Objective To explore the mechanism of hypoxia on the expressions of airway remodeling-associated factors matrix metalloproteinase-9(MMP-9)and transforming growth factor β1(TGF-β1)in human airway epithelial cells(16HBE)under hypoxia.Methods We cultured 16HBE cells under normoxia or hypoxia (20 mL/L O2)for 6,12 and 24 hours.MMP-9 and TGF-β1with higher expression were pretreated with epidermal growth factor receptor(EGFR)inhibitor AG1478 and hypoxia-inducible factor-1α(HIF-1α)inhibitor Lificiguat (YC-1).Cell survival rate was measured by CCK-8 method.The mRNA levels of MMP-9 and TGF-β1were detected by RT-PCR.The levels of HIF-1α,MMP-9 and TGF-β1protein expressions were measured by Western blot. Results In hypoxia group,the levels of MMP-9 and TGF-β1mRNA as well as protein expressions increased compared with those in the control group(all P＜ 0.05).Pretreatment with AG1478 and YC-1 could inhibit the above-mentioned changes(P＜ 0.05).AG1478 suppressed the high expression of HIF-1α induced by hypoxia. Conclusion Hypoxia can up-regulate the expressions of airway remodeling-associated factors MMP-9 and TGF-β1 via EGFR's induction of HIF-α signaling pathway.%目的 研究低氧刺激对人支气管上皮细胞(16HBE)中基质金属蛋白酶-9(MMP-9)和转化生长因子-β1(TGF-β1)表达的影响及其相关信号机制.方法 将细胞分组,低氧组用20 mL/L O2培养16HBE细胞,培养时间分别为6、12、24 h;对照组于常规条件下培养.选择MMP-9和TGF-β1表达较高的时间组予以表皮生长因子受体(EGFR)抑制剂AG1478、低氧诱导因子(HIF-1α)抑制剂Lificiguat(YC-1)干预.CCK-8法检测各组细胞活力,实时荧光定量 PCR检测MMP-9和TGF-β1的转录水平,Western blot检测 HIF-1α、MMP-9和TGF-β1的蛋白相对表达水平.结果 低氧组MMP-9和TGF-β1转录及蛋白水平均高于对照组(P均＜ 0.05),AG1478、YC-1可抑制低氧引起的上述改变(P＜0.05);AG1478能降低在低氧诱导时高水平表达的 HIF-1α.结论 低氧刺激可能通过EGFR诱导 HIF-1α表达,进而促进气道重塑相关因子MMP-9和TGF-β1的表达.
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