超声造影剂介导下增强腺病毒相关病毒心肌靶向转染率的实验分析-内科学(心血管)专业论文.docx

新疆医科大学医学博士学位论文 新疆医科大学医学博士学位论文 — — PAGE 6— — —7— The study of enhancement of recombinant adeno-associated virus transgene to myocardium by targeted echo-contrast agent Postgraduate： Yang Shao Ling Supervisor： Mu Yu Ming Abstract Objective: The achievement of organ-specific delivery of gene/drug represents a major problem limiting the clinical application of retroviral vectors for gene therapy, whilst non-viral techniques have the disadvantage of lack of efficiency and longevity of gene expression. Ultrasound-targeted microbubble destruction (UTMD) has been newly developed for destructing the bubbles carrying drugs or genes, and thus for achieving the local release of these target molecules. The aim of this study was to evaluate the potential for achieving to the effective, local delivery of recombinant adeno-associated virus serotypes 2 (rAAV2) transgene into rat myocardium by UTMD. This study involves three parts. The aim of the first part is to study the best titer of rAAV2 delivered to myocardium by targeted ultrasound microbubbles and being infused into tail vein. The aim of the second part is to study the changes of left ventricular function before and after rAAV2-GFP targeted to myocardium by the ultrasound mediated microbubbles. The aim of the third part is to evaluate the potential for achieving to the effective, local delivery of recombinant adeno-associated virus serotype-2 (rAAV2) transgene into rat myocardium by UTMD. Methods: The methods of the fist part: Twenty one adult SD rats were divided into seven groups. SonoVue attached with different titers (1.5×109vg/mL ； 3.0×109vg/mL；1.5×1010 vg/mL；3.0×1010vg/mL; 1.5×1011vg/mL; 3.0×1011vg/mL) of rAAV2-GFP was infused into the tail vein of rats, following ultrasound mediated microbubbles destruction, as experiment groups. Normal saline was infused into the tail vein of rats as the control group (without rAAV2). Rats were killed after 14 days and hearts were harvested. GFP protein expression whic