TIMP-1siRNA对肝纤维化大鼠TIMP-1及胶原表达的影响  

Effect of TIMP-1 siRNA on the expression of TIMP-1 and collagen in rats with liver fibrosis

作　　者：张燕[1] 郎清[1] 石小枫[1] 刘杞[1] 孙航[1] 

机构地区：[1]重庆医科大学附属第二医院感染科教育部感染性疾病分子生物学重点实验室,重庆400010

出　　处：《重庆医科大学学报》2016年第4期364-368,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金面上资助项目(编号:81270503)

摘　　要：目的:探讨基质金属蛋白酶抑制因子-1(tissue inhibitor of metalloproteinases-1,TIMP-1)的siRNA经体内转染后对CCl4诱导的肝纤维化大鼠TIMP-1及胶原表达的影响。方法:将32只雄性SD(Sprague Dawley)大鼠随机分为4组:正常组、模型组、TIMP-1 siRNA处理组和TIMP-1 siRNA阴性对照组。CCl4皮下注射诱导大鼠肝纤维化模型,共12周。HE染色验证肝纤维化程度,免疫组织化学染色检测TIMP-1和Ⅰ、Ⅲ型胶原的表达;Western blot检测TIMP-1的蛋白表达,qRT-PCR法检测TIMP-1和Ⅰ、Ⅲ型胶原的mRNA表达。结果:病理组织学显示相较模型组、阴性对照组,肝组织肝纤维化程度在TIMP-1 siRNA处理组中明显减轻。TIMP-1和Ⅰ、Ⅲ型胶原的表达量也明显减少(P=0.000)。结论:TIMP-1 siRNA能抑制或阻断TIMP-1表达,进而促进以Ⅰ、Ⅲ型胶原为主的细胞外基质的降解,肝纤维化程度得到改善,对CCl_4诱导的大鼠肝纤维化有一定的防治效果。Objective：To investigate the expression of tissue inhibitor of metalloproteinases-1（TIMP-1）and type Ⅰ,Ⅲ collagen in carbon tetrachloride（CCl4）- induced liver fibrosis rats treated with small interference RNA（siRNA）targeting TIMP-1. Methods：A total of 32 male SD（Sprague Dawley）rats were randomly and averagely divided into 4 groups：normal group,model group,TIMP-1siRNA treatment group（0.25 mg/kg）,TIMP-1 siRNA negative control group（0.25 mg/kg）. CCl4 was used for 12 weeks to induce liver fibrosis. All rats were then sacrificed to collect liver tissue samples. A portion of the liver samples was soaked in formalin for testing the degree of liver fibrosis by hematoxylin-eosin staining. The expression of TIMP-1 and type Ⅰ,Ⅲ collagen was detected by immuno histochemical staining. TIMP-1 protein expression was detected by Western blot and TIMP-1 mRNA expression and type Ⅰ,Ⅲ collagen was measured by quantitative real-time polymerase chain reaction. Results：Compared with those in model group and negative control group, the tissue of liver in TIMP-1 siRNA treatment group has normal structure, with a small number of inflammatory cell infiltrating,and there is no obvious proliferation of collagen fibers. Immunohistochemistry showed the protein expressions of TIMP-1 and type Ⅰ,Ⅲcollagen were decreased significantly. Western blot showed the protein expression of TIMP-1 was decreased significantly. Real-time PCR showed that the mRNA expression of TIMP-1,type Ⅰcollagen and type Ⅲ collagen were decreased significantly. Conclusion：TIMP-1 siRNA can inhibit the expression of TIMP-1,and can effectively prevent the occurrence and development of liver fibrosis induced by CCl_4 in rats. The mechanism is the down-regulation of TIMP-1 expression can promote extracellular matrix degradation.

关 键 词：基质金属蛋白酶抑制因子1 小干扰RNA 肝纤维化 基因治疗 

分 类 号：R575.2[医药卫生—消化系统]