钙离子ATP酶2a基因修饰的骨髓干细胞移植治疗大鼠心肌梗死后心力衰竭的研究  

Bone marrow stem cells-based SERCA2a gene therapy for heart failure after acute myocardium infarction

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,北京100853 [2]空军总医院超声科 [3]北京大学医学部生物化学系

出　　处：《中华医学杂志》2006年第12期826-831,共6页National Medical Journal of China

基　　金：国家"973"重点基础研究发展计划基金资助项目(G2000056906)

摘　　要：目的比较肌浆网钙离子ATP酶2 a(SERCA2 a)基因治疗,骨髓干细胞(MSC)移植以及SERCA2 a基因修饰的细胞移植治疗慢性缺血性心力衰竭的效应,评价MSC作为治疗基因的细胞载体策略的可行性。方法制作心力衰竭大鼠模型并随机分为4组。SERCA2 a基因治疗组(组Ⅰ,7只),MSC细胞移植组(组Ⅱ,7只),基因修饰的细胞移植组(组Ⅲ,8只),腺病毒空载体对照组(组Ⅳ,7只)。检测各组SERCA2 a基因,蛋白表达及活性。超声心动图及有创血流动力学评价心脏功能。HE染色观察心室形态。结果组Ⅰ,组Ⅱ,组Ⅲ比对照组Ⅳ心功能和血流动力学明显改善。治疗后组Ⅱ(2.9 mm±0.2 mm)和组Ⅲ(3.0 mm±0.1 mm)大鼠室壁厚度增加,心室腔减小。治疗后14d组Ⅰ,组Ⅱ与组Ⅲ射血分数(EF)为26.6%±3.9%,20.6%±5.0%,25.6%±2.7%。短轴缩短率(FS)分别为13.3%±2.0%,10.1%±2.5%,12.1%±1.0%(P<0.05)。治疗后21 d组Ⅰ,组Ⅱ与组ⅢEF分别为19.7%±5.0%,21.4%±5.2%,26.6%±4.1%,FS分别为9.7%±2.5%,10.7%±2.6%,12.8%±2.2%(P<0.05)。组Ⅳ治疗后14 d EF无变化,治疗后21 d EF下降了6.6%±4.0%。组Ⅰ和组ⅢSERCA2 a基因,蛋白表达及酶活性均高于组Ⅱ和组Ⅳ(P<0.01)。结论MSC移植和SERCA2 a基因修饰的细胞移植组大鼠呈现稳定持续的心功能改善作用。MSC可以作为治疗基因的有效转移载体。Objective Explore the possibility of MSC to be used to target delivery of therapeutic gene and evaluate the therapeutic effects among gene therapy, MSC transplantation and MSC-based gene therapy. Methods MSC were infected with an adenoviral expression vector carrying SERCA2a. SD female rats were used to make animal model with heart failure after AMI and divided into 4 groups randomly. Group Ⅰ （ n = 7） received SERCA2a gene therapy, group Ⅱ （ n = 7 ） received MSC transplantation, group Ⅲ （ n = 8） received MSC infected with SERCA2a gene transplantation, and group Ⅳ （n = 7 ） received empty adenoviral vector. Cardiac function was evaluated by echocardiography and physiological recorder. SERCA2a gene and protein expression were evaluated by RT-PCR and Western blot respectively. Results Compared to group Ⅳ, EF and FS of group Ⅰ , group Ⅱ and group m were elevated significantly on 14 days after therapy （EF： 67.7 ±3.9, 62.6 ±4.0, 67.9 ±3.7 versus 45.0 ±2.2; FS： 33.9 ± 1.9,31.1 ±2.0, 33.9 ± 1.9 versus 22. 5 ± 1.1, P 〈 0. 05）. While the elevation values of EF and FS began to reduce in group Ⅰ 14 days after, it continued to increase in both group Ⅱ and group Ⅲ. Absolute value of LVEDP at 21 days after treatment was increased in group Ⅰ , group Ⅱ and group Ⅲ compared to group Ⅳ （5. 3 mm Hg ± 1.2 mm Hg,6. 0 ±1.3 mm Hg,6. 2 mm Hg±1. 2 mm Hg versus 1.5 mm Hg ±0. 2 mm Hg, P〈0.05）, as well as absolute value of DP/dtmin （4756 mm Hg/s ±270 mm Hg/s, 5028 mm Hg/s ±253 mm Hg/s, 5283 mm Hg/s±363 mm Hg/s versus 3201 mmHg/s ±211 mm Hg/s, P 〈0.05）. DP/dtmax at 21 days after treatment increased in group Ⅰ , group Ⅱ and group Ⅲ compared to group Ⅳ （6026 mm Hg/s ± 281 mm Hg/s, 6278 mm Hg/s ±319 mm Hg/s, 7057 mmHg/s ±389 mm Hg/s versus 5293 mm Hg/s ±360 mm Hg/s, P 〈 0. 05 ）. SERCA2a expressions and enzyme activity were significantly stronger in group Ⅰ and group Ⅲ than in group Ⅱ and group Ⅳ. Conclusion It showed that all MSC transplant

关 键 词：钙离子 ATP酶2a 基因修饰 骨髓干细胞移植 治疗 大鼠 心肌梗死 心力衰竭 

分 类 号：R542.22[医药卫生—心血管疾病] R541.6[医药卫生—内科学]