人鼠同源Ⅰ型胶原siRNA对TGF-β_1诱导Ⅰ型胶原表达的抑制效应  

Inhibition of human/rat homology collagen type Ⅰ siRNA on the TGF-β_1-induced expression of collagen type Ⅰ

作　　者：周春华[1] 李欣欣[1] 谭余良[2] 赵海丹[1] 于海[1] 

机构地区：[1]中国人民解放军海军总医院肾内科,北京100037 [2]中国科学院生物物理研究所,北京100101

出　　处：《中国现代医学杂志》2008年第17期2496-2499,共4页China Journal of Modern Medicine

基　　金：北京市自然科学基金资助(No:7063094)

摘　　要：目的观察人鼠同源I型胶原siRNA抑制TGF-β1诱导的Ⅰ型胶原高表达的有效性。方法首先构建人鼠同源的Ⅰ型胶原siRNA表达质粒。转染大鼠系膜细胞,用RT-PCR方法观察对Ⅰ型胶原的直接抑制作用;经不同浓度(1ng、5ng、10ng)的TGF-β1刺激后,再转染Ⅰ型胶原siRNA质粒,RT-PCR方法观察其抑制效应。结果1～4μg的Ⅰ型胶原siRNA质粒均能使COL1A1基因条带表达下调,并随siRNA质粒浓度增加,抑制作用逐渐增强。大鼠系膜细胞经TGF-β1刺激,I型胶原基因表达明显增强,并具剂量依赖性。再转染Ⅰ型胶原siRNA质粒后,TGF-β1+Ⅰ型胶原siRNA质粒各组与相应浓度的单纯TGF-β1各组比较COL1A1基因条带均减弱,显示出抑制效应。结论本实验构建的人鼠同源Ⅰ型胶原siRNA质粒,在大鼠系膜细胞内不仅能够直接抑制Ⅰ型胶原的基因表达,而且对TGF-β1诱导的I型胶原表达增加同样具有较好的抑制作用。本实验结果可望为肾脏纤维化的防治提供思路。[Objective] To observe the inhibition action of human/rat homology collagen type Ⅰ siRNA on human TGF-β1-induced expression of collagen type Ⅰ. [Methods] Construction of vector of human/rat homology siRNA for collagen type Ⅰ. （1）We transfected the established plasmid into the rat mesangial cell and observed its direct re- pression by using RT-PCR. （2） After Stimulated by TGF-β1 with different doses（1 ng, 5 ng, 10 ng）, collagen type I siRNA plasmid with 3 μg transfected into the rat mesangial cell and observed its inhibition action by using RT- PCR. [Results] We can see that collagen type Ⅰ siRNA plasmid with 1-4 μg down-regulate collagen type Ⅰ expres- sion with dosage dependency. By using TGF-β1 induced collagen type I exerted an increased gene expression and showed a more significant effect when the TGF-β1 dosage was increased. After transfection of collagen type Ⅰ siRNA plasmid, compared with corresponding doses of TGF-β1 groups, COL1A1 gene strap of TGF-β1 collagen type Ⅰ siR- NA plasmid groups showed decreased brightness, demonstrated its inhibition action. [Conclusion] Human/rat ho- mology collagen type Ⅰ siRNA plasmid not only direct repress collagen type Ⅰ gene expression in rat mesangial cells, but also have the same inhibition action of TGF-β1-induced overexpression of collagen type Ⅰ. This experimental result would provide a valuable path to prevent and cure kidney fibrosis.

关 键 词：SIRNA I型胶原 TGF 

分 类 号：R-332[医药卫生]