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探讨新生心肌细胞移植重建心脏优势起搏点治疗缓慢性心律失常的可行性。采用酶法分离新生Yorkshire猪右房组织(包括窦房结),获取单个心肌细胞,差速贴壁法纯化,4,6-二脒基-2-苯基吲哚(DAPI)标记后制备成5×106个/ml心肌细胞悬液,开胸直视下将1ml悬液注入细胞移植组幼年猪左室游离壁(n=5),对照组(n=5)注入等体积培养基,移植前3天开始应用环孢素A和泼尼松龙抑制免疫排斥反应。移植3周后应用射频消融技术打断His束造成Ⅲ度房室阻滞。分别应用心电图、Holter、心腔内起搏标测和免疫荧光显像进行电生理学评价和组织学观察。结果:His束消融后,两组动物均形成Ⅲ度房室阻滞。细胞移植组室性节律频率显著快于对照组(移植组平均为95次/分,对照组为32次/分)。心腔内起搏标测证实细胞移植组室性心律起源于细胞移植区。对照组均在24h内发生心室颤动死亡。心脏组织冰冻切片可见DAPI标记的呈蓝色荧光的移植细胞核,移植细胞与宿主心肌细胞间有连接蛋白43和N型钙粘素的表达。结论:移植的心肌细胞可在宿主心脏存活并与周围细胞发生电连接,并主导心室节律。
To investigate the feasibility of neonatal cardiomyocyte transplantation in reconstruction of cardiac superiority pacemaker for the treatment of bradyarrhythmia. The isolated Yorkshire pig right atrial tissues (including sinoatrial node) were isolated by enzymatic method, single cardiomyocytes were obtained and purified by differential adherence and labeled with 4,6-diamidino-2-phenylindole (DAPI) 5 × 106 cells / ml cardiomyocyte suspension was injected into the left porosus free wall (n = 5) and the control group (n = 5) Application of cyclosporine A and prednisolone for the first 3 days suppresses immune rejection. Three weeks after transplantation, radiofrequency ablation was used to break the bundle and cause third-degree atrioventricular block. Electrocardiogram, Holter, intracardiac pacing and immunofluorescence imaging were used for electrophysiological evaluation and histological observation respectively. Results: After beam ablation, both groups of animals formed Ⅲ degree atrioventricular block. The frequency of ventricular rhythm in the cell transplantation group was significantly faster than that in the control group (average 95 / min in the transplantation group and 32 / min in the control group). Intracardiac pacing mapping confirmed cell transplantation group ventricular arrhythmias originated in the cell transplantation area. The control group died of ventricular fibrillation in 24h. Frozen sections of cardiac tissue were observed DAPI labeled blue fluorescence of the transplanted nucleus, transplanted cells and host cardiomyocytes between connexin 43 and N-type cadherin expression. CONCLUSION: Transplanted cardiomyocytes can survive in the host heart and become electrically connected to the surrounding cells and dominate the ventricular rhythm.