Prostaglandin E2-Transporting Pathway and Its Roles via EP2/EP4 in Cultured Human Dental Pulp
Prostaglandin E2-Transporting Pathway and Its Roles via EP2/EP4 in Cultured Human Dental Pulp
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培养人牙髓中前列腺素 E2 转运途径及其通过 EP2/EP4 的作用
DOI：
10.1016/j.joen.2023.01.009
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发表时间：
2023
影响因子：
4.2
通讯作者：
Noiri Yuichiro
中科院分区：
文献类型：
--
作者：
Ohkura Naoto;Yoshiba Kunihiko;Yoshiba Nagako;Oda Yohei;Edanami Naoki;Ohshima Hayato;Takenaka Shoji;Okiji Takashi;Noiri Yuichiro
前列腺素E2（PGE 2）通过细胞内合成、细胞外转运和受体结合等转运途径发挥生物学作用。本研究的目的是确定在人牙髓中的PGE 2转运途径的组件的定位和探索的相关性PGE 2受体（EP 2/EP 4）的血管生成和dentinogenes.MethodsProtein定位的微粒体PGE 2（mPGES）合成酶，PGE 2转运蛋白（多药耐药相关蛋白4 [MRP 4]和前列腺素转运蛋白[PGT]），和EP 2/EP 4使用双重免疫荧光染色进行了分析。将来自人第三磨牙的牙齿切片与或不与布他前列素（EP 2激动剂）或利文前列素（EP 4激动剂）一起培养1周。结果MRP 4、PGT与mPGES、EP 2/EP 4在成牙本质细胞和内皮细胞中共定位。此外，MRP 4与mPGES和EP 4共定位于表达人白细胞抗原DR的树突状细胞中。在牙片培养中，EP 2/EP 4激动剂诱导丝状伪足的数量和长度以及血管生成标记物的mRNA表达显著增加（血管内皮生长因子和成纤维细胞生长因子-2）和成牙本质细胞分化标志物结论前列腺素E2产生酶（mPGES）、转运蛋白、牙本质涎磷蛋白和胶原蛋白1在牙本质中的表达与前列腺素E2产生酶（mPGES）的表达密切相关。（MRP 4和PGT）和PGE 2特异性受体（EP 2/EP 4）在人牙髓的各种细胞成分中进行免疫定位。EP 2/EP 4激动剂促进内皮细胞丝状伪足的产生，上调血管生成和成牙本质细胞分化相关基因，表明PGE 2与EP 2/EP 4结合与血管生成和牙本质生成反应相关。
IntroductionProstaglandin E2(PGE2) exerts biological actions through its transport pathway involving intracellular synthesis, extracellular transport, and receptor binding. This study aimed to determine the localization of the components of the PGE2-transporting pathway in human dental pulp and explore the relevance of PGE2receptors (EP2/EP4) to angiogenesis and dentinogenesis.MethodsProtein localization of microsomal PGE2(mPGES)synthase, PGE2transporters (multidrug resistance-associated protein-4 [MRP4] and prostaglandin transporter [PGT]), and EP2/EP4 was analyzed using double immunofluorescence staining. Tooth slices from human third molars were cultured with or without butaprost (EP2 agonist) or rivenprost (EP4 agonist) for 1 week. Morphometric analysis of endothelial cell filopodia was performed to evaluate angiogenesis, and real-time polymerase chain reaction was performed to evaluate angiogenesis and odontoblast differentiation markers.ResultsMRP4 and PGT were colocalized with mPGES and EP2/EP4 in odontoblasts and endothelial cells. Furthermore, MRP4 was colocalized with mPGES and EP4 in human leukocyte antigen-DR-expressing dendritic cells. In the tooth slice culture, EP2/EP4 agonists induced significant increases in the number and length of filopodia and mRNA expression of angiogenesis markers (vascular endothelial growth factor and fibroblast growth factor-2) and odontoblast differentiation markers (dentin sialophosphoprotein and collagen type 1).ConclusionsPGE2-producing enzyme (mPGES), transporters (MRP4 and PGT), and PGE2-specific receptors (EP2/EP4) were immunolocalized in various cellular components of the human dental pulp. EP2/EP4 agonists promoted endothelial cell filopodia generation and upregulated angiogenesis- and odontoblast differentiation-related genes, suggesting that PGE2binding to EP2/EP4 is associated with angiogenic and dentinogenic responses.