Leukotriene E4 induces MUC5AC release from human airway epithelial NCI-H292 cells.
Leukotriene E4 induces MUC5AC release from human airway epithelial NCI-H292 cells.
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DOI：
10.1016/j.alit.2014.11.002
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发表时间：
2015-04
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影响因子：
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通讯作者：
H. Shirasaki;E. Kanaizumi;N. Seki;T. Himi
中科院分区：
文献类型：
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作者：
H. Shirasaki;E. Kanaizumi;N. Seki;T. Himi
背景气道上皮粘蛋白的高分泌是变应性气道疾病的重要特征。在3种半胱氨酰白三烯（CysLTs; LTC 4LTD 4和LTE 4）中，只有LTE 4足够稳定，可在细胞外液中检测到。然而，LTE 4很少受到关注，因为它与CysLT 1和CysLT 2受体结合不良，因此，LTE 4对粘液分泌的影响知之甚少。最近，研究集中在P2 Y12受体作为LTE 4的潜在受体，因为该受体是LTE 4介导的肺部炎症所必需的。在我们前期的研究中，我们证实了P2 Y12受体在人气道上皮细胞中的表达。方法用LTE 4（0.01 ~ 1 μM）刺激NCI-H292细胞24 h，观察LTE 4对气道上皮细胞粘液分泌的影响。释放和生产的MUC 5AC蛋白，凝胶形成粘蛋白，进行了评价与酶联免疫吸附试验。ResultsWestern blot分析显示，NCI-H292细胞表达P2 Y12受体蛋白。LTE 4以剂量依赖性方式显著诱导MUC 5AC粘蛋白的释放。Th 2细胞因子如IL-4（10 ng/mL）和IL-13（10 ng/mL）加速LTE 4诱导的MUC 5AC蛋白的释放。P2 Y12受体拮抗剂MRS 2935可部分抑制LTE 4诱导的气道MUC 5AC蛋白的释放。CysLT 1受体拮抗剂MK 571对LTE 4诱导的MUC 5AC蛋白的释放无影响。结论LTE 4可能通过激活P2 Y12受体在变应性粘液分泌中发挥重要作用。
BackgroundHypersecretion of mucin in the airway epithelium is an important feature of allergic airway diseases. Of the 3 cysteinyl leukotrienes (CysLTs; LTC4LTD4and LTE4), only LTE4is sufficiently stable to be detectable in extracellular fluids. However, LTE4has received little attention because it binds poorly to the CysLT1and CysLT2receptors; therefore, little is known about the effects of LTE4on mucous secretion. Recently, studies have focused on the P2Y12receptor as a potential receptor for LTE4, because this receptor is required for LTE4-mediated pulmonary inflammation. In our previous study, we confirmed the expression of P2Y12receptor in human airway epithelial cells. To clarify the roles of LTE4in airway epithelial cells, we investigated mucus secretion by LTE4in vitro.MethodsConfluent NCI-H292 cells were stimulated with LTE4(0.01–1 μM) for 24 h. The release and production of MUC5AC protein, a gel-forming mucin, were evaluated with an enzyme-linked immunosorbent assay.ResultsWestern blot analysis revealed that NCI-H292 cells expressed P2Y12receptor protein. LTE4significantly induced the release of MUC5AC mucin in a dose-dependent manner. Th2 cytokines such as IL-4 (10 ng/mL) and IL-13 (10 ng/mL) accelerated the LTE4-induced release of MUC5AC protein. MRS2935, a P2Y12receptor antagonist, partially inhibited the LTE4-induced release of MUC5AC protein in the airway. In contrast, MK571, a CysLT1receptor antagonist, did not affect the release of MUC5AC protein elicited by LTE4.ConclusionsThese results suggest that LTE4may play some important roles in allergic mucus secretion partially via activation of P2Y12receptor.