门静脉及股静脉注射对导入基因在肝脏表达影响的比较  

A comparative study on the expression of hepatic stimulator substance in liver via different gene delivery routes

作　　者：吴媛[1,2] 俞豪 董凌月[1,2] 王欣 李紫薇[1,2] 安威 Wu Yuan;Yu Hao;Dong Lingyue;Wang Xin;Li Ziwei;An Wei(Department of Cell Biology, Capital Medical University,Beijing 100069, China;Municipal Laboratory for Liver Protection and Regulation of Regeneration,Beijing 100069, China)

机构地区：[1]首都医科大学基础医学院细胞生物学系 [2]肝脏保护与再生调节北京市重点实验室,北京100069

出　　处：《中国组织化学与细胞化学杂志》2017年第6期559-565,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助项目(31371169);首都医科大学科研基金-校基金(技术类)(2016JS18)

摘　　要：目的研究外源基因的门静脉途径和股静脉途径注射对导入基因在肝脏内表达和分布的影响。方法分别采用门静脉和股静脉的注射方法将包装好的连有目的基因的腺病毒载体导入小鼠体内。在注射后不同时间处死小鼠,分离肝脏,通过血清生化指标和HE染色检测评价两种方法对肝脏的损伤,利用免疫组织化学、Real-time PCR和Western blot等方法测定目的基因和载体标签蛋白在肝脏内的表达及分布情况。结果注射3d后,血清生化指标检测结果显示门静脉注射组丙氨酸转氨酶(alanine aminotransferase,ALT)和天冬氨酸转氨酶(aspartate aminotransferase,AST)水平明显高于股静脉注射组;HE染色结果同样表明门静脉注射组肝组织内出现炎性细胞浸润,较股静脉注射组更明显;Real-time PCR、Western blot及免疫组织化学检测均显示门静脉注射后,目的基因在肝脏内的表达明显早于股静脉注射组,但至注射后7d时两组目的基因的表达水平已无明显差异。结论与门静脉注射相比,股静脉注射外源基因对肝脏基本没有损伤,目的基因的表达虽然较晚,但也能达到门静脉注射水平。Objective To evaluate the influences of different gene delivery routes on exogenous gene expression in liver.Methods Adenoviral vectors carrying genes of hepatic stimulator substance(HSS)-Flag fusion protein were injected into mice through portal or femoral vein.The mice were sacrificed3to7days after injection and their livers were isolated.Liver injury was evaluated by HE staining as well as the serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST).The expression levels and patterns of HSS-Flag were determined by immunohistochemistry,real-time PCR and western blotting.Results Three days after viral vector injection,compared to femoral vein route,there were marked increases of serum ALT and AST activities in mice administered through portal vein.HE staining revealed mice in portal vein injection group had more inflammatory cell infiltration in liver tissues.Results from immunohistochemistry,real-time PCR and Western blotting indicated that the expression levels of HSS-Flag fusion protein were higher in mice injected through portal vein compared to those through femoral vein3and5days after injection.However,this difference diminished7days after injection.Conclusion Compared to portal vein route,viral vector injection through femoral vein caused less liver damage,slower but similar gene expression level at peak.

关 键 词：基因治疗 门静脉注射 股静脉注射 

分 类 号：Q503[生物学—生物化学]