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。3d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。
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 marrowmesenchymal 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＆lt;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.
出处
《中国组织工程研究》
CAS
CSCD
2013年第10期1849-1855,共7页
Chinese Journal of Tissue Engineering Research
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
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
