高于未加BMP4的对照组(11.48%、4.92%)。结论BMP4可体外上调胎鼠海马及大脑皮质神经元的胆碱能表达。Objective To Observe the adjusting effect of bone morpbogenetic protein 4 （BMP4） on cholinergic-expression in embryo rats＇ neurons. Method Neurons isolated from hippocampus and cerebral cortex of 15-20 days gestational age SD rat＇s brain were cultivated in a DMEM/F12 medium containing Ara C and identified with morphological characters and microtubulin associated protein 2 （MAP2） immunocytochemical test. The medium was replaced with BMP4 96 hours after Ara C adding, and cultivated another 10 days. The expression of choline acetyhransferase （CHAT） of the adjusted cells was observed by irnmunocytochemical staining and FITC-labeled cholinergic nerve cells Flow Cytometry. Results It has been showed that cultured neurons accorded with the typical morphological characters and the immunologic markers of neurons; the percentage of ChAT-positive cells in BMP4 group was prominently higher than that in control group（13. 3 ±1.37% vs 6. 44±0. 81%, P〈0. 01）, and the same result was showed by Flow Cytometry （11.48% vs 4. 92%）. Condusion BMP4 may in vitro up-regulate the cholinergic expression of the neurons in hippocampus and cerebral cortex of embryo rats.

关 键 词：骨形态发生蛋白 神经元 上调 胆碱能 

分 类 号：R338.14[医药卫生—人体生理学]