口腔医学研究 ›› 2021, Vol. 37 ›› Issue (5): 412-417.DOI: 10.13701/j.cnki.kqyxyj.2021.05.008

• 龋病牙髓病学研究 • 上一篇    下一篇

巨噬细胞极化在糖尿病合并根尖周炎中作用的研究

孙熹1,2, 张旗1,2*   

  1. 1.同济大学口腔医学院·同济大学附属口腔医院牙体牙髓科 上海 200072;
    2.上海牙组织修复与再生工程技术研究中心 上海 200072
  • 收稿日期:2021-02-07 发布日期:2021-05-17
  • 通讯作者: * 张旗,E-mail:qizhang@tongji.edu.cn
  • 作者简介:孙熹(1994~ ),女,上海人,硕士在读,研究方向:伴糖尿病患者牙髓根尖周病诊疗。

Effect of Macrophage Polarization in Apical Periodontitis Complicated with Diabetes Mellitus

SUN Xi1,2, ZHANG Qi1,2*   

  1. 1. Department of Endodontics, School and Hospital of Stomatology, Tongji University, Shanghai 200072, China;
    2. Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China
  • Received:2021-02-07 Published:2021-05-17

摘要: 目的:探究巨噬细胞极化在糖尿病合并根尖周炎中作用,及高糖环境对巨噬细胞极化影响。方法:8周龄雄性Wistar大鼠(对照组)和GK大鼠(糖尿病组)构建根尖周炎模型,2周、4周取材。苏木素-伊红(hematoxylin-eosin,HE)染色观察根尖炎症浸润;免疫组化观察炎症因子表达;免疫荧光观察巨噬细胞极化情况。体外培养Raw264.7小鼠巨噬细胞系,高浓度葡萄糖(40 mmol/L)处理,实时荧光定量聚合酶链反应(quantitative real-time PCR)检测巨噬细胞极化及炎症因子相关mRNA表达水平。结果:建模4周,与对照组相比,糖尿病组根尖炎症浸润增多,促炎因子TNF-α、IL-1β、IL-6增多,抗炎因子IL-10减少(P<0.05);M2型巨噬细胞占总巨噬细胞比值(CD206/CD68)减少,巨噬细胞iNOS表达增多,凋亡细胞数目增多(P<0.05)。高糖处理Raw264.7小鼠巨噬细胞系,iNOS mRNA表达升高,CD206表达降低,CD68表达无明显差异;TNF-α、IL-1β、IL-6 mRNA 表达升高,IL-10 mRNA表达降低(P<0.05)。结论:糖尿病所致高糖环境通过影响巨噬细胞极化,促进炎性浸润,加重根尖周炎。

关键词: 根尖周炎, 糖尿病, 巨噬细胞极化

Abstract: Objective: To explore the role of macrophage polarization in apical periodontitis complicated with diabetes mellitus and the effect of high glucose environment on macrophage polarization. Methods: Eight-week-old male Wistar rats (control group) and GK rats (diabetes group) were used to construct the apical periodontitis model, and samples were taken at 2 weeks and 4 weeks. Hematoxylin-eosin (HE) staining was used to observe the infiltration of periapical inflammation. The expression of inflammatory cytokines was visualized by immunohistochemical staining. Immunofluorescence staining was used to evaluate the polarization of macrophages. Raw264.7 mouse macrophage lines were cultured in vitro and treated with high concentration of glucose (40 mM). Quantitative real-time PCR was used to detect the mRNA expression of iNOS, CD206, CD68, IL-1β, IL-6, TNF-α, and IL-10. Results: After 4 weeks, apical inflammatory infiltration in the diabetic group was significantly higher than that in the control group. Immunohistochemical staining showed that the expression of pro-inflammatory cytokines TNF-α, IL-1β and IL-6 increased in the diabetic group, while the expression of anti-inflammatory cytokines IL-10 decreased in the diabetic group (P<0.05). The ratio of M2 macrophages to total macrophages in the diabetic group (CD206/CD68) was lower than that in the control group, and the expression of iNOS in macrophages as well as the number of apoptotic cells were both increased (P<0.05). In vitro, the mRNA expression of iNOS increased while CD206 decreased, and there was no significant difference in the mRNA expression of CD68. The level of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 ascended, and anti-inflammatory cytokines IL-10 descended (P<0.05). Conclusion: Diabetes-induced hyperglycemia can affect macrophage polarization and inflammatory cytokines secretion, which may further aggravate periapical periodontitis

Key words: apical periodontitis, diabetes, macrophage polarization