Ohkura Naoto, Yoshiba Kunihiko, Yoshiba Nagako, Oda Yohei, Edanami Naoki, Ohshima Hayato, Takenaka Shoji, Okiji Takashi, Noiri Yuichiro
Division of Cariology, Operative Dentistry and Endodontics, Department of Oral Health Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Division of Oral Science for Health Promotion, Department of Oral Health and Welfare, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
J Endod. 2023 Apr;49(4):410-418. doi: 10.1016/j.joen.2023.01.009. Epub 2023 Feb 8.
Prostaglandin E (PGE) 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 PGE-transporting pathway in human dental pulp and explore the relevance of PGE receptors (EP2/EP4) to angiogenesis and dentinogenesis.
Protein localization of microsomal PGE (mPGES)synthase, PGE transporters (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.
MRP4 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).
PGE-producing enzyme (mPGES), transporters (MRP4 and PGT), and PGE-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 PGE binding to EP2/EP4 is associated with angiogenic and dentinogenic responses.
前列腺素E（PGE）通过其包括细胞内合成、细胞外转运和受体结合的转运途径发挥生物学作用。本研究旨在确定PGE转运途径各组分在人牙髓中的定位，并探讨PGE受体（EP2/EP4）与血管生成和牙本质生成的相关性。
使用双重免疫荧光染色分析微粒体PGE（mPGES）合成酶、PGE转运体（多药耐药相关蛋白4 [MRP4]和前列腺素转运体[PGT]）以及EP2/EP4的蛋白定位。将人第三磨牙的牙片在有或无布他前列素（EP2激动剂）或利文前列素（EP4激动剂）的条件下培养1周。进行内皮细胞丝状伪足的形态计量分析以评估血管生成，并进行实时聚合酶链反应以评估血管生成和成牙本质细胞分化标志物。
MRP4和PGT在成牙本质细胞和内皮细胞中与mPGES以及EP2/EP4共定位。此外，MRP4在表达人白细胞抗原-DR的树突状细胞中与mPGES和EP4共定位。在牙片培养中，EP2/EP4激动剂诱导丝状伪足的数量和长度以及血管生成标志物（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（牙本质涎磷蛋白和I型胶原）的mRNA表达显著增加。
产生PGE的酶（mPGES）、转运体（MRP4和PGT）以及PGE特异性受体（EP2/EP4）在人牙髓的各种细胞成分中被免疫定位。EP2/EP4激动剂促进内皮细胞丝状伪足的生成并上调与血管生成和成牙本质细胞分化相关的基因，提示PGE与EP2/EP4的结合与血管生成和牙本质生成反应相关。
