【作者】 唐海英；

【导师】 吴泰华；

【作者基本信息】 大连医科大学 ， 内科学， 2006， 硕士

【摘要】 背景与目的:肺纤维化是由于各种急性和慢性肺损伤后引起细胞外基质(Extracellular matrix,ECM)成分组成及量的改变、ECM在肺中病理性沉积,造成肺功能异常的重要因素。肺纤维化的过程是一个非常复杂的过程。虽然引起肺纤维化的病因或原发病因有很大差异,但是肺纤维化过程中ECM代谢异常是类似的。如果肺内ECM的合成增加,或降解减少,或二者都有,都能引起ECM积聚,进而导致肺纤维化形成。然而,现仍无有效的治疗方法。近年来的研究表明ECM降解减弱是引起ECM积聚的重要因素,认为肺纤维化过程中基质金属蛋白酶组织抑制因子-1 (Tissue inhibitors of metalloproternase-1,TIMP-1)表达增高,抑制了基质金属蛋白酶(Matrix metalloproteinases,MMPs)的活性,使ECM降解减少,促进肺纤维化的发生发展,因此抑制TIMP-1的表达是阻止肺纤维化发生、发展的重要环节。本研究以博来霉素(Bleomycin, BLM)诱导的肺纤维化大鼠为模型,观察肺纤维化过程中基质金属蛋白酶-2(Matrix metalloproteinase-2,MMP-2)及TIMP-1基因及蛋白表达的变化。在实验模型观察的上述变化的基础上,应用反义核酸技术将正、反义TIMP-1cDNA逆转录病毒载体转染到不同阶段肺纤维化大鼠模型肺内,观察内源性TIMP-1的表达变化,了解外源基因治疗性质粒的疗效,探讨基因治疗肺纤维化的可行性及有效性,为今后临床肺纤维化的基因治疗提供理论依据。方法: 1.实验动物:6周龄SD大鼠,雌性,体重180-220g 2.动物模型制备:大鼠经乙醚麻醉后,颈前暴露气管,一次性气管内注入0.4% BLM生理盐水注射液(5mg/kg),均由腹主动脉取血处死更多还原

【Abstract】 Objective: Pulmonary fibrosis is a progressive pathological process, initiated by pulmonary parenchymal damage and perpetuated by inflammatory reaction. It is predominantly characterized by excessive accumulation of extracellular matrix (ECM) components in the lung. The process of pulmonary fibrosis formation is very complicated. The Reason of producing pulmonary fibrosis is different, but it is analogical about ECM metabolism abnormality. Excessive ECM deposition resulting from ECM degradation overwhelmed or ECM synthesis increased leads to pulmonary fibrosis. However, no effective treatment of pulmonary fibrosis is available clinically at present.Previous studies show that decreased degradation of ECM plays an important role in the process of excessive accumulation of ECM in recent years. Internal and foreign scholars all think that increasing the expression of tissue inhibitors of metalloproternases (TIMPs), in particular TIMP-1, suppresses the activity of matrix metalloproternases (MMPs), which contributes to the formation of pulmonary fibrosis by decreasing the degradation of ECM. Thus, it is also important to prevent fibrogenesis by suppressing the expression of TIMP-1. To study the roles of MMP-2 and TIMP-1 in pulmonary fibrosis, the protein and gene expressions of MMP-2 and TIMP-1 in rat pulmonary fibrosis models induced by bleomycin (BLM) were observed and recombinant retroviral vectors containing sense and antisense hTIMP-1 respectively were transfected into fibrotic lungs of different stages in vivo to investigate the changes of the endogenous TIMP-1 expression. The aim of this study is to observe the effectiveness of更多还原