小鼠骨髓巨噬细胞雌激素膜表面受体的检测
Detection of estradiol membrane receptor on mouse bone marrow derivedmacrophage
刘立民 1谢可鸣 1孙晓东 1王祖峰2
作者信息
- 1. 苏州大学基础医学与生物科学学院病理学与病理生理学系,江苏苏州215123
- 2. 苏州大学基础医学与生物科学学院法医学系,江苏苏州215123
- 折叠
摘要
目的 使用不同方法检测小鼠骨髓巨噬细胞(BMM)雌激素膜受体,突破雌激素作用机制的传统观念,为临床更好地防治相关疾病提供新思路.方法 六周龄C57B L/6j雄性小鼠,体外诱导培养BMM.收集细胞,与17β-estradiol-BSA-FITC或BSA-FITC孵育,用激光共聚焦扫描显微镜和流式细胞仪检测其与巨噬细胞的结合状况.将巨噬细胞与17β-estradiol- BSA-FITC孵育的同时,分别加入十倍浓度的17β-estradiol、17β-estradiol-BSA、Tamoxifen、Testosterone,用流式细胞仪检测不同处理组荧光强度的变化.结果 激光共聚焦扫描显微镜显示17 β-estradiol-BSA-FITC结合于巨噬细胞膜上,而BSA-FITC对照组未检测到相应荧光.流式细胞仪检测显示17β-estradiol、17β-estradiol-BSA竞争性抑制17β-estradiol-BSA-FITC与巨噬细胞的结合,而Tamoxifen、Testosterone对17β-estradiol-BSA-FITC与巨噬细胞的结合无影响.结论 小鼠骨髓巨噬细胞表面存在雌激素膜受体.
Abstract
Objective Our project aims to provide reliable evidences that there are estradiol receptors on the membrane of murine bone marrow-derived macrophages ( BMMs) , which will break through the traditional idea of estadiol action and provide a novel strategy for healing some estradiol-related diseases in clinic. Methods BMMs were obtained from 6-week-old male C57BL/6J mice. BMMs were labeled with 17β-estradiol-BSA-FITC or BSA-FITC, and fluorescence binding was detected by confocal laser scanning microscopy and flow cytometry. Then, BMMs were incubated with 17β-estradiol-BSA-FITC in the presence of 10-fold excess of 17β-estradiol, 17p-estradiol-BSA, tamoxifen or testosterone, and fluorescence intensity was analyzed by flow cytometry. Results Confocal laser scanning microscopy showed that 17β-estradiol-BSA-FITC was able to bind on the plasma membrane of BMMs. Flow cytometry indicated that the binding of 17β-estradiol-BSA-FITC was competitively reduced by 17β-estradiol and 17|3-es-tradiol-BSA, whereas tamoxifen and testosterone had no influence on 17p-estradiol-BSA-FITC binding.Conclusion It is reasonable to assume that there are estradiol receptors on the surface of BMMs.
关键词
巨噬细胞/雌激素/膜受体Key words
macrophage/estradiol/membrane receptor引用本文复制引用
基金项目
国家自然科学基金(31000630)
江苏省自然科学基金资助项目(BK2010217)
出版年
2012
NETL
NSTL
维普
万方数据