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- 2018
低氧对人支气管上皮细胞气道重塑相关因子表达的影响
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Abstract:
摘要：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20mL/L O2培养16HBE细胞，培养时间分别为6、12、24h；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。
ABSTRACT: 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 (20mL/L O2) for 6, 12 and 24 hours. MMP-9 and TGF-β1 with 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-β1 were detected by RT-PCR. The levels of HIF-1α, MMP-9 and TGF-β1 protein expressions were measured by Western blot. Results In hypoxia group, the levels of MMP-9 and TGF-β1 mRNA 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
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