电刺激提高分化心肌细胞成熟度并改善心肌梗死大鼠心脏功能

Electrical Stimulation Increases the Maturity of Differentiated Cardiomyocytes and Improves Cardiac Function in Rats with Myocardial Infarction

该研究探讨了电刺激对诱导多能干细胞的心脏分化影响,并评估了分化心肌细胞对心肌梗死的治疗效果。电刺激和非电刺激促进诱导多能干细胞分化出功能性心肌细胞;qRT-PCR和细胞免疫荧光检测分化心肌细胞中心源性基因和功能成熟基因表达情况;建立心肌梗死模型SD大鼠并将其随机分为心肌梗死组、电刺激组和对照组,每组10只。心肌梗死组只结扎冠状动脉,电刺激组和对照组分别在心肌梗死边界注射电刺激和无电刺激预处理的分化心肌细胞。超声心动图和有创血流动力学检测心功能;Masson染色评估梗死面积,免疫组化检测梗死边缘区毛细血管密度。结果表明,电刺激可提高分化心肌细胞的自发搏动,并上调分化心肌细胞中心源性基因(Nkx2-5、GATA4、α-MHC和CX-43)和功能成熟基因(α-actinin和RYR2)的表达(P<0.05)。电刺激明显改善心肌梗死大鼠心功能,减小梗死面积和增加梗死边缘区毛细血管密度(P<0.05)。以上结果表明,电刺激可提高诱导多能干细胞的心脏分化效率并促进分化心肌细胞的成熟;经电刺激预处理的分化心肌细胞可明显改善心肌梗死大鼠心功能。

This study explored the effect of electrical stimulation on the cardiac differentiation of induced pluripotent stem cells and evaluated the therapeutic effect of differentiated cardiomyocytes on myocardial infarction.Electrical stimulation and non-electric stimulation promoted induced pluripotent stem cells to differentiate into functional cardiomyocytes.qRT-PCR and cellular immunofluorescence were used to detect the expression levels of cardiogenic genes and functionally mature genes in differentiated cardiomyocytes.SD rats with myocardial infarction were established and randomly divided into myocardial infarction group,electrical stimulation group and control group,with 10 rats in each group.In the myocardial infarction group,only the coronary arteries were ligated.The electrical stimulation group and the control group were injected with differentiated cardiomyocytes pretreated with electrical stimulation and non-electrical stimulation at the border of myocardial infarction respectively.Echocardiography and invasive hemodynamics were used to detect cardiac function.Masson staining was used to evaluate the infarct area.Immunohistochemistry was used to detect the capillary density in the marginal area of infarction.The results showed that electrical stimulation could increase the spontaneous pulsation of differentiated cardiomyocytes,and up-regulate the expression of cardiogenic genes (Nkx2-5,GATA4,α-MHC and CX-43) and functionally mature genes(α-actinin and RYR2) of differentiated cardiomyocytes (P<0.05).The electrical stimulation significantly improved the cardiac function of rats with myocardial infarction,while reduced the infarct area and increased the capillary density in the marginal area of the infarction (P<0.05).The above results indicated that electrical stimulation could improve the efficiency of cardiac differentiation of induced pluripotent stem cells and promote the maturation of differentiated cardiomyocytes.Differentiated cardiomyocytes pretreated by electrical stimulation could significantly improve the cardiac function of rats with myocardial infarction.