产黑普雷沃氏菌加重小鼠的牙周炎症并损伤认知功能

doi:10.12122/j.issn.1673-4254.2025.03.02

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (3): 453-460.doi: 10.12122/j.issn.1673-4254.2025.03.02

• • 上一篇

产黑普雷沃氏菌加重小鼠的牙周炎症并损伤认知功能

陈琦¹^,²(), 夏天天³^,⁴(), 周永强⁴, 常铭洋⁵, 胡楠², 杨燕美², 李仲², 高月⁴(), 顾斌²()

^1.解放军医学院，北京 100853
^2.解放军总医院第一医学中心口腔科，北京 100853
^3.青海大学医学院，西宁 810016
^4.中国人民解放军军事医学研究院，辐射医学研究所抗辐射药物研究室，北京 100850
^5.天津中医药大学，天津 300193

收稿日期:2024-11-27 出版日期:2025-03-20 发布日期:2025-03-28
通讯作者: 高月,顾斌 E-mail:CC313nn@163.com;xiatianttx@163.com;gaoyue@ bmi.ac.cn;gubinmail301@163.com
作者简介:陈琦，在读硕士研究生，医师，E-mail: CC313nn@163.com
夏天天，在读硕士研究生，E-mail: xiatianttx@163.com
第一联系人：陈琦、夏天天共同为第一作者
基金资助:
军队医学科技青年培育计划孵化项目(20QNPY100)

Prevotella nigrescens exacerbates periodontal inflammation and impairs cognitive function in mice

Qi CHEN¹^,²(), Tiantian XIA³^,⁴(), Yongqiang ZHOU⁴, Mingyang CHANG⁵, Nan HU², Yanmei YANG², Zhong LI², Yue GAO⁴(), Bin GU²()

^1.Chinese PLA Medical School, Beijing 100853, China
^2.Department of Stomatology, First Medical Center, Chinese PLA General Hospital, Beijing 100853, China
^3.Medical College of Qinghai University, Xining 810016, China
^4.Academy of Military Medical Sciences, Anti-radiation Drug Research Laboratory of Institute of Radiation Medicine, Beijing 100850, China
^5.Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China

Received:2024-11-27 Online:2025-03-20 Published:2025-03-28
Contact: Yue GAO, Bin GU E-mail:CC313nn@163.com;xiatianttx@163.com;gaoyue@ bmi.ac.cn;gubinmail301@163.com

摘要/Abstract

摘要：

目的探究产黑普雷沃氏菌（Pn）联合结扎诱导的牙周炎对小鼠认知功能的影响。方法将24只C57BL/6J小鼠随机分为3组：空白对照组（CON）、结扎组（P）、结扎+菌组（P+Pn）。P组丝线结扎诱导实验性牙周炎，P+Pn组丝线结扎并涂布产黑普雷沃氏菌（Pn）诱导实验性牙周炎。结扎、涂菌6周后，通过Micro CT、组织学观察牙槽骨吸收；通过旷场测试、新物体识别测试、Morris水迷宫测试评价其学习记忆能力改变；组织学观察小鼠大脑海马组织学变化。评价产黑普雷沃氏菌（Pn）致牙周炎及对小鼠认知功能的影响。结果牙周结扎6周，Micro CT显示小鼠牙槽骨水平吸收、根分叉暴露，组织学观察见小鼠牙周结合上皮根方退缩，上皮钉突增生、淋巴细胞浸润，P+Pn较P组变化更明显，牙槽骨高度均较CON组降低（P<0.05）。旷场测试的运动总路程、中央区时间占比、路程占比及新物体识别测试的路程占比、时间占比、次数占比均较CON组下降（P<0.05）；Morris水迷宫测试各项指标均下降（P<0.05），且P+Pn较P组下降更为明显（P<0.05）；组织学见小鼠大脑海马区神经元胞浆变性坏死、深染、核固缩、核空泡等，Nissl小体及正常神经元数量明显减少（P<0.05）。结论产黑普雷沃氏菌（Pn）加重了牙周炎的牙槽骨破坏，加速了小鼠海马区神经元细胞坏死及形态异常，并降低了小鼠记忆、学习等认知功能。

关键词: 产黑普雷沃氏菌, 口腔细菌, 牙周炎, 认知障碍, 行为学

Abstract:

