大鼠骨髓单个核细胞移植治疗扩张型心肌病  

Mononuclear bone marrow cells transplantion in treatment of necrotic cardiac muscle in dilated cardiomyopathic rats

作　　者：黄海怡[1] 黄佐[1] 侯健 赵宝珍[3] 周智慧[3] 胡国梁[1] 

机构地区：[1]第二军医大学长征医院心血管内科,上海200003 [2]长征医院血液科 [3]长海医院超声科,上海200433

出　　处：《第二军医大学学报》2004年第9期950-955,共6页Academic Journal of Second Military Medical University

基　　金：上海市科技发展基金 (0 3R1 4 0 2 9)

摘　　要：目的 :研究大鼠骨髓单个核细胞 (m ononuclear bone marrow cells,MBMCs)移植对扩张型心肌病 (dilated cardiomy-opathy,DCM)大鼠心肌修复的作用。 方法 :无菌条件下取成年雄性近交系 Wistar大鼠股骨和胫骨 ,常规抽取骨髓 ,分离制备MBMCs悬液。对 72只成年雄性近交系 Wistar大鼠皮下注射大剂量异丙肾上腺素 (170 mg/ kg,1次 / d,连续 2 d) ,建立 DCM模型。造模成功 (注射后 2周 )后将大鼠随机分成移植组 (n=2 4 )、对照组 (n=2 4 )和假手术组 (n=2 4 ) ,开胸 ,直视下在左心室前壁行心肌注射。移植组注射 4 ',6二乙酰基 - 2 -苯基吲哚 (DAPI)标记的 MBMCs悬液 10 μl(含 5× 10 6个细胞 ) ;对照组注射等量PBS溶液 ;假手术组仅开胸和关胸。于移植后 4周 ,分组进行超声心动图、血流动力学检测 ,并取注射部位心肌组织进行免疫荧光、免疫组化及透射电镜检查。结果 :移植的 MBMCs能在心肌坏死区内存活并形成肌样组织及新生血管 ,结蛋白排列接近正常 ,增殖细胞核抗原 (PCNA)检测阳性 ;透射电镜下可见不成熟的心肌细胞。与对照组和假手术组比较 ,移植组左室收缩和舒张末容积 (L VEDV、L VESV)减小 (P<0 .0 1) ,短轴缩短率 (FS)和射血分数 (EF)增大 (P<0 .0 1) ;左室收缩压 (L VSP)、压力速率最大值及最小值Objective:To investigate the effects of transplanting mononuclear bone marrow cells (MBMCs) in treatment of necrotic cardiac muscle in dilated cardiomyopathic rats. Methods: Subcutaneous injections of isoproterenol(170 mg/kg,once a day for 2 d) were given to 72 male Wistar rats to establish dilated cardiomyopathy(DCM) model. MBMCs were isolated from rats' bone marrow of femurs and tibias and were made into suspension labelled with DAPI before transplantation. Then the labelled cells (5×10 6) and PBS were transplanted into the left ventricular free wall in transplantation and control groups,respectively(n=24).Sham-operated rats(n=24) underwent the surgery but not transplantation. Four weeks after transplantation, the hearts were immunohistochemically processed and the specimens were observed with electron-microscope. Heart function was assessed by echocardiography and Langendorff preparation.Results: The transplanted MBMCs survived in the necrotic myocardium and immunohistochemical examination detected heart muscle actin and neovasculature. Proliferating cell nuclear antigen(PCNA) detecting was positive and the desmin were orderly arranged. Electron microscope revealed immature myocardiocytes. The left ventricular end-diastolic volume (LVEDV) and LV end-systolic volume(LVESV) decreased in transplanting group compared with those in control group and sham-operation group(P<0.01), while the fractional shortening and ejection fraction were increased(P<0.01). The left ventricular systolic pressure(LVSP) and(±dp/dt) also obviously elevated(P<0.01), but the left ventricular end-diastolic pressure(LVEDP) decreased(P<0.01). There were no significant differences between control group and sham-operation group.Conclusion:Transplantation of MBMCs into DCM myocardium can repair necrotic cardiac muscle and improve heart function.

关 键 词：细胞移植 骨髓单个核细胞 扩张型心肌病 心功能 

分 类 号：R542.2[医药卫生—心血管疾病]