补肾填精法促进骨髓基质干细胞分化为神经元样细胞的实验研究

目的:探讨补肾填精方右归丸含药血清体外诱导大鼠骨髓基质干细胞分化为神经元样细胞的作用.方法:用DMEM/F-12培养液培养Wistar大鼠骨髓基质干细胞(BMSCs).用右归丸含药血清诱导BMSCs分化后,倒置显微镜观察细胞形态变化；用尼氏染色法观察神经细胞特征性结构；用免疫细胞化学染色法鉴定神经胶质细胞标志物胶质纤维酸性蛋白(GFAP)及神经细胞特异性标志物神经元特异性烯醇化酶(NSE),微管相关蛋白-2(MAP-2)的表达.结果:诱导6h后,骨髓基质干细胞胞体收缩,有突起伸出；诱导24 h后突起增多形成网络结构；尼氏染色法显示诱导后的细胞胞体中有尼氏体.免疫细胞化学染色示NSE及MAP-2阳性表达,GFAP无表达.结论:右归丸含药血清可以体外诱导大鼠骨髓基质干细胞分化为神经元样细胞.

Experiment Research of Inducing Bone Marrow Stem Cells to Differentiate into Neurons-like Cells by Way of Tonifying Kindey Essence

Objective: To discuss the effect of the serum containing Yougui Pill in inducing bone marrow stem cells to differentiate into neurons-like cells in vitro. Method: Rat bone marrow stromal stem cells (BMSCs) was cultured in DMEM/F-12 medium and inducted into neurons-like cells by serum containning Yougui Pill. The morphologic changes were observed by light microscope, the specific characteristic structures of neurons cells were observed using Nissl's staining method,gial fiber acidic protein(GFAP), neuron specific enolase(NSE) and microtubule-associated protein-2(MAP-2) were detected by immunohistochemistry. Result: By induction, BMSCs' body shrinked and showed processes after 6 hours; the prominences interlaced to net, forming typical nerve fiber net after 24 hours; trachychromatic Nissl body was found in cytolymph. The NSE and MAP-2 were determined by immunohistochemistry, GFAP was undetermined. Conclusion: BMSCs could be inducted into neurons-like cells in vitro by serum containing Yougui Pill.