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目的探讨Apelin对人诱导性多能干细胞(hiPSC)的心肌定向分化作用。方法 hiPSC经悬浮法诱导形成拟胚体,分为Apelin处理组(Apelin组)和对照组。Apelin组在高糖DMEM培养液中加入10μmol/ml Apelin,分别于第3、7、11、15、21天采用RT-PCR、Western blot法检测及免疫荧光染色,观察干细胞全能性标记物Oct4、心肌祖细胞标记物Nkx 2.5、心肌特异性肌钙蛋白I(cTnI)的表达变化。结果 Apelin组iPSC在诱导分化后第3天Oct4和Nanog基因表达水平明显降低,分别为同期对照组的(63.2±10.1)%和(51.7±1.6)%(P<0.01);诱导分化后第7天Nkx 2.5和cTnI基因表达水平明显升高,分别为同期对照组的(162.4±22.5)%和(250.6±22.3)%(P<0.05,P<0.01);诱导分化后第15天时,cTnI表达为同期对照组的(156.2±6.4)%(P<0.05);免疫荧光染色显示,cTnI阳性细胞明显多于对照组。结论 Apelin可明显促进hiPSC的心肌定向分化。
Objective To investigate the effect of Apelin on the cardiac myogenic differentiation of human induced pluripotent stem cells (hiPSCs). Methods hiPSCs were induced to form embryoid bodies by suspension method and divided into Apelin group and Apelin group. The Apelin group was treated with 10 μmol / ml Apelin in high glucose DMEM medium, and detected by RT-PCR, Western blot and immunofluorescence staining on days 3, 7, 11, 15, and 21 respectively. Oct4, Cardiac progenitor cell marker Nkx 2.5, cardiac troponin I (cTnI) expression changes. Results The expressions of Oct4 and Nanog in the ipepticeps of Apelin group were significantly decreased (63.2 ± 10.1% vs. 51.7 ± 1.6%, P <0.01) on the 3rd day after differentiation, respectively The expression of cTnI and Nkx2.5 mRNA and cTnI gene were significantly higher in the control group (162.4 ± 22.5% vs 250.6 ± 22.3%, P <0.05, P <0.01, respectively) (156.2 ± 6.4)% (P <0.05) in the control group. Immunofluorescence staining showed that there were more cTnI positive cells than the control group. Conclusion Apelin can obviously promote the directional differentiation of hiPSCs.