分泌型磷脂酶A2是由纤毛细胞分泌而增加的粘蛋白与类花生四烯酸由杯状细胞分泌
2016/04/28
摘要
背景：分泌型的磷脂酶A2（secretory phospholipases A2, sPLA2）在重症哮喘患者气道中的分泌是增加的，其可以启动类花生四烯酸类的生物合成，并诱导粘液高分泌。我们用IL-13活化的高度富集的杯状细胞及其分化的人支气管上皮细胞（纤毛细胞富集）培养来评估纤毛细胞和杯状细胞对气道 sPLA2合成与应答的相对作用。我们希望能发现人气道上皮细胞中sPLA2以及白三烯的主要来源。
方法：我们使用无肺部疾病受试者的支气管上皮细胞来分化成为纤毛细胞富集或杯状细胞富集的细胞表型。 sPLA2、半胱氨酸白三烯（cysteinyl leukotrienes，cysLTs）与气道粘蛋白mRNA 及其蛋白质的合成通过实时-聚合酶联反应（RT-PCR）与酶联免疫吸附测定（ELISA）来检测，此外，粘蛋白与sPLA2在特殊细胞类型的定位通过荧光共聚焦显微镜来证实。
结果：IIa, V, 和 X 型sPLA2的信使RNA（mRNA） 在富集纤毛细胞的细胞表型中增多，而不是在富集杯状细胞的细胞表型增多（P< .001）。 sPLA2是由纤毛细胞的顶（空气）层而不由杯状细胞分泌(P<.001)。V型sPLA2在纤毛富集细胞而不是杯状富集细胞内的免疫染色呈强阳性。sPLA2通过杯状富集细胞而不是纤毛富集细胞来介导释放cysLTs，且V型sPLA2的作用效果最强(P< .05)。V型sPLA2增加杯状富集细胞的粘蛋白分泌，这种效应可以被脂氧化酶抑制剂或环氧化酶抑制剂所抑制(P< .02)。
结论： sPLA2由纤毛细胞分泌，且它似乎能诱导杯状细胞分泌粘蛋白与cysLT，这强烈支持气道杯状细胞是一种促炎效应细胞。
Secretory Phospholipases A2 Are Secreted From Ciliated Cells and Increase Mucin and Eicosanoid Secretion From Goblet Cells
Tanabe T, Shimokawaji T, Kanoh S, Rubin BK.
Chest. 2015;147(6):1599-609.
ABSTRACT
BACKGROUND: Secretory phospholipases A2 (sPLA2) initiate the biosynthesis of eicosanoids, are increased in the airways of people with severe asthma, and induce mucin hypersecretion. We used IL-13-transformed, highly enriched goblet cells and differentiated (ciliary cell-enriched) human bronchial epithelial cell culture to evaluate the relative contribution of ciliated and goblet cells to airway sPLA2 generation and response. We wished to determine the primary source(s) of sPLA2 and leukotrienes in human airway epithelial cells.
METHODS: Human bronchial epithelial cells from subjects without lung disease were differentiated to a ciliated-enriched or goblet-enriched cell phenotype. Synthesis of sPLA2, cysteinyl leukotrienes (cysLTs), and airway mucin messenger RNA and protein was measured by realtime -polymerase chain reaction and an enzyme-linked immunosorbent assay, and the localization of mucin and sPLA2 to specific cells types was confirmed by confocal microscopy.
RESULTS: sPLA2 group IIa, V, and X messenger RNA expression was increased in ciliated-enriched cells (P< .001) but not in goblet-enriched cells. sPLA2 were secreted from the apical (air) side of ciliated-enriched cells but not goblet-enriched cells (P<.001). Immunostaining of sPLA2 V was strongly positive in ciliated-enriched cells but not in goblet-enriched cells. sPLA2 released cysLTs from goblet-enriched cells but not from ciliated-enriched cells, and this result was greatest with sPLA2 V (P< .05). sPLA2 V increased goblet-enriched cell mucin secretion, which was inhibited by inhibitors of lipoxygenase or cyclooxygenase (P< .02).
CONCLUSIONS: sPLA2 are secreted from ciliated cells and appear to induce mucin and cysLT secretion from goblet cells, strongly suggesting that airway goblet cells are proinflammatory effector cells.
