中国组织工程研究

中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (10): 1849-1855.doi: 10.3969/j.issn.2095-4344.2013.10.022

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

丹酚酸B及丹参酮ⅡA诱导骨髓间充质干细胞分化为心肌样细胞

李 旭1，王亚玲2   

  1. 1河北北方学院，河北省张家口市 075000
    2河北北方学院附属第一医院心内科，河北省张家口市 075000
  • 收稿日期:2012-05-22 修回日期:2012-07-24 出版日期:2013-03-05 发布日期:2013-03-05
  • 通讯作者: 王亚玲，硕士，副主任医师，河北北方学院附属第一医院心内科，河北省张家口市 075000
  • 作者简介:李旭★，男，1980年生，山西省太原市人，汉族，河北北方学院在读硕士，医师，主要从事心血管疾病的基础与临床研究。lixubfxy@163.com

Salvianolic acid B and tanshinoneⅡA induce bone marrow mesenchymal stem cells to differentiate into cardiomyocyte-like cells

Li Xu1, Wang Ya-ling2   

  1. 1 Hebei North University, Zhangjiakou 075000, Hebei Province, China
    2 Department of Cardiology, First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, Hebei Province, China
  • Received:2012-05-22 Revised:2012-07-24 Online:2013-03-05 Published:2013-03-05
  • Contact: Wang Ya-ling, Master, Department of Cardiology, First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, Hebei Province, China
  • About author:Li Xu★, Studying for master’s degree, Physician, Hebei North University, Zhangjiakou 075000, Hebei Province, China lixubfxy@163.com

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424

被引次数

21

摘要：

背景：目前大多数研究者采用5-氮胞苷诱导骨髓间充质干细胞分化为心肌细胞，存在着一定的不良反应，难以应用于临床。中药丹参本身在临床上广泛用于治疗心血管系统疾病，其主要化学成分为丹酚酸 B和丹参酮ⅡA。
目的：观察丹酚酸B及丹参酮ⅡA联合诱导骨髓间充质干细胞定向分化为心肌样细胞的效果。
方法：取SD大鼠四肢骨骨髓，分离培养骨髓间充质干细胞，应用丹酚酸B、丹参酮ⅡA及二者联合分别对第2代骨髓间充质干细胞定向诱导，不加诱导剂为空白对照组。3 d后各实验组去除诱导培养基，用正常培养基继续培养4周。
结果与结论：空白对照组细胞结蛋白、α-横纹肌肌动蛋白、肌钙蛋白T及缝隙连接蛋白43均为弱阳性或阴性表达。与空白对照组相比，丹酚酸B组，丹参酮ⅡA组，丹酚酸B+丹参酮ⅡA组骨髓间充质干细胞各标记物阳性表达均明显升高，且差异有显著性意义(P < 0.01)。其中，二者联合诱导组各标记物的阳性率均最高。荧光免疫细胞化学鉴定可见诱导组细胞质内结蛋白的表达呈红色，肌钙蛋白T的表达呈绿色，当两者同时被观察时可见其重叠的部位变成黄色。结果显示丹酚酸B及丹参酮ⅡA均可分别诱导骨髓间充质干细胞获得心肌分化表型，且二者联合诱导效果最好。

关键词: 干细胞, 干细胞与中医药, 丹酚酸B, 丹参酮ⅡA, 骨髓间充质干细胞, 心肌细胞, 诱导分化, 结蛋白, 肌动蛋白, 肌钙蛋白, 缝隙连接蛋白43, 干细胞图片文章

Abstract:

BACKGROUND: To date, most researchers use 5-azacytidine to induce the bone marrow mesenchymal stem cells to differentiate into cardiomyocytes, and there are certain adverse reactions which are difficult to be applied in the clinic. Chinese medicine Danshen is widely used in the clinical treatment of cardiovascular diseases, and its main chemical components are salvianolic acid B and tanshinoneⅡA
OBJECTIVE: To explore the effect of salvianolic acid B and tanshinoneⅡA in inducing bone marrow mesenchymal stem cells to differentiate into cardiomyocyte-like cells.
METHODS: Bone marrow mesenchymal stem cells of Sprague-Dawley rats were isolated and cultured from rat bone marrow, and then the second generation of bone marrow mesenchymal stem cells were induced by salvianolic acid B, tanshinoneⅡA and their combination. The cells without inductor were considered as blank control group. After cultured for 3 days, the induction medium was removed and cultured with normal culture medium for 4 weeks.
RESULTS AND CONCLUSION: Desmin, α-sarcomeric actin, cardiac troponin T and connexin 43 in the blank control group showed weakly positive or negative expression. Compared with the blank control group, the positive expressions of desmin, α-sarcomeric actin, cardiac troponin T and connexin 43 in salvianolic acid B group, tanshinoneⅡA group and salvianolic acid B + tanshinoneⅡA group were significantly increased, and the differences were significant (P < 0.01). The positive expression rate of the markers above was highest in the salvianolic acid B + tanshinoneⅡA group. Fluorescent immunocytochemistry staining showed that the expression of desmin in the cells of induction groups was red and the expression of cardiac troponin T was green, while the part that overlaps was yellow. The results showed that salvianolic acid B and tanshinoneⅡA may induce bone marrow mesenchymal stem cells to acquire cardiogenic phenotype separately, and the effect of combined induction is better than single ingredient.

Key words: stem cells, stem cells and traditional Chinese medicine, salvianolic acid B, tanshinoneⅡA, bone marrow mesenchymal stem cells, cardiomyocytes, differentiation, desmin, α-sarcomeric actin, cardiac troponin T, connexin 43, stem cell photographs-containing paper

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