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

发布时间：2018-04-02 19:32

本文选题：肥大细胞　切入点：白细胞介素(IL-)　出处：《细胞与分子免疫学杂志》2016年04期

【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。
[Abstract]:Objective to detect interleukin 33 (IL-33) of mouse bone marrow-derived mast cell (BMMC) effect of degranulation and cytokine secretion. Methods primary cultured mouse BMMC, with ng/m L IL-33 (0,10,20,50) stimulation, the cells were collected and the supernatant of.Western blot were used to detect the expression level of c-Kit in the collection; all IL-23 treated cultured beta hexosaminidase substrates were added in the supernatant by substrate and beta hexosaminidase reaction products of absorbance, the indirect determination of evaluation of mast cells degranulation histamine content in the supernatant was determined.ELISA beta hexosaminidase content in the supernatant; ELISA detection of cell culture supernatant of IL-1 beta, and IL-6 tumor necrosis factor alpha (TNF- alpha) level. The results showed that IL-33 promotes BMMC expression of c-Kit. Different doses of IL-33 to BMMC 30 min after the release of.IL in a dose-dependent manner to promote beta hexosaminidase and amino groups The effect of -33 on BMMC 24 h can promote the secretion of IL-1 beta, with 50 ng/m L IL-33 is the most significant effect; IL-33 can promote the secretion of IL-6, with 50 ng/m L when the concentration of the most significant effect; IL-33 can promote TNF- secretion, with 20 ng/m concentration of L is the most significant. Conclusion IL-33 can stimulate the degranulation of BMMC, and the secretion of IL-1 beta, TNF- alpha and IL-6.

【作者单位】：中国药科大学新药筛选中心;江南大学药学院;
【基金】：十二五国家支撑计划(2012BAI30B00)
【分类号】：R758.41

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