携带EGFP基因的腺病毒在大鼠脑和肺的表达
Expression of the Recombinant Adenovirus Carrying Enhanced Green Fluorescent Protein(EGFP)in Brain and Lung Tissues of Rats
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摘要: [摘要]目的 探讨携带增强型绿色荧光蛋白(EGFP)基因重组腺病毒经大鼠鼻腔给药后在大鼠脑部和肺部的表达,为腺病毒经鼻腔给药的靶向性研究提供依据;并对比腺病毒液加入粘性纤维蛋白原液,是否可有利于腺病毒在大鼠脑部和肺部的表达.方法 大鼠12只,随机分为3组,每组4只,分别鼻腔滴入EGFP重组腺病毒液、EGFP重组腺病毒纤维蛋白原液和对照生理盐水.腺病毒组每次每只鼻孔滴入20 μL病毒液;腺病毒纤维蛋白原液组每日每只鼻孔注入40 μL病毒蛋白胶溶液;生理盐水组每日每只鼻孔注入20 μL生理盐水,每日1 次, 连续滴鼻7 d,第9天对大鼠进行心脏灌注, 取脑组织及肺组织做冰冻切片,共聚焦荧光显微镜下观察冰冻切片荧光表达.结果 在2组给药大鼠的肺切片中均观察到了绿色荧光的表达,而在大鼠脑部各部位的切片中未看到明显的绿色荧光表达.结论 腺病毒通过鼻腔给药可以到达大鼠肺部,可作为腺病毒携带基因治疗肺部靶向给药的载体.Abstract: [Abstract]Objective To investigate the expression of the recombinant adenovirus carrying the enhanced green fluorescent protein(EGFP)in rats' brain and lung by nasal administration,so as to provide a basis for targeting tvansport of adenovirus by intranasal administration.Methods 12 rats were randomly divided into 3 groups(n = 4),rats in 3 groups were respectively given instillation of EGFP recombinant adenovirus fluid, EGFP recombinant adenovirus added by viscous fiber protein solution and control group instillation of saline by nasal administration. Rats in the adenovirus fluid group were given instillation of 20 μL adenovirus fluid from each nostril.Rats in the adenovirus added by viscous fiber protein liquid group were given instillation of 40 μL adenovirus added by viscous fiber protein liquid each nostril. Rats in the control group were given instillation of 20 μL saline.The administration was once a day and lasted for 7 days.Rats were given heart perfusion on the 9th day,and lung and brain tissues were collected for frozen section,and the fluorescence signals in frozen sections were observed by confocal fluorescence microscopy.Results We observed the expression of green fluorescent protein in the lung tissues of rats in the two treatment groups,but no green fluorescent expression in brain tissues. Conclusion Adenovirus can target rats' lungs by intranasal administration,so it can be used for the carrier of pulmonary targeted drug delivery system.
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