半乳糖配体介导大鼠白介素10基因在大鼠肝脏内的靶向表达  

Galactose Ligand-mediated Targeting Expression of Rat Interleukin10 Gene in Rat Liver

作　　者：黄月红[1] 陈运新[1] 郑伟达[1] 张莉娟[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福州市350001

出　　处：《医学分子生物学杂志》2011年第1期32-35,共4页Journal of Medical Molecular Biology

基　　金：福建省自然科学基金（No.2010J05062）;福建省卫生厅青年科研项目（No.2010-1-10）

摘　　要：目的 探讨大鼠白介素10（rIL-10）基因是否可通过半乳糖配体介导的脂质体转染法在大鼠肝脏内靶向表达。方法将已构建好的rIL-10基因真核表达质粒与半乳糖配体转染试剂按jetPEI．Gal／DNA（N／P=10）比例混合，通过尾静脉注射转移至大鼠体内。RT—PCR法和ELISA法检测rIL-10基因转移至体内0h、24h、7d和16d后大鼠肝、肾、脾和肺组织及血清中rIL-10的表达情况。结果rIL-10基因转移前大鼠肝、肾、脾和肺组织末扩增出明显rIL-10mRNA表达，转移7d后rIL-10表达主要分布在肝组织。肝组织中rIL-10mRNA表达在基因转移24h和7d时显著升高。血清中的rIL-10浓度在转移后24h和7d浓度分别为（107．92±12．26）pg／ml和（33．2±13．15）pg／ml。结论rIL-10基因通过半乳糖配体介导的脂质体转染法可有效的转移至大鼠体内，并可在肝脏靶向表达一周左右时间。Objective To investigate targeting expression of rat interleukin 10 （rIL-10） gene in the rat liver by use of galactose ligand-mediated liposome transfection. Methods The established rIL-10 eukaryotic expression plasmid was mixed with galactose ligand-mediated liposome transfection reagent at a PEI nitrogen-to-DNA phosphate charge ratio 10 （N/P = 10） . The mixture was injected via tail vein into rats. At the time points 0 h, 24 h, 7 d and 16 d after gene transfer, the expression of rIL-10 gene in the liver, kidney, spleen and lung tissue and the serum was detected by RT- PCR and ELISA analysis respectively. Results After 24 h and 7 d injection, the levels of rIL- 10mRNA were increased significantly in the liver. The level of rIL-10mRN was detected in the serum at （ 107.92 ±12.26） pg/ml after 24 h and maintained at （ 33.2 ±13.15 ） pg/ml after 7 d. rIL- 10mRNA expression was not significantly increased in the kidney, spleen and lung tissue. Conclusion rIL-10 gene can be effectively transferred to rat liver by use of galactose ligandmediated liposome transfection. And targeting expression of rIL-10 gene in the liver can be sustained for one week.

关 键 词：大鼠白介素10 基因转移 配体介导脂质体转染 靶向表达 

分 类 号：Q786[生物学—分子生物学]