摘要
目的研究针对大鼠瘦素(Lep)基因两个不同位点的小干扰(siRNA)对肝纤维化的治疗效果。方法 40只雄性SD大鼠随机均分为4组:肝纤维化模型组(A组),给予60%CCl4(精致橄榄油配制)3ml/kg腹壁皮下注射,每周2次,共6周;siRNA1治疗组(B组),在模型制备2周后同时给予siRNA1质粒0.2mg/kg,每周2次,共6周;siRNA2治疗组(C组),在模型制备2周后同时给予siRNA2质粒0.2mg/kg,每周2次,共6周,siRNA质粒给予前与Lipofectamin偶联后通过尾静脉注射导入体内;正常对照组(D组),用精制橄榄油溶液代替CCl4皮下注射,每周2次,共6周。设计针对LepmRNA基因90和144位点的siRNA,分别记为siRNA1、siRNA2,并构建其表达载体;用Lipofectamin方法通过尾静脉注射转染入CCl4诱导的肝纤维化大鼠体内;然后分别采用RT-PCR、免疫组化方法检测Lep与Ⅰ、Ⅲ型胶原在肝脏组织中的表达,比较不同siRNA对实验性肝纤维化的疗效。结果转染LepsiRNA后Lep和Ⅰ、Ⅲ型胶原蛋白的表达均减少,但B组疗效优于C组。结论针对Lep基因不同位点的siRNA对肝纤维化治疗效果不同。
Objective To study and compare the effects of two small interfering RNAs（siRNAs） targeting against different sites of leptin gene on liver fibrosis between.Methods The model of hepatic fibrosis in rats was made by injection of carbon tetrachloride（CCl4）.Forty male SD rats were randomly divided into two treating groups of hepatic fibrosis model（group siRNA1 and group siRNA2）,model control（group A）,and normal control（group D） with 10 rats each.The different small interfering RNAs,targeting leptin gene 90 and 144 were designed with computer according to analysis of the secondary structure of leptin gene.Leptin and Ⅰ,Ⅲ collagen were detected by immunohistochemistry and RT-PCR,respectively.Results The levels of leptin andⅠ,Ⅲ collagen were significantly decreased in gorups of siRNA1 and siRNA2 ,which was more in group siRNA2 than that in group siRNA1.Conclusion The siRNA targeting against different sites of leptin gene has different therapeutic effects on hepatic fibrosis.
出处
《江苏医药》
CAS
CSCD
北大核心
2010年第18期2192-2195,共4页
Jiangsu Medical Journal
基金
南通市社会发展基金项目(S2007039)
