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目的探讨5-氮胞苷诱导的大鼠心室成纤维细胞(CFs)植入急性心肌梗死(AMI)区的心肌损伤修复作用。方法取SD大鼠乳鼠心室CFs体外培养、扩增,取第三代细胞加5-氮胞苷10μmol/L诱导,并行5-溴脱氧尿嘧啶核苷(BrdU)标记,收集(0.8～1.2)×106个细胞用于移植。SD大鼠20只,结扎冠状动脉前降支建立大鼠AMI模型,2周后,治疗组(10只)将诱导后的CFs注入梗死区及周边,对照组(10只)予等量不含CFs的培养液接种。6周后处死所有动物,获取心肌标本,HE及免疫组化染色观察CFs分化和转归。结果治疗组HE染色梗死区可见散在带横纹的肌肉组织,电镜下移植细胞及移植细胞与宿主细胞之间均未见闰盘连接;免疫组化染色这些细胞BrdU、β肌球蛋白重链(β-MHC)、肌钙蛋白I(cTnI)及连接蛋白43(Cx43)呈阳性;对照组梗死区为均匀一致的纤维瘢痕组织。结论5-氮胞苷诱导的CFs植入AMI区可分化、转归为带横纹肌细胞,修复心肌损伤。
Objective To investigate the effect of 5-azacytidine-induced myocardial injury in rats with acute myocardial infarction (AMI) implanted with rat cardiac fibroblasts (CFs). Methods SD rat neonatal rat ventricular CFs were cultured and expanded in vitro. The third generation cells were induced with 10μmol / L 5-azacytidine and labeled with BrdU (0.8 ~ 1.2 ) X 106 cells for transplantation. Twenty SD rats were sacrificed and the AMI model was established by ligating the anterior descending coronary artery. After two weeks, the CFs were infused into the infarct area and peripheral area in the treatment group (10 rats) CFs medium inoculation. After 6 weeks, all the animals were sacrificed to obtain the myocardial samples. HE and immunohistochemical staining were used to observe the differentiation and prognosis of CFs. Results In the treatment group, muscle tissue scattered in striated striae was observed in the infarcted area of HE staining. No intercalated disc was found between the transplanted cells and the host cells under electron microscopy. The expression of BrdU and β-myosin heavy chain β-MHC), troponin I (cTnI) and connexin 43 (Cx43) were positive in control group. The control group had homogeneous scar tissue in infarction area. Conclusion 5-azacytidine-induced CFs implanted into AMI can differentiate into striated muscle cells and repair myocardial injury.