Objective To investigate the effects of periodontitis induced by Prevotella nigrescens (Pn) combined with ligation on cognitive functions in mice. Methods Twenty-four C57BL/6J mice were randomly divided into control group, ligation group, and ligation + Pn treatment (P+Pn) group. Experimental periodontitis was induced by silk ligation of the first molars followed by topical application of Pn for 6 weeks. After modeling, alveolar bone resorption was assessed using micro-CT and histological analysis. Learning and memory abilities of the mice were evaluated using open field test (OFT), novel object recognition test (NORT), and Morris water maze test (MWM). Seven weeks after the start of modeling, the mice were sacrificed for examining histopathological changes in the hippocampus using HE and Nissl staining. Results After 6 weeks of molar ligation, micro-CT revealed horizontal alveolar bone resorption and furcation exposure in the mice, and histological analysis showed apical migration of the junctional epithelium, epithelial ridge hyperplasia, and lymphocyte infiltration, and these changes were obviously worsened in P+Pn group. Alveolar bone height decreased significantly in both ligation groups compared to the control group. Cognitive tests showed that the mice in both of the ligation groups traveled shorter distances in OFT, showed reduced novel object preference in NORT, and exhibited longer escape latencies in MWM, and the mice in P+Pn group had significantly poorer performances in the tests. Histologically, obvious neuronal cytoplasmic degeneration, necrosis, nuclear pyknosis, vacuolation, and reduced Nissl bodies and viable neurons were observed in the hippocampal regions of the mice in the two ligation groups. Conclusion Pn infection aggravates alveolar bone destruction, accelerates necrosis and causes morphological abnormalities of neuronal cells in the hippocampus to reduce cognitive functions of mice with periodontitis.

Key words: Prevotella nigrescens, oral bacteria, periodontitis, cognitive impairment, behavioral changes

陈琦, 夏天天, 周永强, 常铭洋, 胡楠, 杨燕美, 李仲, 高月, 顾斌. 产黑普雷沃氏菌加重小鼠的牙周炎症并损伤认知功能[J]. 南方医科大学学报, 2025, 45(3): 453-460.

Qi CHEN, Tiantian XIA, Yongqiang ZHOU, Mingyang CHANG, Nan HU, Yanmei YANG, Zhong LI, Yue GAO, Bin GU. Prevotella nigrescens exacerbates periodontal inflammation and impairs cognitive function in mice[J]. Journal of Southern Medical University, 2025, 45(3): 453-460.

图/表 6

图1 实验流程图

Fig.1 Flowchart of the experiment.

图2 建模6周小鼠牙周组织学切片HE染色

Fig.2 HE staining of periodontal histological sections of the mice at 6 weeks after modeling (scale bar=200 μm for upper and lower panels; P: Periodontitis).

图3 小鼠牙周炎模型建立及Micro-CT检测3D扫描分析

Fig.3 Establishment of a mouse periodontitis model and Micro-CT scan and 3D reconstruction analysis. A: Periodontitis in a mouse model. B: Micro-CT scan and 3D reconstruction for measurement of positional markings. C: Analysis of periodontal bone defects by Micro-CT and the distance from the enamel bone boundary to the top of the alveolar ridge (CAJ-ABC). **P<0.01 vs control group. P: Periodontitis.

图4 牙周炎模型小鼠观察期体质量变化

Fig.4 Body weight changes during the observation period in periodontitis mice. **P<0.01, ***P<0.001 vs control group; #P<0.05 vs P group.

图5 两种方法建立的牙周炎模型对小鼠学习记忆功能的影响

Fig.5 Learning and memory functions of mice with periodontitis modeled using the two methods. A: Open field test. B: representative tracking images of movement over 5 min, total distance covered, percent distance covered on the central grid, percent time spent on the central grid. C: New object recognition test (NORT). D: Percentage of distance covered around a new object, discrimination index (DI), percentage of times a new object was explored. E: Morris water maze test (MWM). F: Platform finding latency 5 days prior to training. G: Percentage of platform finding latency, number of platform traversals, time spent in target quadrant activity, and percentage of distance traveled in target quadrant activity. (*P<0.05,**P<0.01, ***P<0.001, ****P<0.0001 vs control group; #P<0.05, ##P<0.01, ####P<0.0001 vs P group. P: Periodontitis group; P+Pn: Periodontitis + Pn bacteria group).

图6 组织学观察牙周炎模型小鼠大脑海马神经元形态及功能的影响

Fig.6 Histological observations for assessing morphological and functional changes of hippocampal neurons in the brain of the mouse models. A: HE staining of the hippocampal region of the mice in each group (scale bar=200 μm) and the normal neuron numbers in the CA3 region. B: Nissl staining of the hippocampal region of the mice in each group (scale bar=200 μm) and Nissl-positive neuron numbers in the CA3 region. *P<0.05 vs control group; #P<0.05 vs P group.

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产黑普雷沃氏菌加重小鼠的牙周炎症并损伤认知功能

Prevotella nigrescens exacerbates periodontal inflammation and impairs cognitive function in mice

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