Department of Endocrinology and Metabolism, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
The Core Laboratory in Medical Center of Clinical Research, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Cell Physiol. 2018 Aug;233(8):6117-6124. doi: 10.1002/jcp.26455. Epub 2018 Mar 7.
The airway epithelium plays a crucial role in the pathogenesis of asthma. The functions of leukotriene B4 receptor 2 (BLT2) on the airway epithelial cells remains unknown. In our study, BLT2 expression in 16HBE bronchial epithelial cells were manipulated by transfection with BLT2 overexpression plasmid or BLT2 small interference RNA. 16HBE cells were then exposed to BLT2 antagonist (LY255283) or BLT2 agonist (12(S)-hydroxyheptadeca-5Z,8E,10E-trienoic acid [12-HHT] or CAY10583). The results showed that BLT2 overexpression, 12-HHT stimulation, or CAY10583 treatment resulted in the enhanced proliferation and migration of 16HBE cells. In addition, BLT2 showed an inhibitory effect on epithelial permeability as illustrated by the measurement of transepithelial electrical resistance (TER) and epithelial permeability, and a promoting effect on the levels of tight junction proteins (occludin and claudin-4) and phosphorylated p38 as demonstrated by real-time PCR and Western blotting analyses. These results suggest BLT2 as a key determinant of airway epithelial barrier integrity. On the contrary, RNAi-mediated knockdown or LY255283 treatment had reversed effects on the proliferation, migration, and epithelial barrier integrity. Together, our findings suggest the critical roles of BLT2 on the functions of bronchial epithelial cells and that BLT2 agonists are potential therapeutic agents for asthma treatment.
气道上皮在哮喘发病机制中起着至关重要的作用。白细胞三烯 B4 受体 2 (BLT2) 在气道上皮细胞上的功能尚不清楚。在我们的研究中,通过转染 BLT2 过表达质粒或 BLT2 小干扰 RNA 来操纵 16HBE 支气管上皮细胞中的 BLT2 表达。然后将 16HBE 细胞暴露于 BLT2 拮抗剂 (LY255283) 或 BLT2 激动剂 (12(S)-羟基十七碳-5Z,8E,10E-三烯酸[12-HHT]或 CAY10583)。结果表明,BLT2 过表达、12-HHT 刺激或 CAY10583 处理可增强 16HBE 细胞的增殖和迁移。此外,BLT2 对上皮通透性具有抑制作用,表现为跨上皮电阻 (TER) 和上皮通透性的测量,以及对紧密连接蛋白 (occludin 和 claudin-4) 和磷酸化 p38 的水平具有促进作用,如实时 PCR 和 Western blot 分析所示。这些结果表明 BLT2 是气道上皮屏障完整性的关键决定因素。相反,RNAi 介导的敲低或 LY255283 处理对增殖、迁移和上皮屏障完整性具有逆转作用。总之,我们的研究结果表明 BLT2 在支气管上皮细胞功能中的关键作用,并且 BLT2 激动剂可能是哮喘治疗的潜在治疗剂。