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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C57BL/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17β-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。
Objective To detect the estrogen receptor of mouse bone marrow macrophage (BMM) by different methods and to break through the traditional conception of the mechanism of action of estrogen, so as to provide new ideas for better prevention and treatment of related diseases in clinic. Methods Six-week-old C57BL / 6j male mice were cultured in vitro to induce BMM. Cells were harvested, incubated with 17β-estradiol-BSA-FITC or BSA-FITC, and their binding to macrophages was examined by laser confocal scanning microscopy and flow cytometry. When the macrophages were incubated with 17β-estradiol-BSA-FITC, the concentrations of 17β-estradiol, 17β-estradiol-BSA, Tamoxifen and Testosterone were respectively added. The changes of fluorescence intensity of different treatment groups were detected by flow cytometry. Results Confocal laser scanning microscopy showed that 17β-estradiol-BSA-FITC bound to the macrophage membrane, while no fluorescence was detected in the BSA-FITC control group. Flow cytometry showed 17β-estradiol and 17β-estradiol-BSA competitively inhibited the binding of 17β-estradiol-BSA-FITC to macrophages, whereas Tamoxifen and Testosterone inhibited the binding of 17β-estradiol-BSA-FITC to macrophages no effect. Conclusion There is estrogen receptor on the surface of mouse bone marrow macrophage.