木犀草素诱导人脐带间充质细胞化为神经细胞潜能的研究  

Research on the potential inducing effect of luteolin on differentiation of human umbilical cord-mesenchymal stem cells into nerve cells in vitro

作　　者：周天秀[1] 陆华[2] 郭丽[1] 任志红[1] 

机构地区：[1]成都中医药大学,成都610037 [2]成都中医药大学第二附属医院,成都610041

出　　处：《中华中医药杂志》2016年第12期5175-5177,共3页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点基础研究发展计划(973计划)项目(No.2010CB530403)~~

摘　　要：目的:探讨木犀草素诱导脐带间充质细胞(h UCMSCs)分化为神经细胞的潜能。方法:采用终浓度为10μg/m L中药单体木犀草素及0.5‰DMSO对人h UCMSCs进行诱导。显微镜下观察细胞形态变化,细胞免疫荧光技术检测神经细胞细胞标记物nestin和GFAP的表达,实时荧光定量PCR测定神经分化相关NSE、nestin、GFAP、MAP2、NEFH 5种基因的表达量变化情况,分析两组数据。结果:木犀草素诱导人h UCMSCs第6天,显微镜下发现胞核增大,细胞呈分叉状改变,细胞伸出较多细长的突起,呈现出神经细胞形态样改变;细胞免疫荧光技术检测显示,实验组大部分细胞神经标记物nestin、GFAP表达阳性,平均荧光分别强度为2.64、4.72,对照组仅有少量细胞nestin、GFAP表达阳性,平均荧光强度分别为0.24、0.46;实时荧光定量PCR结果显示,木犀草素对nestin、GFAP、MAP2、NEFH m RNA的表达有明显影响(P<0.05),对NSE m RNA的表达影响不明显。结论:木犀草素具有诱导h UCMSCs向神经细胞方向分化的潜能。Objective： To investigate the potential inducing effect of luteolin on differentiation of human umbilical cordmesenchymal stem cells（HUCMSCs） into nerve cells. Methods： Ten μg/m L of luteolin and 0.5‰ of DMSO were used to induce h UCMSCs. The change of the cellular morphology was observed by microscopy. The expressions of cellular neuron marker nestin and GFAP were examined by cell immunofluorescence technique. The changes of expression quantity of five genes which related to neural differentiation（NSE, nestin, GFAP, MAP2 and NEFH） were examined by PCR in real time. The data of two groups were analyzed by statistical software SPSS 19.0. Results： The microscopy showed that the forms of the nerve cells have changed after 6 days of intervention. The nucleus was bigger, and the cells had forked changes with many elongated protrusion, which showed morphological changes of never cells. Cellular immunofluorescence technique showed that most cellular neuron markers of nestin and GFAP were positive in the experiment group, and the mean fluorescence intensity were 2.64 and 4.72 respectively. Only few markers were positive in the control group, and the mean fluorescence intensity were only 0.24 and 0.46 respectively. The results of PCR indicated that the intervention by luteolin had significantly influence on the expression of nestin, GFAP, MAP2 and NEFH（P〈0.05）, but the effect on the expression of NSE m RNA was not significant. Conclusion： Luteolin is potential to induce the differentiation of HUCMSCs into nerve cells.

关 键 词：木犀草素 脐带间充质细胞 神经细胞 分化 

分 类 号：R285[医药卫生—中药学]