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目的:研究人参皂甙Rg1对小鼠脂肪干细胞神经样分化的促进作用,并初步探讨其作用机理。创新点:证明了人参皂甙Rg1可以通过mi RNA-124途径促进脂肪干细胞的神经样分化。方法:从BALB/c小鼠腹股沟和睾丸脂肪垫处分离培养脂肪干细胞,利用流式细胞仪检测分离的脂肪干细胞纯度。试验分为以下五组:磷酸缓冲盐溶液(PBS)组、3-异丁基-1-甲基黄嘌呤(IBMX)组、IBMX+Rg1低剂量组、IBMX+Rg1中剂量组和IBMX+Rg1高剂量组。用细胞免疫组化方法检测了脂肪干细胞向神经样细胞的分化效率,用荧光定量聚合酶链式反应(qP CR)方法检测mi RNA-124的表达变化,用免疫印迹的方法检测巢蛋白(nestin)、βIII-微管蛋白(βIII-tubulin)及羧基端小结构域磷酸酶1(SCP1)的表达水平。结论:免疫组化结果显示,IBMX可以成功诱导小鼠脂肪干细胞向神经样细胞的分化;免疫印迹结果显示,Rg1可以显著提高神经样细胞标记蛋白的表达水平;荧光定量PCR结果显示,Rg1可以促进miRNA-124的表达量,进而降解神经分化抑制因子SCP1的表达,促进脂肪干细胞的神经样分化效率。
OBJECTIVE: To study the effects of ginsenoside Rg1 on the neuronal differentiation of mouse adipose-derived stem cells and to explore its mechanism. Innovative point: Proved that ginsenoside Rg1 can promote neural differentiation of ADSCs through the mi RNA-124 pathway. Methods: Adipose-derived stem cells were isolated and cultured from the groin and testicular fat pads of BALB / c mice. The purity of isolated adipose-derived stem cells was detected by flow cytometry. The experiments were divided into the following five groups: phosphate buffered saline (PBS) group, 3-isobutyl-1-methylxanthine (IBMX) group, IBMX + Rg1 low dose group, IBMX + Rg1 medium dose group and IBMX + Rg1 High-dose group. The differentiation efficiency of adipose-derived stem cells to neuron-like cells was detected by immunohistochemistry. The expression of mi RNA-124 was detected by qPCR method. The expression of nestin ), ΒIII-tubulin and small carboxyl-terminal domain phosphatase 1 (SCP1). Conclusion: The results of immunohistochemistry showed that IBMX can successfully induce the differentiation of ADSCs into neuron-like cells. Immunoblotting showed that Rg1 could significantly increase the expression of neuron-like marker protein. Fluorescent quantitative PCR showed that Rg1 could promote miRNA-124 expression, and then degradation of the expression of neural differentiation inhibitor SCP1, and promote the efficiency of ADSCs.