肿瘤坏死因子-α、刀豆素A及机械损伤刺激人支气管上皮细胞系H292表达基质金属蛋白酶-9

TNFα, ConA and mechanical injury stimulate matrix metalloproteinase-9 expression in human bronchial epithelial cell line h292

目的 :探讨人支气管上皮细胞在受到肿瘤坏死因子 (TNF -α)、刀豆素A(ConA)及机械损伤刺激时基质金属蛋白酶 - 9(MMP - 9)的表达情况。方法 :采用逆转录聚合酶链式反应 (RT -PCR)和金属蛋白酶功能测定法(gelatinzymograhpy) ,观察肿瘤坏死因子 (TNF -α)、刀豆素A(ConA)及机械损伤刺激人支气管上皮细胞系H2 92后 4个时点 (2、6、12h及 2 4h)MMP - 9表达和分泌的动态变化。结果 :TNF -α、ConA及机械损伤刺激均可诱导人支气管上皮细胞系H2 92表达和分泌MMP - 9,无刺激时则不表达 ;单纯机械刺激后 2h ,MMP - 9mRNA开始表达 ,12h达高峰 ,2 4h仍有表达 ,但有所降低 ;ConA与机械损伤联合刺激时 ,MMP - 9表达量明显高于其它各组。同时显示 ,仅在刺激后 2 4h表达MMP - 9活性 ,且以ConA与机械损伤联合刺激组MMP - 9活性为最高。结论 :人支气管上皮细胞在受到TNF -α、ConA及机械损伤刺激时 ,可表达分泌MMP - 9。

AIM: To study the expression of matrix metalloproteinase-9 (MMP-9) in human bronchial epithelial cell treated with TNF-α, ConA and mechanical injury. METHODS: RT-PCR and gelatin zymography were performed to observe the dynamic expression of matrix metalloproteinase-9(MMP-9) in human bronchial epithelial cell line H292 at different timepoint(2 h, 6 h,12 h, 24 h) after stimulation with TNF-α, ConA and mechanical injury. RESULTS: Human bronchial epithelial cell H292 expressed and secreted matrix metalloproteinase-9 in the presence of TNF-α, ConA and mechanical injury. On stimulation with mechanical injury alone, MMP-9 was initially expressed at 2 h, peaked at 12 h, and decreased at 24 h. Furthermore, on stimulation with combination of ConA and mechanical injury, the MMP-9 mRNA expression was the highest among in all the groups. Zymography show that the MMP-9 activity appeared just at 24 h timepoint, and was highest at the group of combination of ConA and mechanical injury. CONCLUTION: Human bronchial epithelial cell expressed and secreted MMP-9 when treated with TNF-α, ConA and mechanical injury.