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目的探讨三七皂甙对体外定向诱导猪骨髓基质细胞(MSCs)分化为心肌样细胞的影响。方法用穿刺方法抽取猪骨髓,分离并培养骨髓基质细胞,中药三七皂甙定向诱导分化为心肌样细胞。倒置显微镜下观察细胞形态,免疫细胞化学法鉴定心肌细胞特异性抗原标志物肌钙蛋白T,用RT-PCR对心肌肌球蛋白重链(β-MHC)进行鉴定。取培养的猪心肌细胞作为阳性对照,未经三七皂甙诱导正常培养的骨髓基质细胞作为阴性对照。结果猪MSCs经三七皂甙诱导2h后,大部分MSCs可分化为心肌样细胞,免疫细胞化学检测细胞中的肌钙蛋白T呈阳性,RT-PCR显示能表达β-MHC。结论MSCs是骨髓来源的具有多向分化潜能的干细胞,在三七皂甙的定向诱导下,可以向心肌样细胞分化,有望成为心衰及心肌梗死等心肌损伤时干细胞移植治疗的理想细胞材料。
Objective To investigate the effect of Panax notoginseng saponin on the differentiation of porcine bone marrow stromal cells (MSCs) into cardiomyocyte-like cells in vitro. Methods The porcine bone marrow was extracted by puncture, bone marrow stromal cells were isolated and cultured, and the traditional Chinese medicine Sanqi saponin was induced to differentiate into cardiomyocyte-like cells. Cell morphology was observed under an inverted microscope. Cardiac myocyte-specific antigen troponin T was identified by immunocytochemistry. Myosin heavy chain (β-MHC) was identified by RT-PCR. Cultured porcine myocardium cells were used as positive controls, and normal cultured bone marrow stromal cells without panax notoginseng were used as negative controls. Results After MSCs were induced by panax notoginseng saponin for 2 hours, most MSCs could differentiate into cardiomyocyte-like cells. Immunocytochemistry showed that the troponin T was positive in the cells, and RT-PCR showed that it could express β-MHC. Conclusion MSCs are bone marrow-derived stem cells with multipotent differentiation potential. They can differentiate into cardiomyocyte-like cells under the induction of panax notoginseng saponins, and they are expected to be ideal cell materials for stem cell transplantation in the treatment of heart failure and myocardial infarction.