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目的将携带人超极化激活环核苷酸门控通道2(hHCN2)基因的自体间充质干细胞(MSCs)移植入完全性房室传导阻滞(CHB)猪模型的希氏束中,探讨构建自体生物起搏器的可行性。方法构建包含hHCN2基因的重组腺病毒,转染猪MSCs。构建猪的CHB模型。将转基因的MSCs移植入希氏束后,观察其心率变化及对儿茶酚胺的反应性。对移植部位的心肌行组织学及免疫荧光检查。结果成功构建重组腺病毒pAd.hHCN2并转染猪MSCs。自体移植于CHB动物模型希氏束后,与对照组相比,转基因MSCs显著提升了实验组的心率(P<0.01),且其心律具有儿茶酚胺反应性。移植部位心肌取样检查显示MSCs在心肌中存活并且表达hHCN2蛋白。结论在猪CHB模型中,移植入希氏束的转hHCN2基因的自体MSCs短期内可以发挥生物起搏器的功能。
OBJECTIVE: Transplantation of autologous mesenchymal stem cells (MSCs) carrying human hyperpolarization-activated cyclic nucleotide gate 2 (hHCN2) gene into His bundle of pigs with complete atrioventricular block (CHB) Feasibility of constructing self biological pacemaker. Methods The recombinant adenovirus containing hHCN2 gene was constructed and transfected into porcine MSCs. Construction of pig CHB model. Transgenic MSCs were transplanted into His bundle and observed for changes in heart rate and reactivity to catecholamines. Histological examination and immunofluorescence were performed on the transplanted myocardium. Results The recombinant adenovirus pAd.hHCN2 was successfully constructed and transfected into porcine MSCs. Autologous transplantation in the CHB animal model His bundle, compared with the control group, transgenic MSCs significantly increased the heart rate of the experimental group (P <0.01), and the rhythm of catecholamine reactivity. Cardiac sampling at the transplanted site showed that MSCs survive in the myocardium and express hHCN2 protein. Conclusion In porcine CHB model, autologous MSCs transplanted into His bundle could hinder the function of biological pacemaker in short term.