Cyclin D1在黄芪甲苷诱导BMSCs向神经元样细胞分化中的表达  

Cyclin D1 Expression During Astragaloside A Induced Neuron-Like Cell Differentiation of BMSCs

作　　者：王晓寅[1] 王会芳[1] 罗强 张辉 张晓丽 WANG Xiao-yin;WANG Hui-fang;LUO Qiang;ZHANG Hui;ZHANG Xiao-li(The Second Affiliated Hospital of HeBei North University,Zhangjiakou,075000,China;Life Science Research Center of HeBei North University,Zhangjiakou,075000,China)

机构地区：[1]河北北方学院附属第二医院,张家口075000 [2]河北北方学院生命科学研究中心,张家口075000

出　　处：《神经药理学报》2024年第1期11-17,共7页Acta Neuropharmacologica

基　　金：河北省教育厅重点项目(No.ZD2014067)。

摘　　要：目的:研究黄芪甲苷诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分化的特点和诱导过程中Cyclin D1的表达特点。方法:采用100 mg·L^(-1)黄芪甲苷诱导BMSCs分化,在1、12、24 h倒置显微镜观察细胞形态变化,免疫细胞化学方法观察巢蛋白(Nestin)、神经元特异性烯醇化酶(neuron specific enolase,NSE)、Cyclin D1的表达,RT-PCR检测Cyclin D1 mRNA的表达。结果:收集获得的骨髓细胞采用原代和传代培养,至第3代BMSCs细胞基本纯化。经100 mg·L^(-1)黄芪甲苷作用后,部分细胞自胞体长出突起,形态似神经元样细胞且表达Nestin和NSE抗体,Nestin的表达时相早于NSE且表达强度高于NSE(P<0.05)。在药物作用1 h后,BMSCs细胞Cyclin D1不论在基因转录水平还是在蛋白表达水平均出现大幅度下调,此时BMSCs开始有Nestin的表达;药物作用12、24 h,BMSCs细胞Cyclin D1表达与药物作用前表达水平无显著性差异(P>0.05)。结论:黄芪甲苷可能首先通过调整细胞周期,延长了G1期向S期转变的时间,从而启动了BMSCs的分化程序,诱导BMSCs向神经干细胞分化。在黄芪甲苷的营养作用下,细胞的突起进一步生长并向神经元分化。Objective:To observe the differentiation of bone marrow mesenchymal stem cells(BMSCs)induced by astragaloside A and expression of Cyclin D1.Methods:BMSCs were induced by astragaloside A with 100 mg·L-1.At l,12,24 h after induetion,the morphological changes of BMSCs were observed with optical microscope and the expressions of Nestin,NSE,Cyclin D1 were identified by immunocytochemical techniques.Cyclin D1 mRNA was detected by RT-PCR.Results:Bone marrow cells were primarily cultured and subcultured.The third passage of BMSCs was purified.After treatment with 100 mg·L^(-1) astragaloside A,some cells presented processes,were similar to neuron-like cells in morphology,and expressed Nestin and neuron-specific enolase.Nestin expression occurred earlier(P<0.05)and at a higher level(P<0.05)than expression of neuron-specific enolase.At 1 hour after treatment with astragaloside A,cyclin D1 expression was substantially down-regulated in BMSCs at both gene and protein levels.Simultaneously,BMSCs began to express nestin.No significant differences in cyclin D1 expression in BMSCs were detected between 12 and 24 hours post-treatment and the pre-treatment level(P>0.05).Conclusion:Our results indicated that astragaloside A regulated cell cycle,prolonged G1/S phase transition,triggered BMSC differentiation,and finally induced BMSC differentiation into neuronal stem cells.The trophism of astragaloside A accelerated further growth of cell processes and the differentiation of cells into neurons.

关 键 词：黄芪甲苷 BMSCS 分化 神经元样细胞 Cyclin D1 

分 类 号：R964[医药卫生—药理学]