口腔医学研究

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (12): 1153-1156.DOI: 10.13701/j.cnki.kqyxyj.2020.12.016

• 骨生物学研究 • 上一篇    下一篇

巨噬细胞极化在去势小鼠骨质疏松中的实验研究

罗涛, 郑志超, 黄江勇, 郭吕华, 吴哲, 张清彬*   

  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
  • 收稿日期:2020-03-04 出版日期:2020-12-28 发布日期:2020-12-28
  • 通讯作者: *张清彬,E-mail:doctorqingbin@hotmail.com
  • 作者简介:罗涛(1983~ ),男,湖北人,讲师,博士,主要从事口腔种植与修复相关工作与研究。
  • 基金资助:
    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)

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

LUO Tao, ZHENG Zhichao, HUANG Jiangyong, GUO Lvhua, WU Zhe, ZHANG Qingbin*   

  1. 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
  • Received:2020-03-04 Online:2020-12-28 Published:2020-12-28

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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。

关键词: 骨质疏松, 巨噬细胞, 细胞极化, 白细胞介素-10

Abstract: 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.

Key words: osteoporosis, macrophage, polarization, IL-10