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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 investigate the therapeutic effect of small interfering (siRNA) targeting two different sites of rat Lep gene on hepatic fibrosis. Methods Forty male Sprague - Dawley rats were randomly divided into 4 groups: model group (group A) with hepatic fibrosis and subcutaneous injection of 3 ml / kg abdominal wall with 60% CCl4 (exquisite olive oil) twice a week for 6 weeks; The siRNA1 group (group B) was given siRNA1 plasmid 0.2mg / kg twice a week for 6 weeks after the model was prepared. siRNA2 treatment group (group C) 0.2mg / kg twice a week for 6 weeks. The siRNA plasmids were conjugated to Lipofectamine before administration and injected into the body through the tail vein. The normal control group (group D) was injected subcutaneously with purified olive oil instead of CCl4, 2 times, a total of 6 weeks. SiRNA targeting LepmRNA gene at positions 90 and 144 were designed as siRNA1 and siRNA2, respectively, and their expression vectors were constructed. Transfected into CCl4-induced hepatic fibrosis rats by tail vein injection by Lipofectamin method, PCR and immunohistochemistry to detect the expression of Lep and type I and type III collagen in liver tissue and compare the curative effect of different siRNA on experimental liver fibrosis. Results After LepsiRNA transfection, the expression of Lep and type Ⅰ and type Ⅲ collagen decreased, but the effect of group B was better than that of group C. Conclusion siRNA targeting different sites of Lep gene has different therapeutic effect on liver fibrosis.