巨噬细胞极化在去势小鼠骨质疏松中的实验研究  

Effect of Ovariectomy on Macrophage Polarization in Bone Marrow:A Pilot Study in Mice

作　　者：罗涛[1] 郑志超 黄江勇[1] 郭吕华[1] 吴哲[1] 张清彬[1] LUO Tao;ZHENG Zhichao;HUANG Jiangyong;GUO Lvhua;WU Zhe;ZHANG Qingbin(Department of Prosthodontics, Affiliated Stomatology Hospital of Guangzhou Medical University, Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou 510182, China)

机构地区：[1]广州医科大学附属口腔医院修复科,广州市口腔再生医学基础与应用研究重点实验室,广东广州510182

出　　处：《口腔医学研究》2020年第12期1153-1156,共4页Journal of Oral Science Research

基　　金：国家自然科学基金青年科学基金项目(编号:81300902);广东省医学科学技术研究基金(编号:B2018099)。

摘　　要：目的:建立去势小鼠骨质疏松模型,探讨雌激素不足对巨噬细胞极化的调控作用。方法:选取8周龄健康雌性小鼠C57BL/6,随机分为手术去势组、对照组,每日测量体重,4周后取材,通过骨矿物质密度(bone mineralization density,BMD)测定确定骨质疏松模型的建立成功,心脏穿刺获取外周血,ELISA法检测小鼠的血清白细胞介素-10(interleukin-10,IL-10)水平,取股骨骨髓细胞,PCR检测骨髓细胞IL-10基因水平,流式细胞测定检测巨噬细胞M1/M2在手术去势组、对照组差异;体外培养巨噬细胞系细胞,检测IL-10对M1、M2相关基因的表达。结果:骨密度测定证实,去势组小鼠股骨密度低于对照组,血清ELISA检测发现,去势小鼠血清IL-10水平及骨髓细胞IL-10 mRNA基因表达明显降低(P<0.01),流式细胞检测提示去势小鼠骨髓巨噬细胞M1/M2比值较对照组上升(P<0.01),巨噬细胞向M1巨噬细胞型分化;机制上,IL-10处理促使巨噬细胞M2相关基因高表达。结论:雌激素分泌不足降低血清IL-10蛋白及骨髓细胞IL-10 mRNA表达,促进巨噬细胞向M1表型分化。Objective:To investigate the role of estrogen in regulating macrophage polarization by establishing a model of osteoporosis in ovariectomized mice.Methods:8-week-old healthy female C57BL/6 mice were randomly divided into ovariectomy group and control(pseudo-surgical)group.Body weights were measured every 3 days and animals were euthanized at 12-week-old.Bone mineral density was measured using DXA.Serum interleukin-10 concentration was determined by ELISA.Bone marrow cells were obtained from femoral bone and macrophages polarization were determined using flow cytometry.The ratio of M1/M2 in the ovariectomy group was compared to that of control group.Macrophage cells were cultured in vitro in the presence of IL-10 and the expression of M1/M2 related genes was detected using realtime-PCR.Results:Bone density measurement confirmed that ovariectomy induced bone loss in the femur(P<0.05).The ovariectomy group mice gained weight for 18 days.Ovariectomy group show significantly reduced serum IL-10(P<0.01)and bone marrow cells IL-10 mRNA expression(P<0.01),and increased M1 to M2 macrophages ratio(P<0.01).Mechanically,in vitro macrophage cell culture experiment confirmed that IL-10 induced upregulation of M2 macrophage-related genes(P<0.01).Conclusion:Insufficient estrogen secretion lowers serum IL-10 expression and bone marrow cells IL-10 mRNA expression,which enables macrophage differentiation toward M1 type.

关 键 词：骨质疏松 巨噬细胞 细胞极化 白细胞介素-10 

分 类 号：R73[医药卫生—肿瘤]