摘要
目的观察养心通脉有效部位方对大鼠骨髓间充质干细胞(MSCs)体外分化为心肌样细胞的诱导作用。方法体外培养大鼠MSCs,设养心通脉有效部位方(YTF)组、5-氮胞苷(5-aza)阳性对照组和低糖DMEM组,体外诱导分化心肌样细胞;用免疫细胞化学方法观测其心肌肌丝蛋白(Desmin)、肌球蛋白重链(MHC)的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达。结果大鼠MSCs体外向心肌样细胞分化诱导后,在DMEM组中未发现Desmin和MHC阳性细胞;而YTF组和5-aza组的Desmin及MHC免疫组化染色均可见阳性免疫复合物,与DMEM组比较,YTF组和5-aza组Desmin、MHC、GATA-4 mRNA表达的相对光密度(IOD)值均明显提高(P<0.01,P<0.05),而且YTF组Desmin、MHC的IOD也明显高于5-aza组(P<0.05),但YTF组GATA-4 mRNA的IOD值与5-aza组比较无统计学差异。结论 YTF在体外可诱导MSCs向心肌样细胞分化。
Objective To observe the impact of effective fractions of Yangxin Tongmai Formula（YTF） on cardiomyo- genic differentiation of rat bone marrow mesenchymal stem cells（MSCs） in vitro. Methods MSCs were cultured in vit- ro, and then divided into three groups： YTF effective fractions group（YFT group）, 5-azacytidine group（5-aza group, positive control group） and low glucose DMEM cell culture medium group（DMEM group, negative control group）. MSCs were applied to induce cardiomyogenic differentiation in vitro. The expression of myofilament protein （desmin） and myosin heavy chain （MHC） was detected by immunocytochemistry assay, and the mRNA expression of myocardial transcription factor GATA-4 was detected by RT-PCR. Results After in-vitro eardiomyogenic differentia- tion induction in MSCs of three groups, desmin and MHC positive cells were not found in DMEM group, but in YTF group and 5-aza group, desmin and MHC positive immunocomplex was found by immunohistochemieal assay. More- over, we found that the IOD of higher than that higher than that in DMEM group in 5-aza group desmin, MHC and GATA-4 mRNA in YTF group and 5-aza group was significantly （P 〈 0.01 or P 〈 0.05）, the IOD of desmin and MHC in YTF group was significantly （P 〈 0.05）, and there was no significantly difference between YTF-group and 5-aza group in the IOD of GATA-4 mRNA. Conclusion Yangxin Tongmai Formula effective fractions can induce the car- diomyogenic differentiation of rat bone marrow mesenchymal stem cells in vitro.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2012年第4期427-430,共4页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金(30772696,30973717)
湖南省研究生创新基金(CX2009B168)
关键词
骨髓间充质干细胞
心肌样细胞
分化
养心通脉有效部位方
Bone marrow mesenchymal stem cells
Cardiomyocyte-like cells
Differentiation
Yaugxiu Tougmai Formula effective fractions
