超声微泡破碎技术促进人MxA基因在小鼠肝脏转染的实验研究  

Enhanced human MxA gene expression in mice liver by ultrasound and microbubble destruction technique

作　　者：刘爱平[1] 赵瑞秋[1] 蔡丽君[1] 周方[1] 冉海涛[2] 任红[3] 许红梅[1] 

机构地区：[1]重庆医科大学儿童医院感染消化科,重庆400012 [2]重庆医科大学超声研究所,重庆400016 [3]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《重庆医科大学学报》2010年第3期350-353,共4页Journal of Chongqing Medical University

摘　　要：目的:观察静脉注射基因和微泡声学造影剂,同时经皮超声辐照肝脏的方式是否能增强外源MxA基因在小鼠肝脏中的定位表达,并探讨不同辐照参数对该基因表达的影响。方法:(1)不同转染方法对转染效率的影响:采用昆明小鼠24只,随机分为4组。分别为质粒转染组、微泡+质粒组、超声+质粒组和微泡+超声+质粒组。7d后分别取肝、心、脾、肾、肺和骨骼肌,采用间接免疫荧光法进行MxA蛋白的半定量检测。(2)不同辐照参数对基因表达的影响:采用昆明小鼠20只,随机分为4组。按转染不同超声辐照声强分为0.25W/cm2组、0.5W/cm2组、0.75W/cm2组和1.0W/cm2组。7d后分别取肝组织,采用间接免疫荧光法进行MxA蛋白的半定量检测。结果:(1)肝脏每高倍视野表达阳性细胞数在微泡+超声+质粒组最高,其次为超声+质粒组,再次为微泡+质粒组,表达最少的为质粒组;各组间两两比较,均有显著统计学差异(P<0.001);超声+微泡+质粒组中检测发现,脾组织偶有MxA表达阳性细胞,其余各组仅在肝组织存在MxA表达。(2)肝脏每高倍视野表达阳性细胞数0.5W/cm2组最高,其次为0.75W/cm2组,再次为1.0W/cm2组,0.25W/cm2组表达最少;各组间两两比较,均有显著统计学差异(P<0.001)。结论:超声微泡破碎技术能显著增强外源MxA基因在小鼠肝脏中的靶向性表达;超声辐照频率不变的条件下,在一定范围内增加声强,能促进外源MxA基因在小鼠肝脏表达;当声强为0.5W/cm2时,转染率最高;该技术生物安全性好,为肝脏疾病的基因治疗提供了一项高效、安全的转染技术。Objective：To investigate whether expression of MxA gene could be enhanced by intravenous injection of gene and contrast agent with ultrasound wave irradiation in mice liver and to investigate the effects of different ultrasound parameters on gene expression.Methods：（1） Effect of different methods on transfection efficiency：24 Kunming mice were randomly divided into 4 groups as plasmid group,plasmid and microbubble group,ultrasound and plasmid group,and plasmid,ultrasound and microbubble group.After 7 days,the lungs,hearts,spleens,kidneys,lungs and skeletal muscles were harvested and half quantization of MxA protein was detected with immunofluorescence technique.（2） Effects of different parameters on gene expression：20 Kunming mice were randomly divided into 4 groups as 0.25 W/cm2 group,0.5 W/cm2 group,0.75 W/cm2 group and 1.0 W/cm2 group according to different ultrasound parameters.After 7 days,liver tissues were harvested for the half quantization of MxA by the same method described above.Results：（1） Number of liver cells with positive expression was the highest in plasmid,ultrasound and microbubble group,followed by ultrasound and plasmid group,plasmid and microbubble group,and plasmid group in order;There was significant statistical difference between either two groups（P0.001）.Positive expression of MxA was found in few spleen cells of plasmid,ultrasound and microbubble group while MxA positive expression cells were only found in livers of rest groups.（2） Number of liver cells with positive expression was the highest in 0.5W/cm2 group,followed by 0.75 W/cm2 group,1.0 W/cm2 and 0.25 W/cm2 group in order;There was significant statistical difference between either two groups（P0.001）.Conclusion：Local expression of exogenous MxA gene in mice liver could be significantly enhanced by ultrasound and microbubble destruction technique.In a certain range,expression of human MxA gene in mice liver can be enhanced with increased intensity of sound.Transfection efficiency was the highest

关 键 词：微泡声学造影剂 超声 基因转染 辐照参数 

分 类 号：R454.3[医药卫生—治疗学]