摘要
目的观察反义TIMP-1真核表达质粒对实验性肝纤维化的影响。方法运用重组DNA技术及反义技术,构建反义TIMP-1真核细胞表达质粒,经脂质体Lipofectmine2000包埋。采用腹腔注射将其导入到复合因素复制的肝纤维化大鼠模型体内,通过半定量RT-PCR,病理形态学观察及免疫组化等方法,观察反义TIMP-1表达质粒对大鼠肝纤维化的影响。结果反义TIMP-1质粒组与肝纤维化组、空质粒组相比TIMP-1、MMP-13 mRNA及蛋白表达明显减少(P<0.05),空质粒组与肝纤维化组无显著性差异(P>0.05);正常对照组与各实验组之间的病理学分级有显著性差异(P<0.05);反义TIMP-1质粒组病理学分级较空质粒组有明显改善(P<0.05);空质粒组与肝纤维化组无显著性差异(P>0.05)。结论反义TIMP-1/pcDNA3.1(+)真核细胞表达质粒使TIMP-1、MMP-13 mRNA及蛋白的表达受到抑制,反义TIMP-1表达质粒对肝纤维化有较好的改善作用,并为以后肝纤维化的基因治疗奠定基础。
Objective To observe the effect of antisense tissue inhibitor of metalloproteinase-1 （TIMP-1） expressing plasmid on rat liver fibrosis. Methods RT-Nest-PCR, gene recombinant techniques and antisense techniques were used to construct the rat antisense TIMP-1 recombinant plasmid which could be expressed in eukaryotic cells. It was encapsulate by Lipofectmine 2000. Antisense TIMP-1 recombinant plasmid was injected into rats abdominal cavity of composite factors duplicated liver fibrosis model. Expression of exogenous transfected plasmid was assessed by semi-RTPCR, pathomorphology study and immunohistochemistry. Results Compared with the treatment group and treatment control group,disease control group expression of TIMP-1 mRNA, MMP-13 mRNA and protein all decreased （P 〈 0.05）. No difference was found in expression of all the three mRNA and protein between treatment control group and disease control group. A significant difference was found in the pathologic grade between normal control group and the other three groups. A significant difference was found between treatment group and treatment control group. Same difference was found between treatment group and disease control group（P 〈 0.05）. No difference was found in pathologic grade between treatment control group and disease control group. Conclusions Antisense TIMP-1 eukaryotic expressing plasmid could inhibit the expression of TIMP-1 mRNA, MMP-13 mRNA and protein in rat liver fibrosis. The antisense TIMP-1 recombinant plasmid has certain extern reverse effect on liver fibrosis and it is maked as a possible candidate for use in future gene therapy.
出处
《胃肠病学和肝病学杂志》
CAS
2008年第4期325-328,共4页
Chinese Journal of Gastroenterology and Hepatology
基金
新疆生产建设兵团重大科技攻关项目(04GG22)
