Shin Jae-Min, Park Il-Ho, Moon You-Mi, Hong Sung-Moon, Cho Jung-Sun, Um Ji-Young, Lee Heung-Man
Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Guro Hospital, Korea University, Seoul, Korea.
Am J Rhinol Allergy. 2014 May-Jun;28(3):e120-4. doi: 10.2500/ajra.2014.28.4039.
Fibroblast migration is crucial for normal wound repair after sinonasal surgery. Prostaglandin E2 (PGE2) is a potent inhibitor of fibroblast functions including chemotaxis, proliferation, and matrix production. The purpose of this study was to determine whether PGE2 affects the migration of nasal fibroblasts and to investigate the mechanism of action of PGE2 on nasal fibroblasts.
Primary cultures of nasal fibroblasts were established from inferior turbinate samples. Fibroblast migration was evaluated with scratch assays. Reverse-transcription polymerase chain reaction was performed for E prostanoid (EP) 1, EP2, EP3, and EP4 receptors. EP receptor-selective agonists and antagonists were used to evaluate receptor functions. Stimulatory G (Gs) proteins were activated to evaluate mechanisms. Intracellular cyclic adenosine monophosphate (cAMP) levels were measured by ELISA, and fibroblast cytoskeletal structures were visualized with immunocytochemistry.
PGE2 significantly reduced the migration of nasal fibroblasts. Agonists selective for the EP2 and EP4 receptors significantly reduced the nasal fibroblast migration. Antagonists of the EP2 and EP4 receptors inhibited the effect of PGE2 on nasal fibroblast migration. Activation of Gs protein and adenyl cyclase reduced nasal fibroblast migration.
PGE2 inhibited the migration of nasal fibroblasts via the EP2 and EP4 receptors, and this inhibition was mediated by cAMP elevation. Targeting specific EP receptors could offer therapeutic opportunities for conditions such as delayed wound healing after nasal surgery.
成纤维细胞迁移对于鼻科手术后的正常伤口修复至关重要。前列腺素E2(PGE2)是成纤维细胞功能的强效抑制剂,包括趋化性、增殖和基质产生。本研究的目的是确定PGE2是否影响鼻成纤维细胞的迁移,并研究PGE2对鼻成纤维细胞的作用机制。
从下鼻甲样本建立鼻成纤维细胞原代培养物。用划痕试验评估成纤维细胞迁移。对E前列腺素(EP)1、EP2、EP3和EP4受体进行逆转录聚合酶链反应。使用EP受体选择性激动剂和拮抗剂评估受体功能。激活刺激性G(Gs)蛋白以评估机制。通过酶联免疫吸附测定法测量细胞内环磷酸腺苷(cAMP)水平,并用免疫细胞化学观察成纤维细胞细胞骨架结构。
PGE2显著降低鼻成纤维细胞的迁移。对EP2和EP4受体具有选择性的激动剂显著降低鼻成纤维细胞迁移。EP2和EP4受体拮抗剂抑制PGE2对鼻成纤维细胞迁移的作用。Gs蛋白和腺苷酸环化酶的激活降低鼻成纤维细胞迁移。
PGE2通过EP2和EP4受体抑制鼻成纤维细胞的迁移,并且这种抑制是由cAMP升高介导的。靶向特定的EP受体可为诸如鼻科手术后伤口愈合延迟等情况提供治疗机会。