小鼠肺纤维化模型中基质金属蛋白酶-9及其抑制剂表达水平的变化

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 ,表明基质降解系统受到抑制

The changes of expression level of matrix metalloprotease-9 and its inhibitor (TIMP-1) in murine pulmonary fibrosis model

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(d0.05). The concentration of MMP-9 secreted by AMs was gradually decreased on day 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.