牙龈卟啉单胞菌脂多糖活化的巨噬细胞外泌体通过p38 MAPK/MMP-9信号通路促进破骨细胞分化

doi:10.13701/j.cnki.kqyxyj.2026.07.005

摘要/Abstract

摘要： 目的: 探究牙龈卟啉单胞菌脂多糖(Porphyromonas gingivalis-lipopolysaccharide, Pg-LPS)活化的巨噬细胞外泌体通过调控p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)/基质金属蛋白酶9(matrix metalloprotein 9,MMP-9)轴对破骨细胞分化的影响。方法: 丝线结扎法构建小鼠牙周炎模型,显微计算机断层扫描技术(micro-computed tomography,micro-CT)评估牙槽骨流失,实时荧光定量聚合酶链反应(quantitative reverse transcription polymerase chain reaction,qRT-PCR)和Western blot检测小鼠牙龈组织中CD11b、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、MMP-9、p38 MAPK的表达水平。Pg-LPS活化巨噬细胞,收集细胞培养上清(conditioned medium, CM),并提取外泌体(exosomes, exo),取CM/exo与破骨细胞前体细胞共培养。使用qRT-PCR和Western blot检测MMP-9、p38 MAPK表达水平,抗酒石酸酸性磷酸酶(Tartrate-resistant acid phosphatase,TRAP)染色评估破骨细胞分化情况。为进一步检测p38 MAPK通路在巨噬细胞外泌体促进破骨细胞分化中的作用,小鼠牙周采用丝线结扎联合局部注射外泌体,同时给予p38 MAPK通路抑制剂SB203580,检测骨丧失、MMP-9、p38 MAPK的表达水平;与CM/exo共培养的破骨细胞前体细胞中添加SB203580,检测MMP-9、p38 MAPK表达水平及破骨细胞分化情况。结果: 与对照组相比,丝线结扎小鼠牙槽骨丧失增加,牙龈组织中CD11b、IL-1β、IL-6、TNF-α、MMP-9、p38 MAPK表达水平升高;局部注射LPS-exo可促进牙周炎小鼠的骨流失,应用SB203580可缓解损伤。体外实验中,与对照组相比,LPS-CM/exo组MMP-9和p38 MAPK通路表达增加,TRAP染色见大量分化破骨细胞,SB203580可抑制LPS-CM/exo引起的破骨细胞分化作用(P<0.05)。结论: Pg-LPS活化的巨噬细胞外泌体通过激活p38 MAPK/MMP-9轴,促进破骨细胞分化,从而引起牙周炎骨丧失。

关键词: 牙龈卟啉单胞菌脂多糖, 巨噬细胞, 破骨细胞, 外泌体, p38丝裂原活化蛋白激酶

Abstract: Objective: To investigate whether exosomes derived from porphyromonas gingivalis-lipopolysaccharide (Pg-LPS)-activated macrophages promote osteoclasts differentiation via the p38 mitogen-activated protein kinase (p38 MAPK)/matrix metalloprotein 9 (MMP-9) signaling pathway. Methods: A mouse periodontitis model was established by ligature, and alveolar bone loss was assessed using micro-computed tomography (Micro-CT), quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were used to detect CD11b, interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), MMP-9, and p38 MAPK expression. Macrophages were stimulated with Pg-LPS, conditioned medium (CM) and exosomes (exo) were collected and co-cultured with osteoclast precursor cells. MMP-9 and p38 MAPK expression and osteoclast differentiation tartrate-resistant acid phosphatase (TRAP staining) were evaluated. The role of p38 MAPK was further tested by injecting exosomes locally with/without the p38 inhibitor SB203580, and by adding SB203580 to co-cultured precursors. Results: Ligature-induced periodontitis increased bone loss and elevated inflammatory and targeted molecule expression. In vitro, LPS-CM/exo enhanced MMP-9 and p38 MAPK expression and osteoclastogenesis, however, the effects were inhibited by SB203580 (P<0.05). Conclusion: Exosomes derived from Pg-LPS-activated macrophages promote osteoclast differentiation via the p38 MAPK/MMP-9 signaling pathway, which drove periodontal bone loss.

Key words: Porphyromonas gingivalis-lipopolysaccharide, macrophage, osteoclast, exosome, p38 mitogen-activated protein kinase

程立婷, 许若楠, 戚齐, 邢田. 牙龈卟啉单胞菌脂多糖活化的巨噬细胞外泌体通过p38 MAPK/MMP-9信号通路促进破骨细胞分化[J]. 口腔医学研究, 2026, 42(7): 575-583.

CHENG Liting, XU Ruonan, QI Qi, XING Tian. Exosomes Derived from Pg-LPS-activated Macrophages Promote Osteoclasts Differentiation via p38 MAPK/MMP-9 Signaling Pathway[J]. Journal of Oral Science Research, 2026, 42(7): 575-583.

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