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目的观察心肌内和冠状动脉内注射法移植自体骨骼肌成肌细胞于急性心肌梗死区的生长分化特点。方法以改良成年犬骨骼肌成肌细胞培养方法进行细胞分离及扩增。结扎冠状动脉前降支中段,建立急性心肌梗死模型。分为直接注射对照、移植,冠状动脉内注射对照、移植4组,每组5只。不开通冠状动脉,分别向梗死心肌内和梗死相关冠状动脉内注射自体骨骼肌成肌细胞(1·0~1·4×108个)或等量生理盐水。移植4周后通过HE染色、PTH染色、骨骼肌特异性慢肌球蛋白抗体免疫组化染色和透射电镜寻找梗死区内存在新生肌组织的证据并观察其生长特点。结果经心肌内直接注射和冠状动脉内注射移植自体骨骼肌成肌细胞4周后,透射电镜及HE染色下均可在梗死区内找到新生幼稚肌原性细胞存在,PTH染色证实有新生的横纹肌组织形成,骨骼肌特异性慢肌球蛋白抗体免疫组化染色发现有骨骼肌原性的成熟肌组织存在;成肌细胞直接注射组内新生的肌组织排列较为密集,而冠状动脉内成肌细胞注射组内的新生肌组织排列较分散。结论通过心肌内直接注射和经梗死相关冠状动脉注射将自体骨骼肌成肌细胞移植到急性心肌梗死区后均能形成成熟的肌组织,为以骨骼肌成肌细胞进行急性心肌梗死的细胞心肌成形治疗提供了组织学依据。
OBJECTIVE: To observe the growth and differentiation of autologous skeletal myoblasts transplanted into myocardium by intramyocardial and intracoronary injection in acute myocardial infarction area. Methods The cells of adult canine skeletal muscle myoblasts were isolated and expanded. Ligation of the middle part of the anterior descending coronary artery, the establishment of acute myocardial infarction model. Divided into direct injection control, transplantation, intracoronary injection control, transplantation 4 groups, each group of 5. Without opening the coronary artery, autologous skeletal muscle myoblasts (1.0 ~ 1.4 × 108) or normal saline were injected into the infarct-related myocardium respectively. Four weeks after transplantation, the evidence of new myoblasts in the infarct area and the growth characteristics were observed by HE staining, PTH staining, skeletal muscle-specific slow myosin antibody immunohistochemistry and transmission electron microscopy. Results After 4 weeks of autologous skeletal myoblasts transplantation by intramyocardial injection and intracoronary injection, the appearance of naive myogenic myoblasts could be found in the infarct zone by transmission electron microscopy and HE staining. PTH staining showed that there were newborn striated muscle Tissue formation and skeletal muscle specific anti-myosin antibody immunohistochemical staining found skeletal myogenic mature muscle tissue; myofibroblasts direct injection of newborn muscle tissue arranged more dense, and coronary artery myoblast The newborn muscle tissue within the injection group is more scattered. Conclusions Autologous skeletal myoblasts can be induced to mature muscle tissue by intramyocardial direct injection and infarction-related coronary injection, which is a kind of cardiac myocyte formation in acute myocardial infarction with skeletal muscle myoblasts The treatment provided a histological basis.