苦精促进MrgprB2介导的鼠肥大细胞脱颗粒  

Denatonium benzoate promotes MrgprB 2-mediated rat mast cell degranulation

作　　者：徐华平[1] 石小云[1] 邹节新 李欣 谢梦婷 肖诗宇 石林波 XU Huaping;SHI Xiaoyun;ZOU Jiexin;LI Xin;XIE Mengting;XIAO Shiyu;SHI Linbo(The First Affiliated Hospital of Nanchang University,Nanchang 330006,China;School of Basic Medical Sciences,Jiangxi Medical College,Nanchang University,Nanchang 330006,China;School of Food Science and Technology,Nanchang University,Nanchang 330006,China;The First Clinical Medical School,Jiangxi Medical College,Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一附属医院,南昌330006 [2]南昌大学医学部基础医学院,南昌330046 [3]南昌大学食品学院,南昌330006 [4]南昌大学医学部第一临床医学院,南昌330006

出　　处：《中国免疫学杂志》2024年第10期2037-2041,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(21866020,32060306);江西省自然科学基金(20202BABL206108)。

摘　　要：目的:探讨微污染物苯甲地那铵(DB)对鼠Mas相关G蛋白偶联受体B2(MrgprB2)介导的肥大细胞(MCs)脱颗粒的影响,以较全面评估DB与以MCs脱颗粒为基础的过敏性疾病之间的关系。方法:建立MrgprB2介导的体外大鼠嗜碱性细胞白血病细胞株(RBL-2H3)活化模型,用不同剂量DB与RBL-2H3细胞共同孵育过夜,加入MrgprB2配体P物质(SP)按不同检测目的刺激RBL-2H3细胞不同时间后收集细胞或上清液,底物法检测MCs颗粒物质如β-氨基己糖苷酶(β-hex),ELISA检测白三烯C4(LTC4)、IL-6、TNF-α及细胞质磷脂酶A2(cPLA2)活性,荧光法检测MCs Ca^(2+)内流与细胞MrgprB2受体表达的变化。结果:10μmol/L、50μmol/L、80μmol/L、100μmol/L DB能促进MrgprB2介导的RBL-2H3细胞活化时β-hex、LTC4释放,且这一过程伴有细胞内Ca^(2+)内流的增加,且其效应呈一定的剂量依赖关系。此外,DB预处理对RBL-2H3内TNF-α和IL-6释放无影响,且不影响RBL-2H3上MrgprB2的表达增加。结论:DB促进MrgprB2介导的RBL-2H3细胞活化脱颗粒,其机制可能与通过促进MrgprB2中通路Ca^(2+)信号活化有关。Objective:To explore the potent effects of denatonium benzoate(DB)on Mas-related G protein-coupled receptor-B2(MrgprB2)-mediated rat mast cell degranulation.Methods:RBL-2H3 cells were treated with DB overnight,before challenged with MrgprB2 ligands substance P(SP).The release ofβ-hex from MrgprB2-activated RBL-2H3 was detected by substrate method.Detec⁃tion of LTC4,IL-6,TNF-αand cPLA2 activity were performed by ELISA.The Ca^(2+)influx and the expression of RBL-2H3 MrgprB2 re⁃ceptors were measured by fluorescence assay.Results:The results showed 10μmol/L,50μmol/L,80μmol/L,100μmol/L DB treat⁃ments promotedβ-hex and LTC4 releases from activated RBL-2H3,accompanied by increased Ca^(2+)mobilization and cPLA2 activa⁃tion.DB treatments did not affect IL-6 and TNF-αLTC4 releases in MrgprB2-activated RBL-2H3,as well as the levels of MrgprB2 ex⁃pression in mast cells.Conclusion:Taken together,DB enhanced the MrgprB2-mediated RBL-2H3 degranulation in vitro,probably by up-regulating Ca^(2+)mobilization in activated cells.

关 键 词：苯甲地那铵 MrgprB2 肥大细胞脱颗粒 Ca^(2+)活化 

分 类 号：R392.8[医药卫生—免疫学]