摘要
目的：观察滋补肝肾中药六味地黄丸体外诱导大鼠BMSCs分化为神经元样细胞的作用,为进一步探索获得较高比例神经元样细胞的诱导方法打下基础。方法：采用全骨髓贴壁分离法分离、培养SD大鼠BMSCs,鉴定后的第3代细胞经bFGF预诱导24 h后按空白组、bFGF组、六味地黄丸组进行诱导,观察细胞形态变化,诱导培养7 d,采用免疫细胞化学染色法检测相关标志物巢蛋白（Nestin）、神经元特异性烯醇化酶（NSE）、胶质纤维酸性蛋白（GFAP）的表达。结果与结论：bFGF组、六味地黄丸组部分细胞逐渐伸出突起,多为2~3极,呈现较典型的神经元样细胞形态;均有Nestin、NSE、GFAP阳性表达,六味地黄丸组Nestin、NSE表达阳性率高于bFGF组,差异有统计学意义（P〈0.05）。结果说明：（1）全骨髓贴壁分离法分离、培养的骨髓间充质干细胞能逐渐纯化,细胞活性较高,生物学性状稳定,适于做进一步研究;（2）六味地黄丸能诱导骨髓间充质干细胞向神经元样细胞定向分化,可继续探讨六味地黄丸和碱性成纤维细胞生长因子联合诱导是否可获得更高比例的神经元样细胞。
Objective： This study was to explore the effect of Chinese medicine Liuwei Dihuang Pill which can nourish liver on the differentiation of rat bone marrow mesenchymal stem cells （ BMSCs ） into neuron-like cells by cell biology techniques. We tried to find a method to revecie a higher proportion of neuron-like cells. Methods ： We cultured the flushing fluid of Sprague-Dawley rat bone marrow and divided the BMSCs into control group, bFGF group, Liuwei Dihuang Pill group. They were induced by adding the corresponding induction medium after pre-induction for 24 hours by bFGF. We observed the morphological changes of the cells in every group, identifying the induced cells by immunocytochemical staining of nestin, neuron-specific enolase （ NSE ）, glial fibrillary acidic protein （ GFAP ） to judge whether the differentiation was occurrence in 7 days later. Results ： After induced by bFGF or Liuwei Dihuang Pill for 7 days, BMSCs formed the more typical form of neural cells. The immunoeytochemical staining results showed that the neural cell makers were not expressed in the control group, but expressed in other groups. Positive cells rate of Nestin and NSE in the Liuwei Dihuang Pill group were higher than those of the bFGF group. Conclusion ： The BMSCs had high activity and stability biological traits which were separated, cultured and purified by the method of bone marrow adherent seperation. The BMSCs could be differentiated into neuron-like cells in vitro with the induction of Liuwei Dihuang Pill. The combined induction of Liuwei Dihuang Pill and bFGF revecied a higher proportion of neuron-like cells.
出处
《辽宁中医药大学学报》
CAS
2014年第10期37-40,共4页
Journal of Liaoning University of Traditional Chinese Medicine
作者简介
龚圆渊（1986-），女，四川绵阳人，助教，硕士，研究方向：神经解剖与临床。
