期刊论文

中文

中国动脉硬化杂志

1007-3949

2011

19

1

6-12

国家自然科学基金（81000946） 湖南省教育厅基金（09C835）

本校为第一机构

目的 探讨小干扰RNA(siRNA)沉默Toll样受体4(TLR4)基因对慢性温和应激ApoE-/-小鼠腹腔巨噬细胞促炎症细胞因子表达的影响.方法 针对小鼠TLR4基因设计2条siRNA和一条无关序列,再据每一序列合成两条互补并含siRNA正反义链的DNA,退火后与含GFP的载体pRNAT-H1.1/Adeno相连,转染293A细胞,实时荧光PCR筛选有效的一条siRNA,经腺病毒颗粒包装与病毒扩增并感染293A细胞,G418筛选后,挑取单个阳性克隆放大培养,检测病毒滴度.120只雄性ApoE-/-小鼠随机分为三组:(1)慢性温和应激组;(2)慢性温和应激+siRNA组(10 μL/只,尾静脉注射,每5日一次);(3)慢性温和应激+腺病毒空载体组.每组ApoE-/-小鼠40只,实验动物都以...

Aim To examine the effect of Toll-like receptor 4（TLR4） gene silencing by small interfering RNA（siRNA） on the expression of cytokines produced by peritoneal macrophages in chronic mild stress（CMS） ApoE-/-mice. Methods Two siRNA sequences and one negative control sequence were designed targeting the mouse TLR4 gene.Two complementary single-strand DNA were designed and synthesized based on siRNA sequences.The DNA fragments were annealed and ligated to the GFP expression vector pRNAT-H1.1/Adeno.One siRNA with higher interference efficiency t...