介入超声靶向投递及辐照增加基因表达安全性研究-第三军医大学学报.DOC

心腔内超声辐照增加基因表达技术的安全性研究 （400038重庆重庆第三军医大学西南医院老年科1；400016重庆2重庆医科大学附属第二医院超声影像研究所） 建立一套包括经皮介入心肌内基因投递、心腔内超声辐照增强基因表达的技术体系，并其安全性。方法 研制心腔内超声辐照导管（ntracardiac Ultrasound，US），经常规介入法置于活体犬左心室，并经导管中特制的微型针刺入心肌注射报告基因绿色荧光蛋白（Enhanced Green Fluorecence ProteinEGFP）质粒健康杂种犬12只，体15～20 kg，雌雄不，分为EGFP+US组、单纯EGFP组。在EGFP+US组，注射质粒500 μg后，心腔内超声辐照1（1-MHz、1-W/cm2、60s）。单纯EGFP组仅注射质粒，继续饲养犬，测定照射前、照射过程中、照射后心肌酶谱变化，7后，测定心肌、肝、肾、肺脏、肌肉中EGFP mRNA及蛋白质表达，观察心肌及上述脏器的病理形态变化。结果 经超声导管成功实现了经皮心肌内注入EGFP质粒，心肌形态正常，心腔内超声辐照后明显增加EGFP基因在心肌表达，与单纯EGFP组相比，EGFP+US组EGFPmRNA表达增加6.5倍，蛋白表达增加4.1倍(P0.01)。心肌无损伤、出血，辐照前后心肌酶谱没有变化(P＞0.05)。光镜下肺脏、肝脏、肾脏形态无改变，也未见EGFP表达心肌外未见质粒沉积。结论 心腔内超声辐照裸质粒在活体心肌组织中转染表达超声辐照对心脏毒副影响，心腔内超声辐照增强基因表达安全。 关键词心腔内超声导管基因表达安全性 ultrasound irradiation increases gene expression: A study of its safety Qiao yingyuan1， Jing uang2， Liang yi1， Xing shengi2（1Department of Geriatrics,Southwest Hospital,Third Military Medical University, Chongqing 400038, China; 2Institute of Ultrasound Imaging, Second Affiliated Hospital of Chongqing Medical University，Chongqing 400010，China [Abstract] Objective To establish the safety of a novel ultrasound catheter system that can increase gene expression by delivering transendocardial and intracardiac ultrasound irradiation. Methods An intracardiac ultrasound (US)irradiation catheter was placed in the left ventricle of living dogs. Enhanced green fluorecence protein plasmids (EGFP) were injected into myocardium through a special miniature needle in the catheter. Twelve health hybrid dogs at the age of 12 months, weighing 15-20 kg, were divided into EGFP+US group and EGFP group. Dogs in EGFP+US group were injected with 500 μg EGFP and then underwent intracardiac US irradiation for 1 min (1-MHz，1-W/cm2 EGFP group were injected with the same amount of EGFP.. CK secretion was measured before，during . mRNA and protein expressions of EGFP in myocardium, liver, kidney and lung were detected on day 7 after irradiation to observe changes in their pathological morphology. Results EGFP were succes