中文摘要：

目的观察支原体巨噬细胞活化脂肽-2（MALP-2）诱导人气道上皮细胞分泌粘蛋白MUC5AC的分子机制。方法体外培养人气道上皮细胞NCI-H292,分别采用0、0.1、1.0和5.0μg/m L MALP-2刺激NCI-H292细胞24 h,采用酶联免疫吸附测定（ELISA）检测培养上清中MUC5AC和基质金属蛋白酶9（MMP-9）的含量;Western blot检测表皮生长因子受体（EGFR）磷酸化水平。同时采用EGFR抑制剂AG-1478或MMP-9抑制剂（MMP-9 Inhibitor I）处理细胞,观察其对MUC5AC分泌的影响。结果 NCI-H 292细胞未刺激时,MUC 5 AC以及MMP-9分泌水平极低。当给予0.1~5μg/m L MALP-2作用18 h后,MUC 5 AC的分泌水平显著增高。此外,5μg/m L MALP-2作用NCI-H 292细胞1 h后可诱导EGFR磷酸化。采用AG-1478预处理细胞1 h后,MMP-9及MUC 5 AC分泌水平明显减少,同时,采用10 nmol/L MMP-9抑制剂处理后也能下调MUC 5 AC水平。结论支原体MALP-2经EGFR/MMP-9诱导气道上皮细胞分泌MUC5AC。

英文摘要：

Objective To investigate the molecular mechanism of the Macrophage-activating lipopeptide-2 （MALP-2） on secretion of MUC5AC in human airway epithelial cells. Methods The airway epithelial cell line NCI-H 292 was cultured in vitro and stimulated with 0, 0.1,1.0 and 5.0 ktg/ mL of MALP-2 for 24 h. Secretion of MUC5AC and matrix metalloprotein-9 （MMP-9） in the supernatant were detected by ELISA; Phosphorylation of epithelial growth factor receptor （EGFR） was measured by Western blot. To observe the effect of EGFR and MMP-9 on the mediation of MUC5AC secretion, specific inhibitor AG-1478 and MMP-9 Inhibitor I was used before MALP-2 stimulation. Results The secretion level of MUC5AC and MMP-9 was very low in untreated cells. 0.1-51xg/mL of MALP-2 incubation for 18h significantly upregulated MUC 5 AC secretion. In addition, 5~tg/mL MALP-2 could induce EGFR phosphorylation after lh of incubation. Treatment of AG-1478, an inhibitor of EGFR, significantly abrogated MALP-2-induced MMP-9 and MUC 5 AC secretion. Furthermore, 10nmol/L MMP-9 inhibitor could also inhibit MUC 5 AC production. Conclusion Mycoplasma MALP-2 induces MUC 5 AC secretion via the EGFR/MMP-9 pathways in human airway epithelial cells