Prostaglandin E2 Promotes Wound-Induced Migration of Intestinal Subepithelial Myofibroblasts via EP2, EP3, and EP4 Prostanoid Receptor Activation
Prostaglandin E2 Promotes Wound-Induced Migration of Intestinal Subepithelial Myofibroblasts via EP2, EP3, and EP4 Prostanoid Receptor Activation
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DOI：
10.1124/jpet.111.189845
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发表时间：
2012-03-01
影响因子：
3.5
通讯作者：
Ozaki, Hiroshi
中科院分区：
文献类型：
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作者：
Iwanaga, Koichi;Okada, Muneyoshi;Ozaki, Hiroshi
肠上皮下肌成纤维细胞（ISMF）是存在于整个肠的上皮下区域中的间充质细胞。当肠道受损时，ISMF的迁移和有丝分裂反应对于伤口闭合至关重要。然而，其作用机制仍不清楚。我们研究了环氧合酶（考克斯）及其代谢产物前列腺素E-2（PGE（2））在牛ISMF创伤修复过程中的作用。在细胞培养物中的ISMF层中的机械划痕作用在事件发生后1和6小时升高了考克斯-2 mRNA表达和PGE（2）分泌水平。24小时后，ISMF迁移到并减少了划痕部位周围的受伤区域。用考克斯-1/2抑制剂吲哚美辛、考克斯-2抑制剂3-（4-甲磺酰基苯基）-4-苯基-5-三氟甲基异恶唑（CAY 10404）或E前列腺素类受体2至4（EP 2-EP 4）拮抗剂治疗显著抑制伤口修复。相反，用PGE（2）或EP 2、EP 3或EP 4受体激动剂治疗可逆转吲哚美辛对伤口闭合的抑制作用，但不能逆转EP 1受体激动剂。虽然EP 2至EP 4刺激不影响ISMF增殖，但在transwell细胞迁移测定中它确实刺激ISMF迁移。值得注意的是，由EP 2和EP 4刺激的细胞迁移被酪氨酸激酶受体抑制剂染料木黄酮以及（Z）-3-[2，4-二甲基-5-（2-氧代-1，2-二氢-吲哚-3-亚甲基）-1H-吡咯-3-基]-丙酸（SU 6668）抑制。然而，EP 3刺激的细胞迁移不受影响。逆转录-聚合酶链反应显示EP 2或EP 4刺激提高了成纤维细胞生长因子-2的mRNA表达水平，从而刺激ISMF迁移。总之，考克斯-2依赖性PGE（2）分泌通过ISMF促进伤口愈合。PGE（2）-EP 3信号可能直接刺激ISMF迁移。PGE（2）-EP 2/4信号通过提高生长因子分泌水平间接刺激ISMF迁移。
Intestinal subepithelial myofibroblasts (ISMFs) are mesenchymal cells that reside in the subepithelial region throughout the intestine. When the intestine is damaged, the migratory and mitotic responses of ISMFs are crucial for wound closure. However, their mechanism of action remains unknown. We have investigated the role of cyclooxygenase (COX) and its metabolite prostaglandin E-2 (PGE(2)) in the wound repair process of bovine ISMFs. The action of a mechanical scratch in a layer of ISMFs in cell culture elevated the levels of both COX-2 mRNA expression and PGE(2) secretion 1 and 6 h after the event. After 24 h ISMFs had migrated to and reduced the wounded area around the site of the scratch. Treatment with the COX-1/2 inhibitor indomethacin, the COX-2 inhibitor 3-(4-methylsulphonylphenyl)-4-phenyl-5-trifluoromethylisoxazole (CAY10404), or E prostanoid receptor 2 to 4 (EP2-EP4) antagonists significantly inhibited wound repair. Conversely, inhibition of wound closure by indomethicin was reversed by treatment with PGE(2) or agonists of the receptors EP2, EP3, or EP4 but not of EP1. Although EP2 to EP4 stimulation did not influence ISMF proliferation, it did stimulate ISMF migration in the transwell cell migration assay. It is noteworthy that cell migration stimulated by EP2 and EP4 was inhibited by the tyrosine kinase receptor inhibitor genistein and also by (Z)-3-[2,4-dimethyl-5-(2-oxo-1,2-dihydro- indol-3-ylidenemethyl)-1H-pyrrol-3-yl]-propionic acid (SU6668). However, cell migration stimulated by EP3 was unaffected. Reverse transcription-polymerase chain reaction showed EP2 or EP4 stimulation elevated the level of mRNA expression for fibroblast growth factor-2, which stimulates ISMF migration. Collectively, COX-2-dependent PGE(2) secretion promotes wound healing by ISMFs. PGE(2)-EP3 signaling may directly stimulate ISMF migration. PGE(2)-EP2/4 signaling indirectly stimulates ISMF migration by elevating the level of growth factor secretion.