钙离子ATP酶2a基因修饰的骨髓干细胞移植治疗大鼠慢性心力衰竭  

Sarcoplasmic reticulum Ca^(2+)-ATPase gene modified mesenchymal stem cell transplantation for treatment of chronic cardiac failure in rats

作　　者：郭豫涛[1] 李小鹰[1] 鲁小春[1] 吴迪[2] 姚克群[2] 陈平[3] 马康涛[3] 周春燕[3] 

机构地区：[1]解放军总医院老年心血管病一科,北京市100853 [2]解放军空军总医院超声科,北京市100036 [3]北京大学医学部生物化学系,北京市100083

出　　处：《中国组织工程研究与临床康复》2007年第7期1267-1270,I0004,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家重大基础研究发展规划九七三计划资助项目(G2000056906)~~

摘　　要：目的:观察肌浆网钙离子ATP酶2a基因修饰的骨髓干细胞移植对慢性心力衰竭大鼠心功能的影响。方法:实验于2004-07/2005-12在北京医科大学生物化学系实验室完成。选取4周龄清洁级雄性SD大鼠作为骨髓供体。雌性SD大鼠作为细胞移植、基因治疗受体,随机数字表法分为4组:肌浆网钙离子ATP酶2a基因治疗组(n=7)、骨髓干细胞移植治疗组(n=7)、肌浆网钙离子ATP酶2a基因修饰的骨髓干细胞移植组(n=8)、腺病毒空载体对照组(n=7)。结扎左冠状动脉制作急性心肌梗死后慢性心力衰竭大鼠模型,每组进行相应的基因治疗和细胞移植。治疗后21d,采用苏木精-伊红染色和MASSON染色评价心脏形态及结构变化,组织多普勒评价各组心功能。并检测肌浆网钙离子ATP酶2a基因和蛋白在宿主心脏的表达。结果:29只雌性大鼠均进入结果分析。①与腺病毒空载体对照组大鼠相比,骨髓干细胞移植治疗组和肌浆网钙离子ATP酶2a基因修饰的骨髓干细胞移植治疗组大鼠心室腔明显减小。MASSON染色显示,肌浆网钙离子ATP酶2a基因修饰的骨髓干细胞移植治疗组大鼠蓝色的胶原纤维染色区域减少,红色的肌纤维染色区域增多。②肌浆网钙离子ATP酶2a基因治疗组、骨髓干细胞移植治疗组和肌浆网钙离子ATP酶2a基因修饰的骨髓干细胞移植治疗组大鼠左室前壁、室间隔收缩及舒张期纵向峰值速度均较腺病毒空载体对照组显著升高[左室前壁:(0.58±0.03),(0.67±0.02),(0.78±0.05),(0.31±0.02)cm/s;(0.81±0.04),(1.26±0.04),(1.39±0.05),(0.40±0.01)cm/s;室间隔:(0.60±0.05),(1.00±0.08),(1.33±0.04),(0.40±0.01)cm/s;(0.70±0.04),(1.28±0.05),(1.57±0.03),(0.44±0.03)cm/s,P<0.001]。与肌浆网钙离子ATP酶2a基因治疗组相比,肌浆网钙离子ATP酶2a基因修饰的骨髓干细胞移植治疗组大鼠左室前壁、室间隔收缩及舒张期纵向峰值速度升高(P<0.001)。③肌浆网钙离子ATP酶2a基因治�AIM： To observe the effect of bone marrow mesenchymal stem cell （MSC） modified with sarcoplasmic reticulum Ca^2＋-ATPase （SERCa2a） gene therapy on cardiac function of rats with chronic heart failure （CCF）. METHODS： The experiment was conducted in the Laboratory of Biochemistry, Capital Medical University between July 2004 and December 2005. MSC was obtained from 4-week male SD rats of clean grade, and female SD rats were selected to be the recipients of cellular transplant and gene therapy, which were randomly divided into 4 groups： SERCa2a gene treatment group （n =7）, MSC transplantation group （n =7） SERCa2a gene modified MSC transplantation group （n =8） and empty adenoviral vector control group （n =7）. Rats were made into models of CCF after acute myocardial infaction by ligation of the left coronary artery, and then rats in each group underwent corresponding gene therapy and cellular transplantation. Twenty-one days later, HE staining and MASSON staining were adopted to evaluate the changes in cardiac morphous and structure, and the cardiac function was evaluated by Doppler tissue imaging （DTI）. SERCa2a gene and protein expressions were inspected respectively. RESULTS： All the 29 female rats ware involved in the analysis of results. （1） Compared to empty adenoviral venctor control group, the ventricular chamber was significantly reduced in the MSC transplantation group and SERCa2a gene modified MSC transplantation group. MASSON staining suggested that stained region of blue collagen fiber in rats was less in the SERCa2a gene modified MSC transplantation group, while stained region of red muscle fiber increased. （2） The peak systolic and diastolic velocity of left ventricular anterior wall and interventricular septum in SERCa2a gene treatment group, MSC transplantation group and SERCa2a gene modified MSC transplantation group were significantly higher than those in the empty adenoviral vector control group [Left ventricular anterior wall： （0.58±0.03）,（0.

关 键 词：细胞移植 干细胞 基因疗法 心力衰竭 充血性 肌浆网 Ca(2+)转运ATP酶/代谢 

分 类 号：R394.2[医药卫生—医学遗传学]