摘要
目的探讨转化生长因子β(TGF-β)对骨髓间充质干细胞(MSCs)定向分化为心肌样细胞的影响。方法取SD大鼠四肢骨髓,分离培养MSCs,应用2、5、10、15 ng/ml TGF-β1对第2代MSCs定向诱导,依次为A组、B组、C组和D组,不加TGF-β1诱导为对照组。各组培养4周后,倒置相差显微镜观察细胞形态学变化,应用免疫细胞化学技术检测结蛋白、α横纹肌肌动蛋白及p38丝裂原活化蛋白激酶(p38MAPK)的表达,通过肌钙蛋白Ⅰ表达计算心肌样细胞的转化率。结果对照组结蛋白、α横纹肌肌动蛋白及p38MAPK均为弱阳性或阴性表达。与对照组比较,其他4组上述各标记物阳性表达均明显升高(P<0.01)。B组表达呈峰值水平,明显高于A、C和D组(P<0.05,P<0.01)。对照组的心肌样细胞转化率较低。与对照组比较,其他4组心肌样细胞转化率均明显升高(P<0.01),B组最高,明显高于A组和D组(P<0.05,P<0.01),但与C组差异无统计学意义(P>0.05)。结论 TGF-β可诱导MSCs获得心肌分化表型,TGF-β1 5 ng/ml可能是一种合适的诱导浓度。
Objective To explore the effect of different concentrations of TGF-β on the differentiation of rat bone marrow MSCs into cardiomyocyte-like cells in vitro. Methods SD rat MSCs were isolated and cultured from rat bone marrow,then the MSCs of 2nd-generation were induced by 2, 5,10,15 ng/ml TGF-β1 for 72 h（group A,B, C and D, respectively）. The cultured cells were observed by phase-contrast microscopy. The immunohistochemical technique was used to determine the expression of desmin, α-sarcomeric actin, cTnI and p38MAPK. Results The morphology of MSCs was different among A,B, C and D groups. MSCs in groups A,D after having induced for lw were same as the control group. MSCs of group B at 4w showed fusiform shape, orientating with one accord, forming myotubule-like structure. The morphology of MSCs of group C was same as that of the group B,but the number of cells was relatively small. MSCs of groups A,B,C and D at 4w could be identified by the positive staining for desmin, α-sareomeric actin and p38MAPK. The positive rates were all higher than those of the control group. The positive rate of cTnI in the group B was higher than that in the group D（P〈0. 01） and group A（P〈0.05）. The cardiomyocyte-like cell transforming rate in the control group was also lower than that in the groups A,B,C and D（P〈0.01）. Conclusion TGF-β may induce MSCs to acquire cardiogenic phenotype and 5 ng/ml may be a suitable inducing concentration.
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2012年第1期62-65,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
