香芹酚通过磷脂酰肌醇3-激酶途径诱导牙周炎牙槽骨修复的机制研究

doi:10.7518/hxkq.2025.2025050

摘要/Abstract

摘要：

目的本研究旨在探究香芹酚（CV）在调节牙周炎中牙槽骨修复作用的机制。方法通过结扎第一磨牙并注射脂多糖（LPS）建立大鼠牙周炎模型（P组），并设立对照组（C组）。治疗模型分别接受低剂量（L组）、中剂量（M组）和高剂量（H组）的CV水凝胶。采用苏木精-伊红染色观察牙周组织的病理变化。采用免疫组织化学染色分析Ⅰ型胶原（COL1）和runt相关转录因子2（Runx2）的表达。在体外实验中，将大鼠成骨细胞分为C、P、L、M、H、CV和CV+LY294002（CV+LY）组。通过Western blot分析检测牙龈组织及体外成骨细胞磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/糖原合成酶激酶3β（GSK-3β）通路相关蛋白和成骨蛋白。采用定量逆转录聚合酶链反应测定炎症因子和成骨蛋白基因的表达。使用碱性磷酸酶（ALP）显色试剂盒和茜素红S染色试剂盒评估成骨能力。通过免疫荧光（IF）检测成骨细胞中COL1的表达情况。透射电子显微镜检测细胞凋亡情况。结果 CV水凝胶可缓解牙周症状，上调PI3K/AKT/GSK-3β通路相关蛋白和成骨蛋白的表达，增加ALP表达和钙化结节数量。同时它降低了细胞凋亡和炎症因子的水平。LY294002抑制PI3K/AKT通路，并降低成骨蛋白表达、ALP染色和钙化结节数量。结论 CV水凝胶通过激活PI3K通路并抑制炎症诱导的骨吸收，促进牙槽骨成骨细胞的增殖和分化。本研究强调了CV在治疗牙周炎方面的潜力。

关键词: 香芹酚, 牙周炎, 成骨细胞, 磷脂酰肌醇3-激酶, 脂多糖

Abstract:

Objective This study aims to explore the mechanism of carvacrol (CV) in regulating alveolar bone repair in periodontal disease. Methods A rat model of periodontal disease (P group) was created by ligating the first molar and injecting lipopolysaccharide (LPS). A control model (C group) was also created. The treatment models received low (L group), medium (M group), and high (H group) doses of CV hydrogel. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in periodontal tissues. Immunohistochemical staining was employed to analyze the expression of collagen typeⅠ(COL1) and Runt-related transcription factor 2 (Runx2). In vitro, the rat osteoblast cells were divided into C, P, L, M, H, CV and CV+LY294002 (CV+LY) groups. Western blot analysis detected the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase 3 beta (GSK-3β) pathway-related and osteoblastic proteins. Quantitative reverse-transcription po-lymerase chain reaction was used to measure the expression of inflammatory factors and osteoblastic proteins. The alkaline phosphatase (ALP) colorimetric kit and alizarin red S staining kit were utilized to assess osteogenic ability. Immunofluorescence (IF) was used to detect COL1 expression in osteoblasts. Transmission electron microscopy was applied to detect cell apoptosis. Results CV hydrogel alleviated periodontal symptoms, upregulated PI3K/AKT/GSK-3β pathway-related and osteoblastic proteins, and increased the expression of ALP and the number of calcified nodules. However, it decreased cell apoptosis and inflammatory factors. LY294002 inhibited the PI3K/AKT pathway and decreased osteoblastic protein expression, ALP coloration, and calcified nodule quantity. Conclusion CV hydrogel promotes the proliferation and differentiation of alveolar bone osteoblasts by activating the PI3K pathway and inhibiting inflammation-induced bone resorption. This study emphasizes the potential of CV for the treatment of periodontal diseases.

Key words: carvacrol, periodontal disease, osteoblasts, phosphatidylinositol 3-kinase, lipopolysaccharide

中图分类号:

R781.4

滕念, 李兆勋, 高甜甜, 项艳蕊, 周露露, 高翔. 香芹酚通过磷脂酰肌醇3-激酶途径诱导牙周炎牙槽骨修复的机制研究[J]. 华西口腔医学杂志, 2026, 44(1): 94-105.

Teng Nian, Li Zhaoxun, Gao Tiantian, Xiang Yanrui, Zhou Lulu, Gao Xiang. Mechanism underlying carvacrol-induced alveolar bone repair via the phosphatidylinositol 3-kinase pathway in periodontal disease[J]. West China Journal of Stomatology, 2026, 44(1): 94-105.

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NCBI Reference Sequence	基因	引物（5’-3’）	长度/bp
XM_006253120.5	ALP	F：TGCACGACAATCGGGATGAA	20/20
		R：TAGTAACCGGAGCCCTCACA
NM_053304.1	COL1	F：GGTCTTGGTGGTTTTGTATTCG	22/21
		R：AACAGTCGCTTCACCTACAGC
XM_017596552.3	Runx2	F：TGGCCGGGAATGATGAGAAC	20/20
		R：GCTCCGGCCTACAAATCTCA
XM_063272870.1	OPN	F：CCAGCCAAGGACCAACTACA	20/20
		R：AGTGTTTGCTGTAATGCGCC
NM_031512.2	IL-1β	F：GCTTCCTTGTGCAAGTGTC	19/19
		R：TGGACAGCCCAAGTCAAGG
NM_012589.2	IL-6	F：GTTTCTCTCCGCAAGAGACTTC	22/20
		R：TGTGGGTGGTATCCTCTGTG
XM_063287055.1	COX-2	F：CGCAAATACTGCAGCCCTAC	20/20
		R：GGATGTCTTGTTCGTCTGCC
NM_012675.3	TNF-α	F：GGGTGATCGGTCCCAACAAG	20/19
		R：GCTTGGTGGTTTGCTACGA
NM_031144.3	β-actin	F：CTATCGGCAATGAGCGGTTCC	21/22
		R：GCACTGTGTTGGCATAGAGGTC

组别	SBI	PD/mm
C	0.60±0.52	0.21±0.05
P	2.80±0.42^A	0.90±0.17^A
L	2.22±0.44^a	0.76±0.13^A
M	1.91±0.54	0.69±0.15^a
H	1.20±0.63^b	0.41±0.15^B

组别	COL1	Runx2
C	95.30±4.45^b	96.35±2.35^b
P	11.81±4.48	17.98±2.03
L	34.14±7.61	73.61±8.21
M	63.76±2.64	89.32±7.70
H	72.17±5.27^b	96.34±3.67^b