Impact of chronic restraint stress-induced anxiety on periodontal tissue destruction in mice with experimental periodontitis
10.19405/j.cnki.issn1000–1492.2026.07.013
- VernacularTitle:慢性束缚应激诱导的焦虑对牙周炎小鼠牙周组织的影响
- Author:
Jian LIU
1
;
Junji XU
2
;
Yuanyin WANG
1
Author Information
1. College & Hospital of Stomatology， Anhui Medical University， Anhui Provincial Key Laboratory of Oral Diseases Research， Hefei 230032
2. Department of Periodontics， The Affiliated Stomatological Hospital of Chongqing Medical University， Chongqing 401147
- Publication Type:Journal Article
- Keywords:
chronic restraint stress;
anxiety;
periodontitis;
glucocorticoids;
neutrophils;
neutrophil extracellular traps
- From:
Acta Universitatis Medicinalis Anhui
2026;61(7):1253-1260
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the impact of chronic restraint stress-induced anxiety-like behavior on the pathological progression of experimental periodontitis in mice. MethodsA total of 40 C57BL/6J mice were randomly divided into two groups： normal control （Ctrl） and chronic restraint stress （CRS） （n=20）. Anxiety-like behavior was induced by 7 consecutive days of chronic restraint stress. Behavioral assessments were conducted using the open field test （OFT） and elevated plus maze （EPM） on days 8 and 9， respectively. Following behavioral evaluation， 10 mice from each group were randomly selected and subjected to experimental periodontitis by silk ligature placement at the cervical region of the right maxillary second molar. The final four experimental groups （n=10） were： normal control （Ctrl）， periodontitis alone （PD）， chronic restraint stress alone （CRS）， and chronic restraint stress with periodontitis （CP）. All mice were euthanized on day 7 post-periodontitis induction. Alveolar bone destruction was quantified by micro-computed tomography （micro-CT）. Histological analyses， including hematoxylin-eosin （HE）， Masson's trichrome， and tartrate-resistant acid phosphatase （TRAP） staining， were performed to evaluate alveolar bone resorption， periodontal fiber arrangement， and osteoclast numbers. Gingival neutrophil infiltration was analyzed by flow cytometry. Neutrophil extracellular traps （NETs） in gingival tissue and serum corticosterone （CORT） levels were measured by enzyme-linked immunosorbent assay （ELISA）. Additionally， the glucocorticoid receptor antagonist mifepristone （RU-486） was administered to investigate the role of glucocorticoid signaling in the pathogenesis of anxiety-aggravated periodontitis. ResultsChronic restraint stress successfully induced anxiety-like behavior in mice and exacerbated periodontitis progression. Micro-CT and histological analyses revealed that， compared with the PD group， the CP group exhibited aggravated alveolar bone resorption， more severe periodontal fiber destruction， and increased osteoclast numbers. Flow cytometry demonstrated significantly elevated gingival neutrophil proportions in the CP group relative to the PD group. ELISA analysis revealed significantly elevated NETs expression in gingival tissues and markedly increased serum CORT levels in the CP group compared to the PD group. RU-486 intervention effectively downregulated NETs expression in gingival tissues and attenuated the progression of anxiety-aggravated periodontitis. ConclusionAnxiety induced by chronic restraint stress activates the hypothalamic-pituitary-adrenal axis， promotes glucocorticoid release， subsequently upregulates neutrophil extracellular trap formation， and ultimately exacerbates alveolar bone destruction and tissue inflammation in experimental periodontitis.
