丹参酮ⅡA诱导骨髓间充质干细胞向心肌细胞分化的研究  

Role of Tanshinone Ⅱ a in inducing differentiation of BMSC into cardiomyocytes

作　　者：宋明官 吕洋[2] 王海萍[1] 任君旭[1] 刘源[1] 

机构地区：[1]河北北方学院基础医学院组织学与胚胎学教研室,张家口075000 [2]河北医科大学基础医学院组织学与胚胎学教研室

出　　处：《中华老年心脑血管病杂志》2015年第4期412-415,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：河北省自然科学基金(H2014405005);河北省教育厅高等学校科学研究计划项目(ZH2012001);河北省中医药管理局科研计划项目(2014202)

摘　　要：目的探讨丹参酮ⅡA对骨髓间充质干细胞(BMSC)定向分化为心肌样细胞的影响。方法取SD大鼠四肢骨骨髓,分离培养BMSC,第2代细胞应用终浓度为500ng/ml丹参酮ⅡA定向诱导(诱导组),将不添加任何诱导剂的BMSC设为对照组。应用免疫细胞化学技术检测结蛋白、α-横纹肌肌动蛋白(α-SCA-actin)和心肌特异性肌钙蛋白T(cTnT)的表达情况。免疫荧光双标染色技术检测α-SCA-actin和cTnT的表达。透射电镜观察2组超微结构的改变。结果免疫细胞化学染色结果显示,与对照组比较,诱导组BMSC中结蛋白(50 279.6±2575.2)vs(1325.6±196.1)、α-SCA-actin(44 738.3±1963.3)vs(1446.3±202.4)、cTnT(27 658.8±2382.2)vs(1369.1±214.2)明显升高(P<0.01)。免疫荧光双标染色法检测诱导组α-SCA-actin阳性为红色,cTnT阳性为绿色,两者重叠部位为黄色,呈现共表达。透射电镜结果显示,诱导组细胞可见平行排列的肌丝并富含线粒体、糖原和核糖体以及丰富的粗面内质网。结论经丹参酮ⅡA诱导的BMSC具有向心肌细胞分化的表型。Objective To study the role of TanshinoneⅡ a in inducing differentiation of BMSC into cardiomyocytes.Methods BMSC isolated from bone marrow of SD rats were cultured and divided into induction group and control group.Expressions ofα-SCA-actin and cTnT were detected with immunofluorescence staining.Ultramicrostructure of BMSC was observed under transmission electron microscope.Results The BMSC were closely arranged in an identical direction,and manifested as spindle and polygonal in shape on day 7after culture,as long spindle in shape on day 10 after culture,and as long cylindrical in shape on day 28 after culture.No morphological change was found in uncultured BMSC.The expression levels and IA value ofα-SCA-actin and cTnT were significantly higher in induction group than in control group（50279.6±2575.2 vs 1325.6±196.1,44 738.3±1963.3 vs 1446.3±202.4,27 658.8±2382.2 vs 1369.1±214.2,P〈0.01）.Immunofluorescence staining showed that theα-SCA-actin was stained red,the cTnT was stained green,their overlapping area was stained yellow.Transmission electron microscopy showed parallel arrangement of myofilaments in BMSC containing rich mitochondria,glycogen,ribosome and rough endoplasmic reticulum.Conclusion TanshinoneⅡ a can induce differentiation of BMSC into cardiomyocytes.

关 键 词：丹参酮 间质干细胞 肌细胞 心脏 结蛋白 肌动蛋白类 肌钙蛋白T 

分 类 号：R542.2[医药卫生—心血管疾病]