摘要
目的 :研究小鼠博来霉素在肺纤维化模型中肺泡巨噬细胞(AM)分泌的基质金属蛋白酶 9(MMP 9)和基质金属蛋白酶抑制剂 1(TIMP 1)的表达随着时间的变化 ,观察肺纤维化中MMP 9和TIMP 1的表达是否存在失衡。方法 :以博来霉素诱导建立小鼠肺纤维化模型 ,采用HE染色观察肺部胶原沉积状况。选用抗CD6 8mAb ,采用微小免疫磁珠法分离纯化肺泡灌洗液中的AM ,用Hoechst 332 5 8染色AM ,在下光镜高倍视野计数法评估AM的纯度和活性 ,用EIA法检测AM上清液中MMP 9和TIMP 1的表达。结果 :肺组织切片HE染色显示小鼠肺部胶原沉积在 1、3、7、14、2 8d呈逐渐加重趋势。粗分离的AM纯度为 (82 .5± 2 .5 ) %。采用微小免疫磁珠法分离肺泡灌洗液中的AM纯度可达到 (99.3± 0 .7) % (P <0 .0 5 )。纯化后的细胞存活率为 (92 .5± 1.8) % ,与纯化前的 (92 .7± 2 .0 ) % ,相差不大 (P >0 .0 5 )。随着小鼠肺间质纤维化的进展 ,MMP 9的分泌量逐渐减少 ,而TIMP 1的表达量逐渐增加。结论 :在小鼠肺纤维化中AM分泌的MMP 9和TIMP 1之间存在失衡 。
AIM: To evaluate the balance between matrix metalloprotease-9 (MMP-9) and its inhibitor, tissue inhibitor of metalloproteinases-1 (TIMP-1) in bleomycin-induced pulmonary fibrosis in mice. METHODS: Pulmonary fibrosis model was induced by bleomycin in mice. The histological images of lungs were studied by HE staining. Alveolar macrophages(AMs) were separated by anti-CD68 mAbs using immunomagnetic beads(d≤50 nm). The purity of AMs was determined under microscope. Cell viability was evaluated by staining with Hoechst 33258 dye. The secreted MMP-9 and TIMP-1 were assayed in macrophage supernatants, (using) quantitative enzyme-linked immunosorbent assays (EIA). RESULTS: Collagen deposition was gradually (increased) in the lung tissue of mice on days 1, 3, 7, 14 and 28 after bleomycin treatment. The purity of AMs could be increased from (82.5±2.5)% to (99.3±0.7)% by mini-magnetic cell sorting system (mini-MACS, P<0.05), and the rate of living cells post-MACS was (92.5±1.8)% (pre-MACS (92.7±2.0)%, P>0.05). The concentration of MMP-9 secreted by AMs was gradually decreased on days 1, 3, 7, 14 and 28, and levels of TIMP-1 gradually increased. CONCLUSION: There was an imbalance between Macrophage-derived MMP-9 and TIMP-1 in bleomycin-induced pulmonary fibrosis in mice.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第6期723-726,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
西安市农医社会发展计划项目资助 (No .SF2 0 0 336)
