上海口腔医学

上海口腔医学 ›› 2025, Vol. 34 ›› Issue (1): 1-6.doi: 10.19439/j.sjos.2025.01.001

• 论著 • 上一篇    下一篇

牙龈素提取物对小鼠焦虑样行为的影响

赵旭日1, 江旖婷2, 张桓瑜2, 徐丽娜2, 何智妍1, 周薇1   

  1. 1.上海交通大学医学院附属第九人民医院 口腔微生态与系统性疾病实验室, 2.牙周病科, 上海交通大学口腔医学院, 国家口腔医学中心, 国家口腔疾病临床医学研究中心, 上海市口腔医学重点实验室, 上海市口腔医学研究所, 上海 200011
  • 收稿日期:2024-08-01 修回日期:2024-08-21 发布日期:2025-03-05
  • 通讯作者: 周薇,E-mail：sweetzw@hotmail.com
  • 作者简介:赵旭日（1995-）,男,硕士研究生,E-mail：gracexuri@outlook.com
  • 基金资助:
    国家自然科学基金（81971299）;上海市“科技创新行动计划”自然科学基金（22ZR1437500）;上海交通大学医学院附属第九人民医院交叉基金（JYJC202308）

Effect of gingipain extracts on anxiety-like behavior in mice

ZHAO Xu-ri1, JIANG Yi-ting2, ZHANG Huan-yu2, XU Li-na2, HE Zhi-yan1, ZHOU Wei1   

  1. 1. Laboratory of Oral Microbiota and Systemic Disease, 2.Department of Periodontology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology. Shanghai 200011, China
  • Received:2024-08-01 Revised:2024-08-21 Published:2025-03-05

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摘要： 目的: 探讨牙龈素提取物对小鼠焦虑样行为的影响。方法: 培养牙龈卟啉单胞菌（Porphyromonas gingivalis,P. gingivalis）ATCC33277菌株,采用超声结合超速离心的方法获得牙龈素（gingipains）提取物并利用免疫印迹法和酶活性实验鉴定。通过腹腔注射牙龈素提取物和（或）碘乙酰胺（Iodoacetamide,IAM）构建小鼠牙龈素感染模型。利用酶活性实验检测小鼠血浆中精氨酸特异性牙龈素（arginine-gingipain,Rgp）和赖氨酸特异性牙龈素（lysine-gingipain,Kgp）的活性。通过旷场实验和高架零迷宫实验检测小鼠的焦虑样行为。利用脑组织切片荧光染色观察脑内星形胶质细胞的变化。采用GraphPad Prism对数据进行统计学分析。结果: 培养牙龈卟啉单胞菌ATCC33277,得到具有Rgp和Kgp活性的牙龈素提取物。牙龈素提取物经腹腔注射后可在小鼠血浆中检测到Rgp和Kgp活性。与对照组相比,牙龈素组小鼠自主活动能力无显著变化,旷场实验中在中心区域活动时间显著减少（P<0.05）,高架零迷宫实验中进入开放区域频率显著减少（P<0.05）,焦虑样行为明显加强。免疫荧光染色显示,牙龈素组小鼠海马区域星形胶质细胞明显激活。以上结果均可被牙龈素抑制剂IAM明显缓解。结论: 牙龈素提取物可引起小鼠焦虑样行为,可能与星形胶质细胞活化导致的神经炎症有关。

关键词: 牙龈素, 焦虑样行为, 牙龈卟啉单胞菌, 旷场实验, 高架零迷宫实验

Abstract: PURPOSE: To observe the effect of gingipain extracts on anxiety-like behavior in mice. METHODS: Gingipain extracts were derived from Porphyromonas gingivalis(P. gingivalis) ATCC33277 by sonication and ultra-speed centrifugation and identified by Western blot and enzyme activity assay. Gingipains infection models were established by intraperitoneal injection of gingipain extracts with/without iodoacetamide (IAM). Enzyme activity assays were used to detect the activity of arginine-gingipain (Rgp) and lysine-gingipain (Kgp) in plasma of mice. Open field experiments and elevated zero maze experiments were used to detect anxiety-like behavior in mice. Immunofluorescence staining of brain tissue slices was used to observe changes in astrocytes of the brain. GraphPad Prism software was used for statistical analysis. RESULTS: The gingipain extracts were collected from P. gingivalis ATCC33277 and they showed Rgp and Kgp activity. The activity of Rgp and Kgp could be detected in mice plasma after intraperitoneal injection of gingipain extracts. Compared with the control group, there were no significant changes in the autonomous activity ability of the gingipain group mice. In the open field experiment, the central area staying time of the gingipain groups was significantly reduced(P<0.05). In the elevated zero maze experiment, the open area frequency of the gingipain groups was obviously shortened(P<0.05). Anxiety-like behavior was significantly enhanced. Immunofluorescence staining showed that gingipain extracts could significantly activate astrocytes in the hippocampus of mice. The above results could be significantly alleviated by gingipain inhibitor IAM. CONCLUSIONS: Gingipain extracts could induce anxiety-like behavior in mice, which may be related to neuroinflammation due to activation of astrocytes.

Key words: Gingipains, Anxiety-like behavior, Porphyromonas gingivalis, Open filed experiment, Elevated zero maze experiment

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