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山东大学学报 (医学版) ›› 2021, Vol. 59 ›› Issue (3): 35-40.doi: 10.6040/j.issn.1671-7554.0.2021.0176

• 基础医学 • 上一篇    下一篇

牙周膜干细胞调节巨噬细胞功能的体外研究

张栌丹1,丁晓玲2,崔舒悦1,程晨1,魏福兰3,丁刚1   

  1. 1. 潍坊医学院口腔医学院, 山东 潍坊 261053;2. 潍坊医学院临床能力培训中心, 山东 潍坊 261053;3.山东大学口腔医学院, 山东 济南 250012
  • 发布日期:2021-04-06
  • 通讯作者: 丁刚. E-mail:dinggang@wfmc.edu.cn
  • 基金资助:
    国家自然科学基金(81570945);全牙再生与口腔组织功能重建北京市重点实验室开放课题(KFKT2019013)

Periodontal ligament stem cells regulate the functions of macrophages in vitro

ZHANG Ludan1, DING Xiaoling2, CUI Shuyue1, CHENG Chen1, WEI Fulan3, DING Gang1   

  1. 1. School of Stomatology, Weifang Medical University, Weifang 261053, Shandong, China;
    2. Clinical Competency Training Center, Weifang Medical University, Weifang 261053, Shandong, China;
    3. School of Stomatology, Shandong University, Jinan 250012, Shandong, China
  • Published:2021-04-06

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摘要： 目的 探讨牙周膜干细胞(PDLSCs)对外周血来源巨噬细胞的表型和功能的体外影响。 方法 分离、培养PDLSCs,并检测其间充质干细胞标志物基质细胞抗原-1(STRO-1)、表面抗原146(CD146)、表面抗原90(CD90)的表达情况及骨向、脂肪向分化能力。分离外周血来源的巨噬细胞。将PDLSCs与等量异体巨噬细胞在Transwell培养系统中37 ℃、5% CO2条件下共培养,为实验组。巨噬细胞的单独培养设置为对照组。共培养3 d后,提取巨噬细胞,采用流式细胞术检测CD14+CD206+巨噬细胞的表达情况;共培养24 h后,提取巨噬细胞,加入荧光素异硫氰酸酯标记的葡聚糖,孵育 30 min后,采用流式细胞术检测巨噬细胞的吞噬率;共同培养3 d后,取细胞培养上清,采用酶联免疫吸附法检测上清中白介素-10(IL-10)、白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的浓度。 结果 PDLSCs呈梭状的成纤维细胞样,表达STRO-1、CD146和CD90,可以骨向和脂肪向分化。与对照组相比:(1)共培养组的CD14+CD206+巨噬细胞表达率明显升高［(38.73±6.32)% vs(8.39±2.71)%,t=127.7,P=0.004 9］;(2)共培养组的巨噬细胞吞噬率无显著变化［(36.7±5.1)% vs(38.6±4.3)%,t=3.904,P=0.159 6］;(3)共培养组的IL-10浓度明显升高［(382.5±18.2)pg/mL vs (198.5±11.4)pg/mL, t=76.36,P=0.000 3］, IL-6浓度显著降低［(453.1±70.42)pg/mL vs(936.7±49.9)pg/mL, t=53.12,P=0.011 5］, TNF-α浓度也显著降低［(64.9±11.3)pg/mL vs(131.7±19.3)pg/mL, t=51.48,P=0.000 6］。 结论 PDLSCs可以促使巨噬细胞向M2型极化,不影响巨噬细胞的吞噬功能,促进抗炎因子IL-10的分泌,抑制炎性因子IL-6和 TNF-α的分泌。

关键词: 牙周膜干细胞, 巨噬细胞, 极化, Transwell共培养

Abstract: Objective To explore the effects of periodontal ligament stem cells(PDLSCs)on the phenotypes and functions of macrophages. Methods After PDLSCs were isolated and cultured, the expression profiles of STRO-1, CD146 and CD90, as well as the multipotent differentiation capabilities were detected. After macrophages were isolated from peripheral blood, they were cocultured with an equal amount PDLSCs in Transwell co-culture condition at 37 ℃ and 5% CO2, which were set as the experimental group. Macrophages cultured alone were set as the control group. After 3d co-culture, the expression profiles of CD14+CD206+ macrophages were examined by flow cytometry. After 24 h co-culture, macrophages were obtained, fluorescein isothiocyanate labeled dextran was added. Then, after 30 min incubation, the phagocytosis rate of macrophages was detected with flow cytometry. After 3 d co-culture, the supernatant was collected, and the concentrations of IL-10, IL-6 and TNF-α were determined with enzyme-linked immunosorbent assays. Results PDLSCs displayed fusiform fibroblast-like morphology, positive for the mesenchymal stem cells surface markers including STRO-1, CD146 and CD90, and could differentiate into bone cells and lipid cells. Compared with the control group, the experimental group had significantly increased expression of CD14+CD206+macrophages ［(38.73±6.32)% vs(8.39±2.71)%, t=127.7, P=0.004 9), unchanged phagocytosis rate of macrophages ［(36.7±5.1)% vs(38.6±4.3)%, t=3.904, P=0.159 6］, elevated level of IL-10 ［(382.5±18.2)pg/mL vs(198.5±11.4)pg/mL, t=76.36, P=0.000 3］, but decreased levels of IL-6 ［(453.1±70.42)pg/mL vs(936.7±49.9)pg/mL, t=53.12, P=0.011 5)and TNF-α ［(64.9±11.3)pg/mL vs(131.7±19.3)pg/mL, t=51.48, P=0.000 6］. Conclusion PDLSCs are capable of converting macrophages into M2 phenotype without affecting the phagocytic functions. Meanwhile, they can stimulate the secretion of IL-10 but inhibit the secretion of IL-6 and TNF-α.

Key words: Periodontal ligament stem cells, Macrophages, Polarization, Transwell co-culture system

中图分类号: 

  • R781
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