瘦素特异性小干扰RNA对实验性肝纤维化的影响  

Effects of decreased leptin expression on liver fibrosis

作　　者：冯海娟[1] 朱净[1] 潘亮[1] 陆静贤[1] 肖明兵[1] 黄华[1] 倪润洲[1] 陆翠华[1] 

机构地区：[1]南通大学附属医学院消化内科,226000江苏南通

出　　处：《中华肝脏病杂志》2010年第5期342-345,共4页Chinese Journal of Hepatology

摘　　要：目的 研究针对大鼠瘦素基因的小干扰（si）RNA对肝纤维化的治疗效果.方法 将40只雄性SD大鼠平均分为4组,正常对照组皮下注射橄榄油溶液3 ml/kg,2次/周,共6周;肝纤维化模型组皮下注射60% CCl4 3 ml/kg,2次/周,共6周;siRNA治疗组和阴性对照组均于皮下注射60% CCl4 3 ml/kg,2周后尾静脉分别注射siRNA或阴性对照质粒0.2 mg/kg.每周测体质量1次,根据体质量调整药物用量.设计针对瘦素mRNA的siRNA,并构建其表达载体;用脂质体方法通过尾静脉注射转染入CCl4诱导的肝纤维化大鼠体内,然后分别采用逆转录-聚合酶链反应、免疫组织化学方法检测瘦素与Ⅰ、Ⅲ型胶原在肝脏组织中的表达.免疫组织化学资料采用等级资料秩和检验.PCR结果采用单因素方差分析进行统计学处理,P＜0.05时用SNK法进行多重比较.结果 瘦素mRNA在正常对照组组、肝纤维化模型组、siRNA治疗组和阴性对照组的相对表达量分别为0.75±0.03、2.34±0.14、0.90±0.06、2.31±0.13,各组比较,F=797.64,P=0.000,差异有统计学意义;Ⅰ型胶原mRNA在各组的相对表达量分别为1.10±0.11、2.75±0.13、1.27±0.08、2.77±0.12,各组比较,F=695.71,P=0.000,差异有统计学意义;Ⅲ型胶原mRNA在各组的相对表达量分别为0.62±0.07、1.52±0.09、0.85±0.08、1.51±0.07,各组比较,F=371.57,P=0.000,差异有统计学意义.结论 针对瘦素基因的siRNA对肝纤维化有明显的治疗作用.Objective To study the effects of decreased leptin expression on liver fibrosis. Methods The small interfering RNA, targeting leptin gene, was designed according to the secondary structure of leptin gene. The recombinant plasmids were encapsulated with lipofectamin and then injected into carbon tetrachloride （CCl4） induced rat liver fibrosis models. Leptin and Ⅰ, Ⅲ collage were detected by immunohistochemistry and reverse transcription polymerase chain reaction （RT- PCR）. Results The mRNA and protein levels of leptin in the fibrotic liver transfected with leptin shRNA were significantly decreased compared with those in controls （P 〈 0.01）.The depositions of type Ⅰ and type Ⅲ collagens were also decreased （P 〈 0.01）. Conclusion Decreased leptin expression prevents liver fibrosis.

关 键 词：肝硬化 瘦素 大鼠 模型 动物 

分 类 号：R285.5[医药卫生—中药学]