IL-33刺激原代培养的小鼠骨髓来源肥大细胞脱颗粒并分泌IL-1β、IL-6及TNF-α  

IL-33 promotes degranulation of mouse bone marrow-derived mast cells and release of cytokines IL-1β,IL-6 and TNF-α

作　　者：周佳[1,2] 张晨[1] 尚靖[1] 

机构地区：[1]中国药科大学新药筛选中心,江苏南京210009 [2]江南大学药学院,江苏无锡214122

出　　处：《细胞与分子免疫学杂志》2016年第4期462-465,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：十二五国家支撑计划(2012BAI30B00)

摘　　要：目的检测白细胞介素33(IL-33)对小鼠骨髓来源肥大细胞(BMMC)脱颗粒及细胞因子分泌的影响。方法原代培养小鼠BMMC,用(0、10、20、50)ng/m L的IL-33进行刺激,分别收集细胞及培养上清。Western blot法检测c-Kit表达水平;于收集的各IL-23处理组培养上清中分别加入β己糖胺酶底物,通过底物与β己糖胺酶反应所得产物吸光度值,间接测定上清中β己糖胺酶含量。ELISA测定细胞上清液中组织胺含量评估肥大细胞脱颗粒情况;ELISA检测培养细胞上清液中IL-1β、IL-6及肿瘤坏死因子α(TNF-α)的水平。结果 IL-33促进BMMC的c-Kit表达。不同剂量的IL-33作用于BMMC 30 min后,可浓度依赖性地促进β己糖胺酶及组织胺的释放。IL-33作用于BMMC 24 h可促进IL-1β分泌,以50 ng/m L IL-33时作用最显著;IL-33可促进IL-6分泌,以50 ng/m L浓度时作用最显著;IL-33可促进TNF-α分泌,以20 ng/m L浓度时作用最显著。结论 IL-33可刺激BMMC脱颗粒,并分泌IL-1β、IL-6及TNF-α。Objective To investigate the effect of interleukin 33（ IL-33） on degranulation and cytokine release of mouse bone marrow-derived mast cells（ BMMCs）. Methods Mouse BMMCs were isolated and stimulated by 0,10,20,50 ng / m L IL-33. The expression of c-Kit was assessed by Western blotting. Beta-hexosaminidase content in culture supernatant was evaluated indirectly through the absorbance value of the product of the reaction between chromogenix substrate andβ-hexosaminidase. The levels of histamine and cytokines（ IL-1β,IL-6 and TNF-α） in culture supernatant were examined by ELISA. Results IL-33 induced the expression of c-Kit in BMMCs. Treatments with different concentrations of IL-33 for 30 minutes induced the degranulation of BMMCs to release β-hexosaminidase and histamine in a dose-dependent manner. IL-33 induced the release of IL-1β,IL-6 and TNF-α in BMMCs after treatments for 24 hours; the peak values of the three kinds of cytokines were got respectively in 50,50 and 20 ng / m L IL-33 treatment groups. Conclusion IL-33 could induce the degranulation of mast cells and the release of cytokines（ IL-1β,IL-6 and TNF-α）.

关 键 词：肥大细胞 白细胞介素33(IL-33) 脱颗粒 细胞因子 

分 类 号：R392.12[医药卫生—免疫学] R965[医药卫生—基础医学]