抑制ABCC1/MRP1转运体下调IL-1β分泌的实验研究
Study on Down-regulation of Interleukin-1β Secretion by Inhibiting ABCC1/MRP1 Transporter
陈园园 1应培挺 1翁雯雯 1方美新 1李江 1骆泽斌 1贾明 1郭晓萍 1张玲燕 1徐晓军 1汤永民1
作者信息
- 1. 浙江大学医学院附属儿童医院血液科,浙江杭州 310003
- 折叠
摘要
目的:用体外巨噬细胞实验和全基因敲除小鼠,筛选11.-1β分泌相关的胞膜转运体.方法:分化THP-1细胞株得到人源性巨噬细胞,从人外周血获取巨噬细胞.获取同性别、同年龄的FVB野生型小鼠作为MRP1全基因敲除小鼠的对照,培养小鼠腹腔和骨髓巨噬细胞.使用ABCC1/MRP1、ABCG2/BCRP、ABCB1/P-gp、OATP1B1和MATE转运体抑制剂处理细胞,再使用脂多糖和腺苷三磷酸刺激细胞,采用ELISA、Western blot和细胞免疫荧光法检测IL-1β分泌水平.结果:人和小鼠巨噬细胞抑制剂实验表明,抑制ABCC1/MRP1转运体后,IL-1β分泌显著降低,而抑制其他4类转运体无效果.在动物实验中,MRP1全基因敲除小鼠的骨髓巨噬细胞分泌IL-1β的水平显著低于对照组.结论:ABCC1/MRP1转运体是一种新发现的IL-1β分泌途径,有望成为解决细胞因子释放综合征等临床问题的新靶标.
Abstract
Objective:To screen interleukin(IL)-1β secretion-related membrane transporters by macrophage experiment in vitro and conventional knockout mice.Methods:THP-1 cell line was differentiated to obtain human THP-1-derived macrophages,and the primary macrophages were obtained from human peripheral blood.FVB wild-type mice with the same sex and age were used as the controls of MRP1 knockout mice.The macrophages in abdominal cavity and bone marrow of mice were cultivated.The cells were treated with ABCC1/MRP1,ABCG2/BCRP,ABCB1/P-gp,OATP1B1,and MATE transporter inhibitors,then stimulated by lipopolysaccharide and adenosine triphosphate.The secretion level of IL-iβ was detected by ELISA,Western blot,and immunofluorescence.Results:After inhibiting ABCC1/MRP1 transporter,the secretion of IL-1β decreased significantly,while inhibition of the other 4 transporters had no effect.In animal experiment,the level of IL-1 β secreted by macrophages in bone marrow of MRP1 knockout mice was significantly lower than control group(P＜0.05).Conclusion:ABCC1/MRP1 transporter is a newly discovered IL-1β secretion pathway,which is expected to become a new target for solving clinical problems such as cytokine release syndrome.
关键词
ABCC1/MRP1/巨噬细胞/白介素-1β/细胞因子释放综合征Key words
ABCC1/MRP1/macrophage/interleukin-1 β/cytokine release syndrome引用本文复制引用
基金项目
国家自然科学基金(81770202)
浙江省医药卫生科技计划(2022497122)
浙江省科技厅重点研发项目(2019C03032)
浙江省小儿白血病诊治技术研究中心项目(JBZX-201904)
出版年
2024
NETL
NSTL
维普
万方数据