背景：分泌型的磷脂酶A2（secretory phospholipases A2, sPLA2）在重症哮喘患者气道中的分泌是增加的，其可以启动类花生四烯酸类的生物合成，并诱导粘液高分泌。我们用IL-13活化的高度富集的杯状细胞及其分化的人支气管上皮细胞（纤毛细胞富集）培养来评估纤毛细胞和杯状细胞对气道 sPLA2合成与应答的相对作用。我们希望能发现人气道上皮细胞中sPLA2以及白三烯的主要来源。
方法：我们使用无肺部疾病受试者的支气管上皮细胞来分化成为纤毛细胞富集或杯状细胞富集的细胞表型。 sPLA2、半胱氨酸白三烯（cysteinyl leukotrienes，cysLTs）与气道粘蛋白mRNA 及其蛋白质的合成通过实时-聚合酶联反应（RT-PCR）与酶联免疫吸附测定（ELISA）来检测，此外，粘蛋白与sPLA2在特殊细胞类型的定位通过荧光共聚焦显微镜来证实。
结果：IIa, V, 和 X 型sPLA2的信使RNA（mRNA） 在富集纤毛细胞的细胞表型中增多，而不是在富集杯状细胞的细胞表型增多（P< .001）。 sPLA2是由纤毛细胞的顶（空气）层而不由杯状细胞分泌(P<.001)。V型sPLA2在纤毛富集细胞而不是杯状富集细胞内的免疫染色呈强阳性。sPLA2通过杯状富集细胞而不是纤毛富集细胞来介导释放cysLTs，且V型sPLA2的作用效果最强(P< .05)。V型sPLA2增加杯状富集细胞的粘蛋白分泌，这种效应可以被脂氧化酶抑制剂或环氧化酶抑制剂所抑制(P< .02)。
结论： sPLA2由纤毛细胞分泌，且它似乎能诱导杯状细胞分泌粘蛋白与cysLT，这强烈支持气道杯状细胞是一种促炎效应细胞。
(王刚 张红萍 张欣 四川大学华西医院中西医结合科呼吸病组 610041 摘译)
（Chest. 2015;147(6):1599-609.）
（Chest. 2015;147(6):1599-609.）
Secretory Phospholipases A2 Are Secreted From Ciliated Cells and Increase Mucin and Eicosanoid Secretion From Goblet Cells
Tanabe T, Shimokawaji T, Kanoh S, Rubin BK.
Chest. 2015;147(6):1599-609.
ABSTRACT
BACKGROUND: Secretory phospholipases A2 (sPLA2) initiate the biosynthesis of eicosanoids, are increased in the airways of people with severe asthma, and induce mucin hypersecretion. We used IL-13-transformed, highly enriched goblet cells and differentiated (ciliary cell-enriched) human bronchial epithelial cell culture to evaluate the relative contribution of ciliated and goblet cells to airway sPLA2 generation and response. We wished to determine the primary source(s) of sPLA2 and leukotrienes in human airway epithelial cells.
METHODS: Human bronchial epithelial cells from subjects without lung disease were differentiated to a ciliated-enriched or goblet-enriched cell phenotype. Synthesis of sPLA2, cysteinyl leukotrienes (cysLTs), and airway mucin messenger RNA and protein was measured by realtime -polymerase chain reaction and an enzyme-linked immunosorbent assay, and the localization of mucin and sPLA2 to specific cells types was confirmed by confocal microscopy.
RESULTS: sPLA2 group IIa, V, and X messenger RNA expression was increased in ciliated-enriched cells (P< .001) but not in goblet-enriched cells. sPLA2 were secreted from the apical (air) side of ciliated-enriched cells but not goblet-enriched cells (P<.001). Immunostaining of sPLA2 V was strongly positive in ciliated-enriched cells but not in goblet-enriched cells. sPLA2 released cysLTs from goblet-enriched cells but not from ciliated-enriched cells, and this result was greatest with sPLA2 V (P< .05). sPLA2 V increased goblet-enriched cell mucin secretion, which was inhibited by inhibitors of lipoxygenase or cyclooxygenase (P< .02).
CONCLUSIONS: sPLA2 are secreted from ciliated cells and appear to induce mucin and cysLT secretion from goblet cells, strongly suggesting that airway goblet cells are proinflammatory effector cells.
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