IL-4-induced caveolin-1-containing lipid rafts aggregation contributes to MUC5AC synthesis in bronchial epithelial cells.
IL-4-induced caveolin-1-containing lipid rafts aggregation contributes to MUC5AC synthesis in bronchial epithelial cells.
复制标题
IL-4诱导的含有caveolin-1的脂筏聚集有助于支气管上皮细胞中MUC5AC的合成
DOI：
10.1186/s12931-017-0657-z
复制
发表时间：
2017-09-20
影响因子：
5.8
通讯作者：
Ye H
中科院分区：
文献类型：
--
作者：
Xia Y;Cai PC;Yu F;Xiong L;He XL;Rao SS;Chen F;Yang XP;Ma WL;Ye H
粘液分泌过多是哮喘的一个重要特征。白细胞介素 (IL)-4 是过敏原引起的气道炎症和粘液产生所必需的。 MUC5AC 基因表达受转录因子 NF-κB 调节。细胞内 Ca2+ ([Ca2+]i) 信号是 NF-κB 激活所必需的。瞬时受体电位规范 1 (TRPC1) 通道已被证明有助于某些类型细胞中激动剂刺激的 Ca2+ 流入。然而，哮喘患者中 IL-4、TRPC1 和支气管上皮细胞 (BEC) 粘液过量产生之间的关系尚不清楚。从大鼠大支气管气道中分离BECs并用作细胞模型。为了呈现脂筏、caveolin-1 和 TRPC1 的变化，进行了免疫荧光染色和蔗糖梯度离心。 [Ca2+]i 在加载 Fura-2 后进行测量。通过基于 ELISA 的测定法测量 NF-κB 活性。分别通过实时定量RT-PCR、ELISA分析和免疫荧光染色检测MUC5AC mRNA和蛋白水平。 IL-4 诱导 BEC 中 Ca2+ 内流，而 Ca2+ 内流抑制剂 (2-APB) 可阻断这一过程。 2-APB 还阻止 IL-4 诱导的 MUC5AC 蛋白合成。细胞外 Ca2+ 的消耗导致 IL-4 处理的细胞中 MUC5AC 表达部分下降。 NF-κB 而不是 STAT6 激活介导 IL-4 诱导的 MUC5AC 蛋白合成。然后研究了Ca2+内流的机制。免疫荧光染色和蔗糖梯度离心表明，含有caveolin-1的脂筏聚集参与了BEC中TRPC1激活和Ca2+流入。最后，数据显示，阻断脂筏聚集可以准确地阻止 IL-4 诱导的 Ca2+ 内流、NF-κB 激活和 MUC5AC 合成。我们的结果表明，IL-4诱导的含有caveolin-1的脂筏聚集至少部分有助于BEC中MUC5AC的合成。本文的在线版本 (10.1186/s12931-017-0657-z) 包含补充材料，可供授权用户使用。
Mucus overproduction is an important feature of asthma. Interleukin (IL)-4 is required for allergen-induced airway inflammation and mucus production. MUC5AC gene expression is regulated by transcript factors NF-κB. The intracellular Ca2+ ([Ca2+]i) signal is required for activation of NF-κB. The transient receptor potential canonical 1 (TRPC1) channel has been shown to contribute for agonist-stimulated Ca2+ influx in some types of cells. However, the relationships among IL-4, TRPC1 and mucus overproduction in bronchial epithelial cells (BECs) in asthma are poorly understood. BECs were isolated from large bronchial airway of rats and used as cell model. To present changes of lipid raft, caveolin-1 and TRPC1, immunofluorescence staining and sucrose gradient centrifugation were performed. [Ca2+]i was measured after loading with Fura-2. NF-κB activities were measured by an ELISA-based assay. MUC5AC mRNA and protein levels were detected by real-time quantitative RT-PCR, ELISA analysis and immunofluorescence staining respectively. IL-4 induced Ca2+ influx in BECs, and this was blocked by a Ca2+ influx inhibitor (2-APB). 2-APB also prevented MUC5AC protein synthesis induced by IL-4. Depletion of extracellular Ca2+ resulted in partial decrease in expression of MUC5AC in IL-4 treated cells. NF-κB rather than STAT6 activation mediated IL-4-induced MUC5AC protein synthesis. Then the mechanism of Ca2+ influx was investigated. Immunofluorescence staining and sucrose gradient centrifugation revealed that caveolin-1-containing lipid rafts aggregation was involved in TRPC1 activation and Ca2+ influx in BECs. Lastly, the data revealed that blocking lipid rafts aggregation exactly prevented Ca2+ influx, NF-κB activation and MUC5AC synthesis induced by IL-4. Our results indicate that IL-4-induced caveolin-1-containing lipid rafts aggregation at least partly contributes to MUC5AC synthesis in BECs. The online version of this article (10.1186/s12931-017-0657-z) contains supplementary material, which is available to authorized users.
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DOI：
10.1002/ptr.5334
发表时间：
2015-06
期刊：
Phytotherapy research : PTR
影响因子：
--
作者：
Liu C;Weir D;Busse P;Yang N;Zhou Z;Emala C;Li XM
通讯作者：
Li XM
影响因子：
4
作者：
Alicia, Sampieri;Angelica, Zepeda;Vaca, Luis
通讯作者：
Vaca, Luis
影响因子：
4
作者：
Kim, Cheol Hong;Kim, Kyu-Earn;Song, Kyoung Seob
通讯作者：
Song, Kyoung Seob
影响因子：
3.4
作者：
Lorent, Joseph Helmuth;Levental, Ilya
通讯作者：
Levental, Ilya
影响因子：
15.9
作者：
Laoukili, J;Perret, E;Tournier, F
通讯作者：
Tournier, F