比较不同方法肝脏靶向性导入抗TGF-β_1 siRNA对肝细胞TGF-β/Smad信号传导通路的影响  

Effect of transfecting anti-TGF-β_1 siRNA on TGF-β/Smad signal transmission pathway in the liver cells by different hepatic targeting methods

作　　者：张涛[1] 黄顺玲[2] 孙克伟[1] 谭德明[2] 

机构地区：[1]湖南中医药大学第一附属医院传染科,湖南长沙410007 [2]中南大学湘雅医学院感染科

出　　处：《胃肠病学和肝病学杂志》2011年第2期132-137,共6页Chinese Journal of Gastroenterology and Hepatology

基　　金：湖南省科技计划项目(2007FJ4176);湖南省卫生厅中医药科研基金(2006103)

摘　　要：目的比较抗TGF-β1 siRNA靶向性进入肝细胞的4种转染方式在抗肝损伤与纤维化及对TGF-β/Smad信号传导通路影响中的效果差异,并探讨各自的优劣。方法已启动肝组织中的TGF-β/Smad信号传导通路的50只模型鼠,随机分为模型对照组、尾静脉直接注射组、脂质体包裹组、流体动力学法组和肝动脉插管组,每组10只;将构建的TGF-β1 siRNA质粒分别通过尾静脉直接注射、脂质体包裹、流体动力学法、肝动脉插管等方式导入肝脏靶细胞;分别检测血清指标ALT、AST、HA;取肝组织,HE染色病理检测;RT-PCR技术检测Ⅰ、Ⅲ型胶原、TGF-β1和Smad3 mRNA表达;免疫组化技术检测Ⅰ、Ⅲ型胶原、TGF-β和Smad3蛋白表达。结果抗TGF-β1siRNA质粒经过以上4种导入方法转染入靶细胞,均能有效地阻断大鼠肝组织中的TGF-β/Smad信号传导通路的传导(P<0.05);4种导入方法比较,肝动脉插管组与脂质体包裹组效果相当(P>0.05),优于尾静脉直接注射组及流体动力学法组(P<0.05),尾静脉直接注射组效果最差(P<0.05)。结论抗TGF-β1siRNA质粒经肝动脉插管途径转染,是一种安全、高效的外源基因肝脏靶向导入转染方法。Objective To compare the difference of effect on the anti-hepatic injury and fibrosis and the influence of TGF-β/Smad signal transmission pathway with four transfection methods by anti-TGF-β1 siRNA entered target cells.Methods The 50 rats priming TGF-β/Smad signal transmission pathway in the liver tissues were randomly divided into 5 groups： the model control group（group M,n=10）,the vena caudalis straight injection group（group V,n=10）,the liposome package group（group L,n=10）,the hydrodynamics group（group H,n=10） and hepatic artery catheterization group（group A,n=10）.TGF-β1 siRNA plasmids were introduced into target cells in the liver by different methods. Serum ALT,AST,HA were detected to evaluate the effect of four transfection methods;liver tissue was detected by HE staining;expression of type Ⅰ and Ⅲ collagen,TGF-β1 and Smad3 mRNA were detected by RT-PCR;expression of type Ⅰ and Ⅲ collagen,TGF-β1 and Smad3 protein were detected by immunohistochemistry technology.Results The anti-TGF-β1 siRNA plasmids transfected into target cells by each method effectively blocked the TGF-β/Smad signal transmission pathway in the rats＇ hepatic tissues（P0.05）.The effect in group L and group A was significantly better than that in the group V and group H（P0.05）,there was no significant difference between group A and group L（P0.05）.The effect in the group V was the worst （P0.05）.Conclusion The anti-TGF-β1 siRNA plasmid transfecting by hepatic artery catheterization is a safe and high-performance method for hepatic targeting transfection of foreign gene.

关 键 词：肝脏靶向性给药 小干扰RNA TGF-β/Smad信号传导通路 基因转染 

分 类 号：R575.2[医药卫生—消化系统]