淫羊藿苷对经血源性子宫内膜干细胞成心肌样细胞分化的影响

Icariin-Mediated Differentiation of Menstrual Blood-Derived Endometrial Stem Cells into Cardiomyocyte-Like Cells

选择安全高效的诱导剂促进干细胞成心肌样细胞分化,将为干细胞治疗心脏疾病提供更佳的种子细胞,加强干细胞治疗效果。该研究重在明确淫羊藿苷(icariin,ICA)对经血源性子宫内膜干细胞(menstrual blood-derived endometrial stem cells,Men ESC)成心肌样细胞分化的影响。首先,通过MTT法和划痕实验检测ICA对Men ESC增殖和迁移的影响。RT-PCR检测ICA与5氮杂胞苷(5azacytidine,5aza C)诱导前后心肌特异性转录调控因子肌细胞增强因子-2C(myocyte enhancer factor-2C,MEF-2C)、NK2转录因子相关蛋白5(NK2 transcription factor related locus 5,Nkx2.5)、GATA结合蛋白4(GATA binding protein 4,GATA4)以及心肌特异性标志物人类心房钠尿肽(human atrial natriuretic peptide,h ANP)、肌钙蛋白I(cardiac tropomin I,c Tn I)、肌钙蛋白T(cardiac tropomin T,c Tn T)基因的表达水平。免疫荧光检测ICA与5aza C诱导后细胞中c Tn T的蛋白质水平表达。结果表明,ICA对Men ESC的增殖和迁移无显著性影响;进一步发现,ICA与5aza C诱导Men ESC后,均能显著上调MEF-2C、Nkx2.5、GATA4、h ANP、c Tn I及c Tn T基因的表达;而最终的免疫荧光结果也证实,ICA诱导后的Men ESC阳性表达c Tn T。综上所述,ICA可在不影响Men ESC增殖和迁移活性的基础上促进其成心肌样细胞分化,为Men ESC在临床上治疗心脏疾病提供新途径。

Application of safe agents to effectively predifferentiate stem cells into cardiomyocyte-like cells would strength the stem cell-based therapies for heart diseases. The aim of this study is to examine the effect of icariin （ICA） on the cardiogenic differentiation of menstrual blood-derived endometrial stem cells （MenESC）. MTT and wound healing assays are used to investigate the effect of ICA on the proliferation and migration of MertESC. Furthermore, RT-PCR and specific fluorescence staining are performed to examine the expression of cardiac-specific transcription factors myocyte enhancer factor-2C （MEF-2C）, NK2 transcription factor related locus 5 （Nkx2.5） and GATA binding protein 4 （GATA4） and cardiac-specific marker human atrial natriuretic peptide （hANP）, cardiac tropomin I （cTnI） and cardiac tropomin T （cTnT） in MenESC treated with ICA or/and 5azacytidine （5azaC）. Our results demonstrated that ICA had no negative effect on the proliferation and migration of MenESC. Icariin and 5azaC treatment could significantly up-regulate the gene expression ofMEF-2C, Nkx2.5, GATA4, hANP, cTnI and cTnT in MenESC. Meanwhile, cardiogenic differentiation was also confirmed by the positive expression of cTnT in MenESC treated with ICA and 5azaC. These results confirmed the safety and effects of cardiogenic differentiation of MenESC treated with ICA, and provided support for the stem cell-based therapies for heart diseases.