Submucosal connective tissue-type mast cells contribute to the production of lysophosphatidic acid (LPA) in the gastrointestinal tract through the secretion of autotaxin (ATX)/lysophospholipase D (lysoPLD)
Submucosal connective tissue-type mast cells contribute to the production of lysophosphatidic acid (LPA) in the gastrointestinal tract through the secretion of autotaxin (ATX)/lysophospholipase D (lysoPLD)
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DOI：
10.1007/s00428-007-0425-4
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发表时间：
2007-07-01
期刊：
影响因子：
3.5
通讯作者：
Nagawa, Hirokazu
中科院分区：
文献类型：
--
作者：
Mori, Ken;Kitayama, Joji;Nagawa, Hirokazu
溶血磷脂酸（LPA）参与广泛的生物活性，包括伤口愈合和癌症转移。自分泌运动因子（ATX）最初作为肿瘤细胞运动刺激因子从黑素瘤上清液中分离，已显示与溶血磷脂酶D（lysoPLD）分子上相同，溶血磷脂酶D是LPA产生中的主要酶。虽然已知ATX/lysoPLD在正常人体组织中广泛表达，但ATX产生细胞的确切分布尚未得到充分研究。在这项研究中，我们评估ATX/lysoPLD的表达，免疫组化染色使用大鼠抗ATX单克隆抗体在人胃肠道，发现粘膜下肥大细胞（MC）高度表达这种酶。这通过使用针对类胰蛋白酶和糜蛋白酶的mAb的免疫荧光双染色来证实。然后，我们通过免疫磁性方法使用CD 117-微珠从人胃组织中分离MC，并显示CD 203 c阳性MC的亚群在流式细胞术上显示细胞内ATX/lysoPLD阳性染色。这通过分离的细胞的Western印迹证实。Western blot检测到人MC培养上清中ATX/lysoPLD的释放。我们的数据表明，粘膜下MC在胃肠道的病理生理学的各个方面发挥重要作用，通过生产ATX/lysoPLD局部提供生物活性LPA。
Lysophosphatidic acid (LPA) is involved in a broad spectrum of biological activities, including wound healing and cancer metastasis. Autotaxin (ATX), originally isolated from a melanoma supernatant as a tumor cell motility-stimulating factor, has been shown to be molecularly identical to lysophospholipase D (lysoPLD), which is the main enzyme in the production of LPA. Although ATX/lysoPLD is known to be widely expressed in normal human tissues, the exact distribution of ATX-producing cells has not been fully investigated. In this study, we evaluated ATX/lysoPLD expression by immunohistochemical staining using a rat anti-ATX mAb in the human gastrointestinal tract and found that submucosal mast cells (MC) highly expressed this enzyme. This was confirmed by immunofluo rescent double staining using mAbs to tryptase and chymase. Then, we isolated MC from human gastric tissue by an immunomagnetic method using CD 117-microbeads and showed that a subpopulation of CD203c-positive MC showed positive staining for intracellular ATX/lysoPLD on flowcytometry. This was confirmed by Western blotting of the isolated cells. Moreover, a significant level of ATX/lysoPLD release could be detected in the culture supernatants of human MC by Western blot analysis. Our data suggest that submucosal MC play significant roles in various aspects of pathophysiology in the gastrointestinal tract by locally providing bioactive LPA through the production of ATX/lysoPLD.