摘要
[目的]观察反义金属蛋白酶组织抑制因子-1(TIMP-1)基因转染对博来霉素(BLM)诱导的大鼠肺纤维化肺组织中TIMP-1和Ⅰ型胶原的影响。[方法]SD大鼠随机分为T1组(正义TIMP-1转染组)、T2组(反义TIMP-1转染组)、T3组(空载体转染组)以及C组(正常对照组)、C1组(肺纤维化组)。C组不给予任何干预措施;T1、T2、T3组在给予BLM后第1、3、7、14、28、60、90天分别将含正、反义人TIMP-1cDNA逆转录病毒载体、空载体导入大鼠肺内,转染后28 d处死大鼠;C1组仅给予BLM,时间与转染组相同。HE染色、Masson染色法观察肺组织病理形态学变化,RT-PCR和免疫组织化学检测TIMP-1和Ⅰ型胶原的表达。[结果]给予BLM后第1、3天T2组与同一时间点的C1组及T3组比较纤维化程度减轻,TIMP-1和Ⅰ型胶原基因和蛋白的表达下降(P<0.05);而T1组纤维化程度加重,TIMP-1和Ⅰ型胶原基因和蛋白的表达增高。给予BLM后第7、14、28、60、90天T1和T2组与同一时间点的C1组及T3组比较均无明显不同。[结论]给予BLM后第1、3天进行反义TIMP-1基因转染可以在一定程度上抑制肺纤维化的发生和发展。
[Objective] To observe the effect of antisense tissue inhibitors of matrixmetalloproteinase-1（TIMP-1） cDNA on expression of TIMP-1,type Ⅰ collagen in tissue of pulmonary fibrosis induced by bleomycin（BLM）.[Methods] Sprague Dawley（SD） rats were randomly divided into 5 groups： Group T1（sense TIMP-1 transfection group）,Group T2（antisense TIMP-1 transfection group）,Group T3（empty vector transfection group）,Group C（control gruop） and Group C1（pulmonary fibrosis group）.In GroupT1,T2 and T3,the rats were adminstrated with sense TIMP-1 cDNA,antisense TIMP-1cDNA and empty vector after intratracheal injection of BLM on 1st,3rd,7th,14th,28th,60th and 90th day, then the rats were sacrificed on day 28th days.In Group C1,the rats were adminstrated with BLM at the same timepoints.In Group C,nothing intervening measure was given.HE and Masson staining methods were used to observe the lung pathological changes and the expression of TIMP-1 and type Ⅰ collagen were detected by RT-PCR and immunohistochemistry.[Results] The degree of pulmonary fibrosis of Group T2 were extenuated on 1st,3rd day following intratracheal injection of BLM,but aggravated of Group T1 compared to that of Group T3 and C1.The lung tissue TIMP-1 and type Ⅰ collagen expressions decreased significantly,but increased in Group T1 compared to Group T3 and C1（P〈0.05）.No significant differences were observed in the degree of pulmonary fibrosis and TIMP-1,type Ⅰ collagen expression levels on 7th,14th,28th,60th and 90th day among the T1,T2,T3 and C1 groups.[Conclusion] Transfection with antisense-TIMP-1 on the first and third day can suppress development of pulmonary fibrosis induced by BLM to some extent.
出处
《大连医科大学学报》
CAS
2012年第6期525-531,共7页
Journal of Dalian Medical University
基金
国家自然科学基金项目(30370620)
关键词
肺纤维化
基因转染
金属蛋白酶组织抑制因子-1
Ⅰ型胶原
pulmonary fibrosis
gene transfection
tissue inhibitors of matrixmetalloproteinase-1
type Ⅰ collagen
