CAJ | 학술논문

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)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,ERK 1/2的特异性抑制剂PD98059能够拮抗丹参制剂促分化作用,提示ERK1/2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.
목적 연구세포외신호조절단백격매1/2(ERK1/2)신호통로대단삼제제유도대서골수기질세포(BMSCs)향신경원양세포분화과정적영향. 방법 분리、순화대서BMSCs,취제3대세포진행유도분화.근거유도액불동분위공백대조조(A조)、단삼제제조(B조)급PD98059(ERK1/2특이성억제제)조(C조).대각조세포진행형태학관찰,면역조직화학방법검측신경원특이성희순화매(NSE)、소단백적단백표체,PCR검측NSE화소단백적mRNA적표체. 결과 유도1h후,A조세포형태변화불명현;B조가견세포형태발생개변,소수세포정추형,세포돌기증다신장,계속유도배양5h후세포돌기증다,정류신경원세포분화,돌기간상호교직;C조세포총수、류신경원세포화돌기수목균교B조감소.유도5h후,B조NSE화소단백염색세포양성솔화mRNA표체량교A、C조명현증고,차이균유통계학의의(P＜0.05). 결론 단삼제제가이촉진BMSCs향신경원양세포분화,ERK 1/2적특이성억제제PD98059능구길항단삼제제촉분화작용,제시ERK1/2신호통로가능삼여료단삼제제유도BMSCs향신경원양세포분화적과정.
Objective To study the effect of extracellular signal-regulated kinase 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 cells.ERK1/2 specific inhibitor PD98059 can resist Salvia miltiorrhiza effects,indicating that ERK1/2 is the key signal pathway of the promotion effects of Salvia miltiorrhiza on differentiation of BMSCs into neuron-like cells.