大鼠骨髓间充质干细胞向神经细胞分化后自噬水平的变化

发布时间：2018-11-15 10:09

【摘要】：目的:探讨骨髓间充质干细胞(BMSCs)向神经细胞分化前后自噬水平的变化,进一步研究BMSCs神经分化机制。方法:原代培养大鼠BMSCs,三代后用L-DMEM(2%DMSO+200μmol/L BHA)诱导其向神经细胞分化,采用免疫细胞化学方法检测神经元特异标记物NSE表达,采用RT-PCR方法检测自噬相关基因LC3B、Beclin1、Atg5、Atg7、Atg10的表达,用Western blot及流式细胞术检测了自噬标志蛋白LC3B的表达。结果:大鼠BMSCs传3代后,贴壁的BMSCs可达90%以上,类成纤维细胞样梭形生长,经诱导剂作用5 h后,细胞呈现神经细胞样改变,NSE阳性率为(83±5)%。RT-PCR结果表明BMSCs向神经细胞分化后LC3B,Beclin1,Atg5,Atg7,Atg10的表达升高,流式细胞术检测LC3B的平均荧光强度升高,Western blot结果表明神经分化后LC3B-Ⅱ表达增加。结论:大鼠BMSCs向神经细胞分化后自噬水平增加。
[Abstract]:Aim: to investigate the changes of autophagy level before and after (BMSCs) differentiation in bone marrow mesenchymal stem cells (BMSCs) and to further study the mechanism of BMSCs neural differentiation. Methods: rat BMSCs, was induced to differentiate into neural cells by L-DMEM (2%DMSO 200 渭 mol/L BHA) for three generations. The expression of neuron specific marker NSE was detected by immunocytochemistry. The expression of autophagy associated gene LC3B,Beclin1,Atg5,Atg7,Atg10 was detected by RT-PCR and the expression of autophagy marker protein LC3B was detected by Western blot and flow cytometry. Results: after 3 passages of rat BMSCs, more than 90% of BMSCs adherent to the wall. Fibroblast-like fusiform growth was observed. After 5 h of induction, the cells showed neuronal changes. The positive rate of NSE was (83 卤5)%. The results of RT-PCR showed that the expression of LC3B,Beclin1,Atg5,Atg7,Atg10 increased after the differentiation of BMSCs into neurons, and the mean fluorescence intensity of LC3B was increased by flow cytometry. Western blot results showed that the expression of LC3B- 鈪，

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