Department of Pharmacology, Graduate School of Medicine, Ehime University, Toon, Ehime, 791-0295, Japan.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia.
Inflamm Res. 2020 Mar;69(3):289-298. doi: 10.1007/s00011-020-01319-z. Epub 2020 Jan 28.
Mice and rats are important animal models for mast cell (MC) study. However, rat Mas-related-GPCR-B3 receptor (MRGPRB3) has been less studied than its mouse counterpart. Therefore, we aimed to characterize rat MRGPRB3.
Mrgprb3 mRNA expression was assessed in peritoneal cells (RPCs) and peritoneal MCs (RPMCs) of wild-type rats, RPCs of MC-deficient rats, and RBL-2H3 cells by reverse-transcriptase polymerase chain reaction (RT-PCR). RPMCs, MRGPRX2-transfected and non-transfected RBL-2H3 cells were activated by 15-30 min incubation with DNP-BSA, substance-P (SP), or compound-48/80. L732138 or CP96344 was used as a tachykinin/neurokinin-1-receptor antagonist. Histamine release from MCs was measured by HPLC fluorometry.
Mrgprb3 mRNA expression was found in all cells, with the highest level in wild-type RPCs. All cells responded to DNP-BSA, but only MRGPRX2-transfected-RBL-2H3 cells and RPMCs responded to all activators. L732138 (0.1-10 μM) and CP96344 (1-100 μM) suppressed SP (10 μM)-induced RPMC activation. L732138 inhibition was dose independent, whereas CP96344 inhibition occurred in a dose-dependent manner. Additionally, only CP96344 suppressed SP (100 μM)- and compound-48/80 (10 μg/mL)-induced RPMC activation.
RPMCs expressing functional MRGPRB3 response upon MRGPRX2 ligands to regulated MC-mediated activities. It`s provide novel insights for future pseudo-allergic studies in rodents.
老鼠是研究肥大细胞（MC）的重要动物模型。然而，与小鼠对应的 Mas 相关 G 蛋白偶联受体 B3 受体（MRGPRB3）相比，其在大鼠中的研究较少。因此，我们旨在表征大鼠 MRGPRB3。
通过逆转录聚合酶链反应（RT-PCR）评估野生型大鼠腹腔细胞（RPCs）和腹腔 MC（RPMCs）、MC 缺陷型大鼠 RPCs 和 RBL-2H3 细胞中的 Mrgprb3 mRNA 表达。用 DNP-BSA、P 物质（SP）或化合物 48/80 孵育 15-30 分钟激活 RPMCs、MRGPRX2 转染和未转染的 RBL-2H3 细胞。L732138 或 CP96344 用作速激肽/神经激肽-1 受体拮抗剂。通过 HPLC 荧光法测量 MC 释放的组胺。
在所有细胞中均发现 Mrgprb3 mRNA 表达，在野生型 RPCs 中表达水平最高。所有细胞均对 DNP-BSA 有反应，但只有 MRGPRX2 转染的 RBL-2H3 细胞和 RPMCs 对所有激活剂有反应。L732138（0.1-10 μM）和 CP96344（1-100 μM）抑制 SP（10 μM）诱导的 RPMC 激活。L732138 的抑制作用呈剂量依赖性，而 CP96344 的抑制作用呈剂量依赖性。此外，只有 CP96344 抑制 SP（100 μM）和化合物 48/80（10 μg/mL）诱导的 RPMC 激活。
表达功能性 MRGPRB3 的 RPMCs 对 MRGPRX2 配体作出反应，调节 MC 介导的活性。这为未来啮齿动物的拟变应性研究提供了新的见解。
