口腔医学研究

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (1): 62-67.DOI: 10.13701/j.cnki.kqyxyj.2023.01.012

• 口腔黏膜病学研究 • 上一篇    下一篇

唾液链球菌对小鼠口腔黏膜慢性炎症的治疗作用

张诗, 徐盼, 郭艺婷, 邵如如, 赵若闻, 何园*   

  1. 上海牙组织修复与再生工程技术研究中心,同济大学口腔医学院,同济大学附属口腔医院口腔黏膜教研室 上海 200072
  • 收稿日期:2022-05-15 发布日期:2023-01-28
  • 通讯作者: *何园,E-mail:drheyuan@tongji.edu.cn
  • 作者简介:张诗(1995~ ),女,上海人,硕士,主要从事口腔黏膜疾病治疗相关研究。
  • 基金资助:
    国家自然科学基金(编号:82170961、81870764)

Therapeutic Effect of Streptococcus Salivarius on Chronic Inflammation of Oral Mucosa in Mice

ZHANG Shi, XU Pan, GUO Yiting, SHAO Ruru, ZHAO Ruowen, HE Yuan*   

  1. Department of Oral Medicine, Stomatological Hospital and Dental School of Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China
  • Received:2022-05-15 Published:2023-01-28

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摘要： 目的:利用口腔黏膜慢性炎症模型,对唾液链球菌(Streptococcus salivarius,Ss)的抗炎作用进行初步研究。方法:(1)健康成年SPF级C57BL/6小鼠75只,随机分为5组,空白对照(CON)组、慢性炎症(CI)组、唾液链球菌治疗(CI+SS)组、糖皮质激素治疗(CI+GC)组和单独使用唾液链球菌(CON+SS)组各15只。4周后收集病损部颊黏膜,经苏木素-伊红(HE)染色后观察其病理变化及炎性细胞浸润情况,采用实时荧光定量聚合酶链反应(qRT-PCR)方法检测病损局部白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、白细胞介素-17(IL-17)和叉头框蛋白3(Foxp3)的mRNA表达改变,采用免疫荧光染色法检测病损局部IL-6的蛋白表达情况。(2)健康成年SPF级C57BL/6小鼠45只,随机分为3组,CON组、CI组及CI+SS组各15只。4周后收集外周血,通过酶联免疫吸附法检测血清中IL-17A的表达水平,采用流式细胞术检测外周血Th17/Treg细胞比例。结果:(1)与CI组相比,CI+SS组病损局部炎性细胞浸润减少,IL-1β、IL-6和TNF-α的表达降低,IL-6阳性细胞减少(P＜0.001);(2)CI+SS组病损组织中IL-10、IL-17和Foxp3的表达降低(P＜0.01),血清IL-17A的表达降低(P＜0.001),外周血Th17/Treg细胞比例下降(P＜0.001)。结论:Ss局部使用具有改善口腔黏膜慢性炎症的作用,其机制可能是通过改善Th17/Treg分化失衡从而维持局部免疫平衡。

关键词: 唾液链球菌, 慢性炎症, 治疗

Abstract: Objective: To preliminarily study the anti-inflammatory effect of Streptococcus salivarius (Ss) by using a chronic inflammation model of oral cavity. Methods: Seventy-five C57BL/6 mice were randomly divided into 5 groups with 15 mice in each group. The first group was the blank control (CON) group; the second group was the chronic inflammation (CI) group; the third group was the Streptococcus salivarius treatment (CI+SS) group; the fourth group was the glucocorticoid treatment (CI+GC) group; and the fifth group was the Streptococcus salivarius alone (CON+SS) group. The mice were sacrificed after 4 weeks, and the buccal mucosa of the lesion was stained with HE to observe its pathological changes and inflammatory cell infiltration. The mRNA expressions of IL-1β, IL-6, TNF-α, IL-10, IL-17 and Foxp3 were detected by qRT-PCR, and the protein expression of IL-6 in the lesions was detected by immunofluorescence staining. Then, 45 C57BL/6 mice were randomly divided into 3 groups with 15 mice in each group. The first group was the CON group; the second group was the CI group; and the third group was the CI+SS group. After 4 weeks of treatment, the blood was collected. The expression levels of IL-17A in serum were detected by ELISA, and the ratio of Th17/Treg cells in peripheral blood was detected by flow cytometry. Results: Compared with the CI group, the CI+SS group had less local inflammatory cell infiltration, less expressions of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, and decreased IL-6 positive cells (P＜0.001). The expressions of IL-10, IL-17, and Foxp3 in the lesions of the CI+SS group were decreased (P＜0.01), the expression of serum IL-17A was decreased (P＜0.001), and the ratio of Th17/Treg cells in peripheral blood was decreased (P<0.001). Conclusion: Ss has a therapeutic effect on local inflammation of the oral mucosa, possibly by improving the imbalance of Th17/Treg differentiation to maintain local immune balance.

Key words: Streptococcus salivarius, chronic inflammation, treatment