反义真核细胞转化生长因子βⅠ型受体与反义基质金属蛋白酶组织抑制因子-1联合作用对大鼠肝纤维化的影响  

Combined effects of antisense T β RI eukaryotic expressing plasmid and antisense TIMP-I eukaryotic expressing plasmid on rat liver fibrosis

作　　者：郑勇[1] 徐丽红[1] 李睿[1] 周婷[1] 孙侃[1] 常向云[1] 陈卫刚[1] 陈莹[1] 

机构地区：[1]新疆石河子大学第一附属医院消化内科,832008

出　　处：《中华肝脏病杂志》2007年第7期493-497,共5页Chinese Journal of Hepatology

摘　　要：目的观察反义转化生长因子βⅠ型受体（TβRI真核表达质粒与反义基质金属蛋白酶组织抑制因子-1（TIMP-1）真核表达质粒联合作用对实验性大鼠肝纤维化的影响。方法构建大鼠反义真核细胞表达质粒，导入大鼠肝纤维化模型体内，通过I型胶原的免疫组织化学以及苦味酸-酸性品红染色观察两种反义质粒联合作用对大鼠肝纤维化的影响，用目标积分吸光度（A）值表示各实验组动物肝组织中蛋白的表达量。结果反义TβRI疗组、反义TβRI反义TIMP-1治疗组、反义TIMP-1治疗组、pcDNA3．1（＋）空质粒对照组、模型对照组、正常对照组TβRI白的A值分别为：（2．11±0．88）×10^5、（1．06±0．57）×10^5、（3．46±1．14）×10^5、（5．66±2．54）×10^5、（5．19±1．22）×10^5和（0．38±0．27）×10^5；TIMP-1蛋白的A值分别为：（1．10±0．22）×10^5、（0．30±0．12）×10^5、（0．65±0．15）×10^5、（2．05±0．36）×10^5、（1．97±0．28）×10^5和（0．10±0．12）×10^5；I型胶原的蛋白表达的A值分别为：（4．37±1．30）×10，、（0．90±0．32）×10^5、（3．40±0．91）×10^5、（6．90±1．61）×10^5、（7．34±1．68）×10^5和（0．41±0．21）×10s。反义TIMP-1治疗组TIMP-1蛋白表达量显著降低（P〈0．05），反义TβRI疗组TβRI白表达量显著降低（P〈0．05），两种反义质粒可有效抑制相应蛋白的表达；反义TIMP-1表达质粒与反义TβRI达质粒均可减少受损肝脏中I型胶原的沉积（P〈0．05），联合应用可进一步减少受损肝脏中I型胶原的沉积（P〈0．01）。在病理形态学方面的观察，反义TIMP-1表达质粒与反义TβRI达质粒均可使受损肝脏的病理形态有一定改善，联合应用可使受损肝脏的病理形态得到进一步的改善。结论反义TIMP-1表达质粒与反义TβRI达质粒对肝纤维化的发展均有一定的干预作用，联合作用可产生更有�Objective To test the hypothesis that the introduction of antisense transforming growth factor beta receptor I （TβRI） plasmid and antisense tissue inhibitor of matrix metalloproteinase （TIMP-1） eukaryotic expressing plasmid into a rat liver fibrosis model may influence the progression of liver fibrosis. Methods Fragments of TβRI cDNA and TIMP-1 cDNA were obtained by reverse transcription polymerase chain reaction （RT-PCR） and then amplified by nest PCR. pcDNA3.1（＋）-antisense TβRI eukaryotic express- ing plasmid was constructed by directional and inverted joins with the purified linear pcDNA3.1（＋） and the purified fragment of TβRI, as well as, pcDNA3.1 （＋）-antisense TIMP-1 eukaryotic expressing plasmid. Therecombinant was identified by restriction endonuclease digestion and DNA sequence analysis. The recombinant plasmids were encapsulated with Lipofectmine 2000, and then they were injected intraperitoneally into the liver fibrosis model rats. The protein expression of type I collagen was evaluated by immunohistochemistry. VG staining of liver slides of the rats was used for histopathological examination. Results Compared with the empty plasmid control group and the disease control group, the deposition of type I collagen decreased in the three antisense treatment groups： antisense TβRI group （4.37 ± 1.30） × 10^5, P 〈 0.05; antisense TIMP-1 group （3.40 ± 0.91）×10^5,P 〈 0.05; antisense TβRI ＋ antisense TIMP-1 group （0.90 ± 0.32） ×10^5,P 〈 0.01; treatment control group （6.90 ±1.61）× 10^5; disease control group （7.34 ± 1.68）×10^5; and the normal control group （0.41 ±0.21）× 10^5]. Significant differences in the pathological grades of fibrosis were found between the normal control group and the other five groups （P 〈 0.05） and also between the disease control group and the three antisense treatment groups （antisense TβRI group P 〈 0.05; antisense TIMP-1 group P 〈 0.05; antisense TβRI ＋ antisense TIMP-1 group P 〈 0.01）,

关 键 词：肝硬化 DNA 反义 受体 表皮生长因子 金属蛋白酶组织抑制因子-1 

分 类 号：R686[医药卫生—骨科学]