将含有报告基因LacZ的重组腺病毒载体从大鼠鼻腔转移至脑的实验研究

An adenoviral vector with a report gene Lac Z transferred to brain from nasal cavity of adult rats

目的探讨将携带有报告基因的腺病毒载体由鼻腔转移至脑途径的可行性。方法将携带半乳糖苷酶报告基因(LacZ)的5型重组腺病毒载体(Ad5 CMVLacZ)注入SD大鼠鼻腔黏膜,分别在注射3 d、7 d、14 d、21 d及28 d切取大鼠鼻腔黏膜以及与嗅觉信息传导途径相关的脑组织,进行冰冻切片和β-半乳糖苷酶(β-gal)免疫组织化学反应显色,判断该载体是否转染和表达蛋白质产物。结果β-gal免疫组化染色反应显示,该病毒载体从第3天起即可在鼻腔黏膜与嗅球内的细胞转染和表达,在第7天至第21天病毒载体逐渐向颅内深部的室管膜下区、杏仁核及齿状回细胞转移和表达,持续性表达蛋白质产物达28 d之久。结论经鼻腔黏膜注入腺病毒载体不仅可将外源性基因转移到脑实质内,还可确保外源性基因转染和表达成功。

Objective To explore the feasibility of transferring an adenoviral vector carrying report gene Lac Z(Ad5 CMV Lac Z) to brain from adult rat nasal cavity.Methods Ad5 CMV Lac Z was injected into the submucosa of rat nasal cality.After 3,7,14,21 and 28 days of nasal cavity injection,the rats were sacrificed to transfuse 4% paraformaldehyde through the aorta.β-galactosidase(β-gal) immunohistochemistry was used to detect the transfection and expression of Ad5 CMV Lac Z.Results After 3 days of injection of Ad5 CMV Lac Z into nasal cavity,the nasal cavity mucosa and olfactory bulb began to be transfected and expressed by the vector.As the periods of infection time extended,more cells transfected and expressed by the vector were found in the subventricular zone of lateral ventricle,amygdaloid nuclei,dentate gyrus;the expressions of Lac Z gene in the above-mentioned three brain areas could be kept for a period of 28 days.Conclusion The injection of adenoviral vector from nasal cavity can realize the transferring of the foreign gene to the brain,and make sure the outside gene transfection and expression.It is suggested that nasal cavity-brain foreign gene delivery is a reliable pathway for minimally invasive gene therapy for brain disorders.