| 摘要: |
| 目的 建立通过流式细胞分选术区分中枢神经系统内小胶质细胞和浸润巨噬细胞的方法。方法 用成年 C57BL/6 小鼠建立双侧颈总动脉狭窄模型,分别采用集落刺激因子 1 受体抑制剂 PLX5622 或氯膦酸盐脂质体处理。将分离、匀浆、重悬后的小鼠脑和脊髓组织进行 Percoll 密度梯度离心,得到单核细胞悬液。采用 CD45、CD11b 和淋巴细胞抗原 6 家族成员 C(Ly6C)抗体进行流式分选,获得小胶质细胞(CD11b+CD45lowLy6C-细胞)和浸润巨噬细胞(CD11b+CD45highLy6C+细胞),并验证 PLX5622 和氯膦酸盐脂质体 2 种给药范式获得的处理效果。结果 通过CD45、CD11b 和 Ly6C 抗体可以有效区分中枢神经系统中小胶质细胞和浸润巨噬细胞。与对照组比较, PLX5622 处理后小胶质细胞数量减少(P=0.001),而氯膦酸盐脂质体处理后浸润巨噬细胞数量减少(P<0.001)。结论 所建立的流式细胞分选方法可有效区分中枢神经系统中小胶质细胞和浸润巨噬细胞, PLX5622 和氯膦酸盐脂质体 2 种给药范式可分别选择性清除中枢神经系统内的小胶质细胞和浸润巨噬细胞。 |
| 关键词: 小胶质细胞 浸润巨噬细胞 流式细胞分选术 双侧颈总动脉狭窄 PLX5622 氯膦酸盐脂质体 |
| DOI：10.16781/j.CN31-2187/R.20230451 |
| 投稿时间：2023-08-08修订日期：2023-11-09 |
| 基金项目:军事医学基金(16QNP085),国家自然科学基金面上项目(82371357). |
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| Distinguishing microglia and infiltrating macrophages in central nervous system by fluorescence-activated cell sorting |
| ZHOU Jintao,YU Zhongwang*,CAO Li* |
(Department of Neurobiology, College of Basic Medical Sciences, Naval Medical University(Second Military Medical University), Shanghai 200433, China
*Corresponding authors) |
| Abstract: |
| Objective To establish a method for distinguishing microglia and infiltrating macrophages in central nervous system by fluorescence-activated cell sorting. Methods Adult C57BL/6 mice with bilateral common carotid artery stenosis were treated with PLX5622 (a colony stimulating factor 1 receptor inhibitor) or clodronate liposomes. After separating, homogenizing and resuspending brain and spinal cord tissue, the mononuclear cell suspensions were obtained by Percoll density gradient centrifugation. Anti-CD45, anti-CD11b, and anti-lymphocyte antigen 6 family member C (Ly6C) were used to obtain microglia (CD11b+CD45lowLy6C- cells) and infiltrating macrophages (CD11b+CD45highLy6C+ cells) by fluorescence-activated cell sorting. The treatment effects of PLX5622 and clodronate liposomes were verified. Results Microglia and infiltrating macrophages in the central nervous system were effectively distinguished by anti-CD45, anti-CD11b, and anti-Ly6C. Compared with the control group, the number of microglia was significantly decreased after PLX5622 treatment (P=0.001), while the number of infiltrating macrophages was significantly decreased after clodronate liposome treatment (P<0.001). Conclusion The established method can effectively distinguish microglia and infiltrating macrophages in central nervous system by fluorescence-activated cell sorting, and PLX5622 and clodronate liposomes can selectively eliminate microglia and infiltrating macrophages in the central nervous system. |
| Key words: microglia infiltrating macrophage fluorescence-activated cell sorting bilateral common carotid artery stenosis PLX5622 clodronate liposomes |
