【摘要】 目的观察重组人促红细胞生成素(rHu-EPO)联合自体外周血干细胞移植对猪急性心肌梗死(AMI)后心肌病理学和细胞凋亡的影响。方法中华小型猪30只,均采用经皮穿刺球囊封堵冠状动脉左前降支制备猪AMI模型,造模过程中死亡2只,建模成功后随机分为3组。移植组(n=10)建模后1h给予粒细胞集落刺激因子(G-CSF),连用7d,建模第8天采用血细胞分离机单采外周血单个核细胞(PBMC)悬液,经5-溴脱氧尿嘧啶核苷标记,通过经皮经腔导管技术建立的梗死相关动脉通道回输PBMC。实验组(n=9)建模后1h给予G-CSF,连用7d,rHu-EPO连用5d,并于建模后第8天以相同方法采集、标记、回输PBMC。对照组(n=9)建模后仅给予等量生理盐水。移植3周后处死动物,制作心肌组织苏木精-伊红染色切片行病理学检查;原位末端标记法染色计算心肌细胞凋亡指数;Western blot法检测各组心肌组织中Bax、Bcl-2及Caspase-3蛋白的表达。结果苏木精-伊红染色显示,对照组梗死区心肌细胞坏死崩解、细胞核消失;移植组大量炎性细胞及成纤维细胞浸润,其间有部分存活心肌细胞;实验组可见大量存活心肌细胞。实验组心肌细胞凋亡指数显著低于移植组和对照组(F=6.491,P<0.05)。与对照组相比,实验组、移植组Bcl-2蛋白表达显著增高,Bax及Caspase-3蛋白表达显著降低;与移植组相比,实验组Bcl-2及Bax蛋白表达显著增高(F=6.513～8.267,P<0.05)。结论 rHu-EPO联合自体外周血干细胞移植治疗AMI可减轻心肌病理学改变,抑制心肌细胞凋亡,其作用可能是通过下调凋亡蛋白、上调抗凋亡蛋白的表达实现的。更多还原

【Abstract】 Objective To observe the effects of recombinant human erythropoietin(rhEPO) combined with autologous peripheral blood stem cells transplantation on pathology and apoptosis in pig after myocardial infarction.Methods A model ofacute myocardial infraction(AMI) was created in 30 minipigs by balloon occlusion of left anterior descending branch of coronary artery,two died at the procedures,the rest were divided into transplantation(n=10),experimental(n=9) and control(n=9) groups.In transplantation group,granulocyte-colony stimulating factor(G-CSF) was given(one hour after completion of the model) for seven days.On day 8,peripheral blood mononuclear cell(PBMC) suspensions was marked with 5-deoxyuridine,the PBMC was then transfused back by way of infarction-related artery created by percutaneous catheterization;In experimental group,G-CSF was offered for seven days,and rhEPO for five days starting at one hour after completion of the model.On day 8,PBMC was transfused back with the same method as done in the transplantation group;In control group,only equal volume of normal saline was given.The animals were sacrificed three weeks after transplantation,a slice of myocardium stained with HE was made for pathologic examination;TUNEL was use to calculate the index of apoptosis;Bcl-2,Bax and Caspase-3 protein were tested by Western blot.Results Pathologically,cardiac muscle cell necrosis and disintegration,and disappearance of cell nucleus in infracted areas were observed;In transplantation group,a great quantity of inflammatory cells and fibroblasts were seen,of which,there were some survival cardiac myocytes;In experimental group,mass survival cardiac myocytes could be seen.The apoptotic index in experimental group was lower than that in the transplantation and control groups(F=6.491,P<0.05).Compared with the control,the expressions of Bcl-2 in experimental and transplantation groups were higher,and that of Bax and Caspase-3 were lower;Compared with transplantation group,the Bcl-2 and Bax expressions in experimental group were higher(F=6.513-8.267,P<0.05).Conclusion rhEPO combined with APBMC transplan-tation in the treatment of AMI can relieve pathologic changes and inhibit apoptosis of cardiac myocytes possibly through down-regulating Caspase-3 and Bax and up-regulating Bcl-2 expressions.更多还原