| 冉海涛,任红,王志刚,郑元义,张群霞,李小东,许川山.超声波与微泡声学造影剂增强基因定位转染动物实验研究[J].中国医学影像技术,2005,21(8):1158~1160 |
| 超声波与微泡声学造影剂增强基因定位转染动物实验研究 |
| Enhanced gene delivery by ultrasound and contrast agent: experimental study |
| 投稿时间：2005-07-06 修订日期：2005-07-25 |
| DOI： |
| 中文关键词: 超声造影剂,微泡 基因转染 |
| 英文关键词:Ultrasound contrast agent, microbubbles Gene transfection |
| 基金项目:本研究受国家自然科学基金重点项目资助(30430230)。 |
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| 中文摘要: |
| 目的 观察经静脉注射基因与微泡声学造影剂,同时经皮超声辐照肝脏方式能否增强小鼠肝脏基因定位转染及其转染效率。方法 昆明种小白鼠24只,随机分为4组,每组6只。第1组为单纯质粒转染组:经尾静脉快速注入2 ml含20 μg乙型肝炎核心抗原(pcDNA3.1/HBV)质粒的生理盐水溶液。第2组为单纯微泡转染组:经尾静脉快速注入2 ml含20 μg质粒的微泡声学造影剂。第3组为单纯超声转染组:经尾静脉快速注入2 ml含20 μg质粒的生理盐水溶液,同时采用频率为1 MHz、声强为0.5 W/cm2的超声波经小白鼠体表肝区辐照1 min。第4组为超声与微泡转染组:经尾静脉快速注入2 ml含20 μg质粒的微泡声学造影剂,同时经小白鼠体表肝区局部采用相同剂量超声辐照1 min。7天后处死小鼠,分别取肝、肾、肺、心、脾、骨骼肌组织进行pcDNA3.1/HBV免疫组化检测,记录不同组织每高倍视野(×400)pcDNA3.1/HBV表达阳性细胞数。结果 第1、2组肝、肾组织内偶见pcDNA3.1/HBV表达弱阳性细胞,肺、心、脾、骨骼肌组织未见表达阳性细胞。第3组肝组织内可见少量pcDNA3.1/HBV阳性细胞,每高倍视野表达阳性细胞(26.5±3.9)个。肾、肺、心、脾、骨骼肌组织未见表达。第4组肝组织内pcDNA3.1/HBV表达最强,每高倍视野表达阳性细胞(84.2±4.4)个,为第3组的约3.2倍。肾、肺、心、脾、骨骼肌组织未见表达。结论 静脉注射黏附质粒的微泡声学造影剂同时经体表给予一定强度的超声辐照,能显著增强辐照部位局部组织的基因转染与表达,有望成为一种安全、高效的体内基因定位转染新技术。 |
| 英文摘要: |
| Objective To investigate whether local ultrasound irradiation could enhance the gene delivery to mice livers after intravenous injection of microbubbles in vivo. Methods Twenty-four Kunming mice were randomly divided into four groups, each group with 6 mice. The first group was taken as simple plasmid DNA delivery group: 2 ml of pcDNA 3.1/HBV (20 μg) was quickly injected through the tail vein. The second group was taken as simple microbubbles delivery group: 2 ml of the mixture of microbubbles and plasmid (20 μg) was quickly injected through the tail vein. The third group was taken as simple ultrasound delivery group: after injection of 2 ml plasmid through the tail vein, ultrasound with the frequency of 1 MHz, and the intensity of 0.5 W/cm2 was applied on the livers of mice for 1 min. The last group was taken as ultrasound and microbubbles delivery group: after injection of 2 ml mixture of microbubble and plasmid (20 μg), ultrasound with the same energy was applied on the livers for 1 min. Seven days later, the mice were killed and the livers, kidneys, lungs, hearts, spleen, skeletal muscles were harvested for immunochemical staining of HBcAg. The numbers of positive expression cells in every high power fields were measured. Results There were few positive expression of pcDNA3.1/HBV cells in livers and kidneys in Group 1 and Group 4, while no positive cells in lungs, hearts, spleens, skeletal muscles. In Group 3, there were a few positive cells in the livers and no positive cell in the kidneys, lungs, hearts, spleens, skeletal muscles. The number of positive cells in Group 3 were (26.5±3.9)/HP. The expression of pcDNA3.1/HBV was the highest in Group 4, the number of positive cells was (84.2±4.4)/HP. It was 3.2 folds higher than that in Group 3. There was no expression in kidneys, lungs, hearts, spleens, skeletal muscles in this group. Conclusion Local ultrasound irradiation after intravenous injection of plasmid adhered on the microbubble could highly enhance the local gene transfection and expression. This method may provide a safe, high effective local gene delivery technique in vivo. |
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