地黄饮子含药血清对大鼠BMSCs与NSCs共培养影响转归的研究  

Effect of Rehmannia Decoction Contained Serum on NSCs of Rats in BMSCs and NSCs Co-culture System

作　　者：武博文[1] 武密山[1,2] 王慧娜 郭金栋 高维娟 王茹[1] 韩红伟 师旭亮[1] 

机构地区：[1]河北中医学院,河北石家庄050200 [2]河北省心脑血管病中医药防治重点实验室,河北石家庄050091 [3]承德医学院,河北承德067000

出　　处：《湖南中医药大学学报》2017年第10期1056-1062,共7页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金资助项目(81073074;30472200);河北省自然科学基金项目(H 2013206005);河北省高等学校科学技术研究重点项目(ZD2015053);河北省高等学校高层次人才科学研究项目(GCC2014013)

摘　　要：目的探讨以升麻为药引子的地黄饮子(CRD)含药血清对经过骨髓间充质干细胞(BMSCs)共培养后的大鼠神经干细胞(NSCs)的存活和分化的影响。方法建立BMSCs与NSCs共培养模型,在共培养体系中加入CRD含药血清,从培养第3天开始分别进行5-溴-2-脱氧尿嘧啶(Brd U)和神经元特异性烯醇化酶(NSE)、胶质细胞纤维酸性蛋白(GFAP)、神经元特异性标志物微管相关蛋白2(MAP-2)、少突胶质细胞特异性表达蛋白标志物(MBP)免疫细胞化学双染色。采用流式细胞仪检测NSCs的存活率,利用细胞免疫荧光和Western blot法鉴定NSCs的分化情况,比较NSCs的存活率和分化率,分析CRD含药血清对共培养后NSCs的存活及分化的影响。结果与生理盐水对照组相比,中、高浓度CRD含药血清在BMSCs与NSCs共培养体系中第7天能显著提高NSCs存活率和NSE、GFAP、MAP-2、MBP活性(P<0.01)。结论 CRD含药血清在BMSCs与NSCs共培养体系中能促进NSCs有效分化和提高NSCs存活率。Objective To explore the effect of Cimicifuga heracleifolia Kom. as medicinal guiding of Rehmannia decoction（CRD） contained serum on survival and differentiation of neural stem cells（NSCs） in NSCs and bone marrow mesenchymal stem cells（BMSCs） co-culture system. Methods Firstly, the modeling of BMSCs co-cultured with NSCs was set up,then CRD contained serum was put into this co-culture syste. The cells of 5-bromo-2-deoxyuridine（Brd U） and neuron-specific enolase（NSE）, glial fibrillary acidic protein（GFAP）, microtubule-associated protein（MAP2）, myelin basic protein（MBP） were stained with double immunocytochemical staining method at 3 rd dayof culture. The survival rate of NSCs was detected by the flow cytometry（FCM） and expression of NSCs was detected by immunocytochemistry and Western bolt. The effect of CRD contained serum on survival and differentiation of NSCs was analyzed. Results Compared with the normal saline group, higher concentration and middle concentration of CRD contained serum could significantly increased the survival rate of NSCs, and the NSE, GFAP, MAP-2 and MBP activity on the 7 th day in BMSCs and NSCs co-culture system（P〈0.01）. Conclusion CRDcontained serum could promote the effective differentiation and increase the survival rate activity of NSCs and in BMSCs and NSCs co-culture system.

关 键 词：骨髓间充质干细胞 神经干细胞 药引子 地黄饮子 含药血清 神经样细胞 

分 类 号：R285.5[医药卫生—中药学] R393[医药卫生—中医学]