经腹心包腔内注射术在慢性心力衰竭大鼠基因治疗中的应用  

The value of intrapericardial injection with a trans-diaphrag matic approach in the gene therapy of chronic heart failure rats

作　　者：惠海鹏[1] 李小鹰[1] 刘涛[2] 刘秀华[3] 孙胜[3] 

机构地区：[1]中国人民解放军总医院老年心内科,北京100853 [2]中国人民解放军总医院老年超声科,北京100853 [3]中国人民解放军总医院病理生理研究室,北京100853

出　　处：《中国病理生理杂志》2005年第4期674-678,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目 (No .30 4 70 716 ) ;国家重点基础研究发展规划 (973) (No .G2 0 0 0 0 5 6 90 6 )

摘　　要：目的:探讨在慢性心力衰竭大鼠中经腹心包腔内注射术的可行性、安全性和有效性及其在心脏疾病基因治疗中的应用价值;探讨SERCA2a基因转导对慢性心力衰竭的治疗作用。方法:采用腹主动脉缩窄术制备慢性心力衰竭模型。应用经腹心包腔内注射术,分别将携带绿色荧光蛋白(eGFP)基因的重组腺相关病毒和携带心肌肌浆网Ca2 + -ATPase(SERCA2a)基因的重组腺相关病毒转导入HF +EGFP组和HF +SERCA2a组大鼠心包腔内,30d后检测各组大鼠的血流动力学指标,荧光显微镜下检查绿色荧光蛋白表达情况和用Westernblot检测心肌肌浆网Ca2 +-ATPase蛋白的表达情况。结果:HF +EGFP组大鼠心肌组织在荧光显微镜下发出弥漫的绿色荧光。HF +SERCA2a组大鼠心肌表达心肌肌浆网Ca2 + -ATPase蛋白明显高于HF组和HF +EGFP组大鼠,因而,其心功能也较另外2组大鼠明显改善,达到正常大鼠的水平。结论:经腹心包腔内注射术为慢性心力衰竭的基因治疗提供了一种简单、有效、安全、经济的方法。SERCA2a基因转导为心力衰竭的治疗提供了一个新的思路。AIM: To study the feasibility, security a nd validity of the intrapericardial injection with a trans-diaphragmatic approach in chronic heart failure rats and its value in the study of gene therapy for hea rt diseases, and further investigate whether adeno-associated virual gene transf er of sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) gene can improve ven tricular function in chronic heart failure (HF) rats. METHODS: An animal model of heart failure was obtained by creati ng descending aortic constriction in rats. Recombinant adeno-associated virus, c arrying enhanced green fluorescent protein (EGFP) gene and recombinant adeno-ass ociated virus carrying SERCA2a gene, were respectively injected into pericardium of heart failure rats in different groups (group HF+EGFP and group HF+SERCA2a) by intrapericardial injection with a trans-diaphragmatic approach. After 30 days , hemodynamic parameters were measured and analyzed. Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein, and Western blotting was performed to detect the expression of SERCA2a. RESULTS: Green fluorescence was detected in cryosection of the h earts in all rats in group HF+EGFP and the expression of green fluorescent prote in was ubiquitously. The expression of SERCA2a in all rats in group HF+SERCA2a w as more than those in group HF and group HF+EGFP. And overexpression of SERCA2a improved the systolic and diastolic function of heart failure rats significantly and the hemodynamic parameters were similar with those of the controls. CONCLUSION: The intrapericardial injection with a trans-diaphrag matic approach suggests a simple, safe, efficient and cheap technique for the ge ne therapy of chronic heart failure. Gene thransfer of SERCA2a may be a new appr oach for the treatment of chronic heart failure.

关 键 词：慢性心力衰竭 心包腔 手术 基因治疗 肌浆网Ca^2+-ATPase 

分 类 号：R363[医药卫生—病理学]