Purinergic P2Y1 receptor signaling mediates wound stimuli-induced cyclooxygenase-2 expression in intestinal subepithelial myofibroblasts
Purinergic P2Y1 receptor signaling mediates wound stimuli-induced cyclooxygenase-2 expression in intestinal subepithelial myofibroblasts
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嘌呤能 P2Y1 受体信号介导伤口刺激诱导的肠上皮下肌成纤维细胞中环氧合酶 2 的表达
DOI：
10.1016/j.ejphar.2013.01.025
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发表时间：
2013
期刊：
影响因子：
5
通讯作者：
Ozaki H
中科院分区：
文献类型：
--
作者：
Iwanaga K;Murata T;Hori M;Ozaki H
肠上皮下肌成纤维细胞（ISMF）对于针对炎症刺激的屏障形成至关重要。物理损伤诱导环氧合酶-2（考克斯-2）表达，其通过ISMF加速伤口愈合。然而，考克斯-2诱导的机制仍不清楚。身体受损的细胞释放ATP。在这里，我们调查的作用，ATP-嘌呤信号在创伤诱导考克斯-2诱导ISMF。伤后24小时，牛ISMF已迁移到并关闭受伤区域。一种考克斯抑制剂，吲哚美辛或嘌呤能P2受体拮抗剂，苏拉明，抑制伤口愈合。然而，使用消炎痛进行额外治疗并不影响苏拉明治疗的ISMF的伤口愈合。RT-PCR显示伤后2 h考克斯-2 mRNA表达增加，苏拉明可抑制COX-2 mRNA表达。这些结果表明，ATP介导创伤诱导的考克斯-2升高。我们接下来评估了各种嘌呤能受体在考克斯-2诱导中的作用。ATP类似物ATPγS和嘌呤能P2 Y1、11-13受体激动剂ADP是增加考克斯-2表达的试验药物。ATPγ S诱导的考克斯-2 mRNA表达可被苏拉明或嘌呤能P2 Xs、P2 Y1、4、6和13受体拮抗剂PPADS抑制。这些数据表明Gq偶联嘌呤能P2 Y1受体或Gi偶联嘌呤能P2 Y13受体参与考克斯-2诱导。Gq蛋白下游信号磷脂酶C抑制剂U 73122可抑制考克斯-2 mRNA的表达。然而，百日咳毒素，一种Giinhibitor，没有显示抑制作用。p38 MAPK和PKC抑制剂可抑制ATPγ S诱导的考克斯-2 mRNA表达。总之，嘌呤能P2 Y1受体信号通过p38 MAPK和PKC途径介导ISMF中创伤诱导的考克斯-2表达。
Intestinal subepithelial myofibroblasts (ISMFs) are crucial for barrier formation against inflammatory stimuli. Physical injury induces cyclooxygenase-2 (COX-2) expression, which accelerates wound healing by ISMFs. However, the mechanism of COX-2 induction remains unclear. Physically damaged cells release ATP. Here, we investigate the role of ATP-purinergic signaling in wound-induced COX-2 induction in ISMFs. By 24h post-injury, bovine ISMFs had migrated to and closed the wounded area. A COX inhibitor, indomethacin or a purinergic P2 receptor antagonist, suramin, inhibited wound healing. However, additional treatment with indomethacin did not influence wound healing in suramin-treated ISMFs. RT-PCR showed an increase in COX-2 mRNA expression 2h post-injury, which was inhibited by suramin. These results suggest that ATP mediates wound-induced COX-2 elevation. We next assessed the contribution of various purinergic receptors in COX-2 induction. An ATP analog, ATPγS and a purinergic P2Y1, 11–13 receptors agonist, ADP, were among the agents tested which increased COX-2 expression. ATPγS-induced COX-2 mRNA expression was suppressed by suramin or a purinergic P2Xs, P2Y1, 4, 6, and 13 receptors antagonist, PPADS. These data suggest the involvement of Gq-coupled purinergic P2Y1 receptor or Gi-coupled purinergic P2Y13 receptor in COX-2 induction. U73122, an inhibitor of phospholipase C, which is a downstream signal of Gqprotein, showed suppression of COX-2 mRNA expression. However, pertussis toxin, a Giinhibitor, did not show suppression. We also revealed that inhibitors of p38 MAPK and PKC inhibited ATPγS-induced COX-2 mRNA expression. Collectively, purinergic P2Y1 receptor signaling mediates wound-induced COX-2 expression through p38 MAPK and PKC pathways in ISMFs.