Yu Yangyang, Lin Dongxu, Liu Zhenyu, Fang Ran, Zheng Siman, Cheng Yongxian, Huang Zhong, Ng Chun Wai, Lau Hang Yung Alaster
Shenzhen University Health Science Center, Shenzhen, China.
Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Iran J Basic Med Sci. 2022 May;25(5):629-634. doi: 10.22038/IJBMS.2022.64132.14120.
Mast cells are important immune cells that primarily localize in the interface between the host and external environment, and protect us from pathogen infection. However, they are also involved in the pathology of allergic diseases such as asthma and atopic dermatitis. A novel S phase kinase-associated protein 1 (SKP1) inhibitor 6-O-angeloylplenolin (6-OAP), was studied with its potential ability to alleviate the anti-IgE-induced inflammatory responses of primary human cultured mast cells (HCMCs) and LAD2 cell line.
We isolated the HCMCs from the buffy coat of voluntary blood donors. The effects of 6-OAP on mast cell activation were evaluated by measuring degranulation, cytokine release, migration, calcium influx, and ERK phosphorylation using spectro-fluorescence assay, multiplex cytometric bead assay/ELISA, migration assay, Fluo-4 calcium flux assay, and western blot, respectively.
It was found that 6-OAP exerted anti-inflammatory effects on human mast cells by dose-dependently suppressing the anti-IgE-mediated degranulation and release of cytokines such as proinflammatory cytokines (IL-8 and TNF-α), growth factors (GM-CSF, VEGF, and FGF), and chemokines (CCL2 and CCL3) in HCMC and LAD2 cells. It also suppressed the migration of immature HCMCs induced by CXCL12. Moreover, the process of calcium influx and ERK phosphorylation in activated HCMC cells were inhibited by 6-OAP administration.
Our results showed that 6-OAP inhibited anti-IgE-induced inflammatory responses of human mast cells via suppressing calcium influx and ERK phosphorylation.
肥大细胞是重要的免疫细胞，主要定位于宿主与外部环境的界面，保护我们免受病原体感染。然而，它们也参与哮喘和特应性皮炎等过敏性疾病的病理过程。研究了一种新型S期激酶相关蛋白1（SKP1）抑制剂6-O-当归酰蛇麻脂素（6-OAP）减轻抗IgE诱导的原代人培养肥大细胞（HCMC）和LAD2细胞系炎症反应的潜在能力。
我们从自愿献血者的血沉棕黄层中分离出HCMC。分别使用光谱荧光测定法、多重细胞计数珠测定法/酶联免疫吸附测定法、迁移测定法、Fluo-4钙流测定法和蛋白质免疫印迹法，通过测量脱颗粒、细胞因子释放、迁移、钙内流和ERK磷酸化来评估6-OAP对肥大细胞活化的影响。
发现6-OAP通过剂量依赖性抑制抗IgE介导的HCMC和LAD2细胞中的脱颗粒以及促炎细胞因子（IL-8和TNF-α）、生长因子（GM-CSF、VEGF和FGF）和趋化因子（CCL2和CCL3）等细胞因子的释放，对人肥大细胞发挥抗炎作用。它还抑制了CXCL12诱导的未成熟HCMC的迁移。此外，给予6-OAP可抑制活化的HCMC细胞中的钙内流和ERK磷酸化过程。
我们的结果表明，6-OAP通过抑制钙内流和ERK磷酸化来抑制抗IgE诱导的人肥大细胞炎症反应。
