中国循证医学杂志

中国循证医学杂志

莱菔硫烷对气道黏蛋白 MUC5AC 表达的调控作用

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目的 研究莱菔硫烷(sulforaphane,SFN)对气道黏蛋白 5AC(mucin 5AC,MUC5AC)表达的调控作用并探讨可能的机制。 方法 体外培养人气道上皮细胞A549细胞,分别使用卟啉醇肉豆蔻酸乙酸酯(PMA)和过氧化氢(H2O2)刺激 A549 细胞诱导黏液高分泌模型,使用核因子相关因子 2(Nrf2)信号通路的强诱导剂莱菔硫烷为干预因素,实验分为空白对照组(不加任何刺激)、黏液高表达组(PMA 组和 H2O2 组)及莱菔硫烷干预组(SFN+PMA 组;SFN+H2O2 组)。莱菔硫烷预先处理 30 分钟后分别予 PMA、H2O2 干预 24 小时,分别采用 Real time PCR 和 Western blot 方法检测 A549 细胞 MUC5AC、Nrf2 及其下游抗氧化基因血红素加氧酶 1(HO-1)、醌氧化还原酶 1(NQO1)和谷氨酰半胱氨酸合成酶催化亚单位(GCLC)的 mRNA 含量及蛋白表达水平。 结果 与阴性对照组相比,PMA 组和 H2O2组 MUC5AC mRNA 和蛋白水平显著升高(P 均<0.05)。给予莱菔硫烷处理后,MUC5AC 的表达明显降低(P<0.05),而 Nrf2 及其下游抗氧化基因 HO-1 表达水平较对照组和黏液高表达模型组升高(P 均<0.05)。 结论 莱菔硫烷明显抑制 PMA 及 H2O2 诱导的气道黏蛋白 MUC5AC 基因转录及蛋白合成分泌,可能通过 Nrf2/HO-1 信号通路参与调控气道黏液高分泌。

Objective To explore the role of sulforaphane (SFN) on the regulation of airway mucin 5AC mucin expression in the airway epithelial cell line (A549), and the underlying mechanism. Methods The experiments were performed in culture of PMA and H2O2 induced A549 in vitro. A549 cells divided into three groups: the control group, the model group (PMA and H2O2) and the intervention groups (SFN+PMA; SFN+H2O2). The intervention group's cells were pretreated with SFN for 30 mins before exposure to stimuli (PMA or H2O2). The MUC5AC, Nrf2 and the antioxidant gene HO-1, NQO1, GCLC mRNA levels were analyzed by real time-PCR, and protein production was assayed by western blot. Results Compared with the control group, expression of MUC5AC mucin was increased after being stimulated by PMA or H2O2 (P<0.05), but it was significantly inhibited by SFN (P<0.05). SFN induced the expression of Nrf2 gene and the antioxidant gene HO-1 (compared with the control and model groups,P<0.05). Conclusion Sulforaphane involves the airway mucous hypersecretion induced by PMA and H2O2, and Nrf2/HO-1 signaling pathway may play an important role in mucin MUC5AC regulation.

关键词: 莱菔硫烷; 黏蛋白; MUC5AC; Nrf2 信号通路; 血红素加氧酶-1

Key words: Sulforaphane; Mucus; MUC5AC; Nrf2 signaling pathway; Heme oxygenase-1

引用本文: 林晓萍, 曾奕明. 莱菔硫烷对气道黏蛋白 MUC5AC 表达的调控作用. 中国循证医学杂志, 2017, 17(1): 26-32. doi: 10.7507/1672-2531.201605024 复制

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