P. Gingivalis LPS通过TLR-4抑制Sema3A/Nrp1促进破骨细胞生成并损害成骨细胞分化

IF 3.7 3区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

International dental journal Pub Date : 2025-10-21 DOI:10.1016/j.identj.2025.103941

Jingyi Feng , Dongjie Fu , Sandeth Phan , Xiaoli Hu , Quan Xing

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引用次数: 0

摘要

目的：本研究表明，Sema3A/Nrp1通路对牙龈卟啉单胞菌LPS （P-LPS）诱导的骨溶解具有保护作用，并确定了其中的关键分子机制。方法：采用Real-time-PCR、western blot和ELISA检测P-LPS对破骨前细胞系（RAW267.4）和原代小鼠骨髓基质细胞（BMSCs）中Sema3A/Nrp1表达的影响。CCK8法检测重组Sema3A对P-LPS处理下RAW267.4细胞及BMSC增殖的影响。采用Real-time-PCR分析破骨细胞和成骨细胞标记基因。测定骨髓间充质干细胞成骨分化的ALP活性和矿化程度。测定破骨细胞分化的TRAP染色和活性。应用显微ct对p - lps诱导的颅骨溶骨模型进行分析。结果：P-LPS诱导破骨前细胞系（RAW267.4）和原代小鼠骨髓基质细胞（BMSCs）中Sema3A/Nrp1的表达均显著下调。此外，P-LPS暴露可提高破骨细胞基因表达，增加TRAP活性，促进rankl诱导的破骨细胞分化。然而，这些促进作用在给药Sema3A后减弱。此外，P-LPS还会损害骨髓间充质干细胞的成骨细胞分化，表现为成骨标志物的抑制、β-catenin转录活性的减弱和矿化程度的降低，这些都可以通过补充Sema3A来恢复。有趣的是，Sema3A减轻了p - lps诱导的骨髓间充质干细胞增殖抑制，但不影响RAW267.4细胞的增殖。此外，P-LPS通过toll样受体4 （TLR-4）下调Sema3A/Nrp1的表达。此外，一项体内研究显示，Sema3A可显著改善p - lps介导的炎症性骨溶解。结论：我们的数据表明Sema3A-Nrp1轴是p - lps敏感的骨稳态调节剂，通过双重调节破骨细胞和成骨细胞的活性，具有逆转感染诱导的骨溶解的治疗潜力。临床意义：我们的工作确定了上述双重靶向机制，为治疗感染相关口腔溶骨症的新疗法奠定了基础。

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P. Gingivalis LPS Drives Osteoclastogenesis and Impairs Osteoblast Differentiation by Suppressing Sema3A/Nrp1 Via TLR-4

Objectives

This study demonstrates that the Sema3A/Nrp1 pathway confers protection against Porphyromonas gingivalis LPS (P-LPS)-induced osteolysis and identifies key molecular mechanisms involved.

Methods

The effect of P-LPS on Sema3A/Nrp1 expression assessed in the preosteoclast cell line (RAW267.4) and primary mouse bone marrow stromal cells (BMSCs) using Real-time-PCR, western blot and ELISA. The effect of recombinant Sema3A on RAW267.4 cell and BMSC proliferation under P-LPS treatment was determined by CCK8 assay. Osteoclastic and osteoblastic markers gene was analysed by Real-time-PCR. ALP activity and mineralization were tested in BMSCs for osteoblast differentiation. TRAP staining and activity were measured for osteoclast differentiation. Micro-CT was applied to analysed P-LPS-induced calvarial osteolytic model.

Results

P-LPS induced significantly downregulated Sema3A/Nrp1 expression in both the preosteoclast cell line (RAW267.4) and primary mouse bone marrow stromal cells (BMSCs). Moreover, P-LPS exposure elevated osteoclastic gene expression, increased TRAP activity and promoted RANKL-induced osteoclast differentiation. However, these effects of promotion were attenuated by Sema3A administration. In addition, P-LPS impaired osteoblast differentiation in BMSCs, as evidenced by depressed osteogenic markers, attenuated β-catenin transcriptional activity, and diminished mineralization, all of which were rescued by Sema3A supplementation. Intriguingly, Sema3A alleviated P-LPS-induced repression of proliferation in BMSCs but did not affect the proliferation of RAW267.4 cells. Furthermore, P-LPS downregulated the expression of Sema3A/Nrp1 via Toll-like receptor 4 (TLR-4). Additionally, an in vivo study revealed that Sema3A administration markedly ameliorated P-LPS-mediated inflammatory osteolysis.

Conclusion

Our data identify the Sema3A-Nrp1 axis as a P-LPS–sensitive regulator of bone homeostasis, offering therapeutic potential to reverse infection-induced osteolysis through dual modulation of osteoclast and osteoblast activities. Sema3A attenuated PLPS- driven bone loss via concurrent osteoclast inhibition and osteoblast stimulation

Clinical Relevance

Our work identifies the above dual-targeting mechanism as a foundation for novel therapies against infection-related oral osteolysis.

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来源期刊

International dental journal 医学-牙科与口腔外科

CiteScore

4.80

自引率

6.10%

发文量

159

审稿时长

63 days

期刊介绍： The International Dental Journal features peer-reviewed, scientific articles relevant to international oral health issues, as well as practical, informative articles aimed at clinicians.