ERK1／2信号通路对丹参诱导骨髓基质细胞向神经元样细胞分化的影响

Influence of ERK 1/2 signal-transduction pathway in differentiation of bone marrow stroma cells induced by Salvia miltiorrhiza

目的研究细胞外信号调节蛋白激酶1／2（ERK1／2）信号通路对丹参制剂诱导大鼠骨髓基质细胞（BMSCs）向神经元样细胞分化过程的影响。方法分离、纯化大鼠BMSCs，取第3代细胞进行诱导分化。根据诱导液不同分为空白对照组（A组）、丹参制剂组（B组）及PD98059（ERK1／2特异性抑制剂1组（C组）。对各组细胞进行形态学观察，免疫组织化学方法检测神经元特异性烯醇化酶（NSE）、巢蛋白的蛋白表达，PCR检测NSE和巢蛋白的mRNA的表达。结果诱导1h后，A组细胞形态变化不明显；B组可见细胞形态发生改变，少数细胞呈锥形，细胞突起增多伸长，继续诱导培养5h后细胞突起增多，呈类神经元细胞分化，突起间相互交织；C组细胞总数、类神经元细胞和突起数目均较B组减少。诱导5h后，B组NSE和巢蛋白染色细胞阳性率和mRNA表达量较A、C组明显增高。差异均有统计学意义（P〈0．05）。结论丹参制剂可以促进BMSCs向神经元样细胞分化。ERK1／2的特异性抑制剂PD98059能够拮抗丹参制剂促分化作用，提示ERKl／2信号通路可能参与了丹参制剂诱导BMSCs向神经元样细胞分化的过程。

Objective To study the effect of extracellular signal-regulated ldnase 1/2 （ERK1/2） signal transduction pathway on differentiation of rat bone marrow stroma cells （BMSCs） into neuron-like cells induced by Salvia miltiorrhiza. Methods Rat BMSCs were isolated and purified in vitro by whole bone marrow method. At the third passage, BMSCs were induced and assigned into 3 groups： control group （group A）, Salvia miltiorrhiza treatment group （group B） and PD98059 （ERK1/2 specific inhibitor） treatment group （group C）. Morphology was observed. Neuron-specific enolase （NSE） and Nestin protein expressions were detected by immunohistochemistry. The mRNA expressions of NSE and Nestin were detected by PCR. Results Following 1 h of induction, no significant changes in cell morphology was noted in group A; cells in group B showed obvious changes, with some cone-shaped and with an increased number of cells which stretched and had similar shape with neurons, and cells stretched much more obviously over time and connected to each other; the number of total cells, neuron-like cells and neuritis in group C was decreased as compared with those in group B. As compared with group A and group C, group B had significantly higher NSE- and Nestin-positive rates, and higher NSE and Nestin mRNA expressions 5 h after treatment （P〈0.05）. Conclusions Salvia miltiorrhiza can promote the differentiation of BMSCs into neuron-like ceils. ERK1/2 specific inhibitor PD98059 can resist Salvia miltiorrhiza effects, indicating that ERK1/2 is the key signal pathway of the promotion effects of Salviamiltiorrhiza on differentiation of BMSCs into neuron-like cells.