Caveolin-1在大鼠骨髓间质干细胞分化为神经细胞中的作用

发布时间：2018-06-03 20:46

本文选题：骨髓间质干细胞 + 神经元　；参考：《中国病理生理杂志》2010年07期

【摘要】：目的:探讨caveolin-1在骨髓间质干细胞(MSCs)分化为神经细胞中的作用。方法:实验分为未转染组、转染组(转染Rn-caveolin-1-siRNA)、阳性对照组(转染Rn-MAPK-1control siRNA)及阴性对照组(转染negative control siRNA)4组。采用β-巯基乙醇诱导大鼠MSCs分化为神经细胞。倒置荧光显微镜下观察MSCs转染后荧光表达情况;RT-PCR检测caveolin-1和促分裂原活化蛋白激酶1(MAPK1)mRNA的表达变化;免疫细胞化学法检测caveolin-1、神经元烯醇化酶(NSE)、神经微丝亚单位(NF-M)和胶质纤维酸性蛋白(GFAP)的表达变化;MTT方法检测细胞存活率。结果:(1)siRNA转染72h,MSCs荧光表达最强,转染率可达(81.5±2.8)%;而且转染组MSCs的caveolin-1mRNA转录下降(P0.05);MTT提示转染组细胞存活率无显著变化(P0.05)。(2)β-巯基乙醇可以诱导MSCs向神经细胞分化,其中以转染组诱导效果最佳,NSE、NF-M的表达率显著高于其它各组(P0.01)。(3)随诱导时间延长,各组caveolin-1表达持续增加,诱导6d达到峰值,与诱导前、诱导6h相比有显著差异(P0.05)。此外,转染组与其它各组同时点的caveolin-1表达均显著降低(P0.01)。结论:以caveolin-1为标记蛋白的脂筏在MSCs诱导分化为神经细胞中可能起到重要的调控作用。
[Abstract]:Objective: to investigate the role of caveolin-1 in differentiation of bone marrow mesenchymal stem cells into neural cells. Methods: the experiment was divided into non-transfection group, transfected Rn-caveolin-1-siRNA-transfected group, positive control group (Rn-MAPK-1control siRNA-transfected group) and negative control group (transfected negative control siRNA)4 group). Rat MSCs was induced to differentiate into nerve cells by 尾-mercaptoethanol. Reverse fluorescent microscope was used to observe the expression of MSCs after transfection. RT-PCR was used to detect the expression of caveolin-1 and mitogen-activated protein kinase (1(MAPK1)mRNA). The expression of caveolin-1, neuronal enolase NSE, neurofilament subunit (NF-M) and glial fibrillary acidic protein (GFAP) were detected by immunocytochemistry. Results the fluorescence expression of MSCs was the strongest in 72hs transfected with MSCs, the transfection rate was 81.5 卤2.8, and the caveolin-1mRNA transcription of MSCs in transfection group was decreased (P0.05). The results showed that there was no significant change in cell survival rate in transfected group. 尾 -mercaptoethanol could induce the differentiation of MSCs into neural cells. The expression rate of NSE-NF-M in the transfection group was significantly higher than that in the other groups (P 0.01). The expression of caveolin-1 in each group continued to increase and reached its peak value on the 6th day after induction. There was a significant difference between the two groups at 6 h after induction (P 0.05). In addition, the expression of caveolin-1 was significantly decreased in the transfection group and other groups at the same time. Conclusion: lipid rafts with caveolin-1 as a marker may play an important role in the differentiation of nerve cells induced by MSCs.
【作者单位】：郑州大学第一附属医院神经内科;
【基金】：国家自然科学基金资助项目(No.30770758) 河南省教育厅自然科学研究计划资助项目(No.2008A320032)
【分类号】：R329.21

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