State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, China.
Applied Oral Sciences and Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong, China.
Int Immunopharmacol. 2024 Mar 10;129:111658. doi: 10.1016/j.intimp.2024.111658. Epub 2024 Feb 15.
Chronic periodontitis triggers an increase in osteoclastogenesis, with glycolysis playing a crucial role in this process. Pyruvate kinase M2 (PKM2) is a critical enzyme involved in glycolysis and pyruvate metabolism. Yet, the precise function of PKM2 in osteoclasts and their formation remains unclear and requires further investigation.
Bioinformatics was used to investigate critical biological processes in osteoclastogenesis. In vitro, osteoclastogenesis was analyzed using tartrate-resistant acid phosphatase (TRAP) staining, phalloidin staining, quantitative real‑time PCR (RT-qPCR), and Western blotting. Small interfering RNA (siRNA) of PKM2 and Shikonin, a specific inhibitor of PKM2, were used to verify the role of PKM2 in osteoclastogenesis. The mouse model of periodontitis was used to assess the effect of shikonin on bone loss. Analyses included micro computed tomography, immunohistochemistry, flow cytometry, TRAP staining and HE staining.
Bioinformatic analysis revealed a significant impact of glycolysis and pyruvate metabolism on osteoclastogenesis. Inhibition of PKM2 leads to a significant reduction in osteoclastogenesis. In vitro, co-culture of the heat-killed Porphyromonas gingivalis significantly promoted osteoclastogenesis, concomitant with an increased PKM2 expression in osteoclasts. Shikonin weakened the promoting effect of porphyromonas gingivalis on osteoclastogenesis. In vivo experiments demonstrated that inhibition of PKM2 by shikonin alleviated bone loss induced by periodontitis, suppressed excessive osteoclastogenesis in alveolar bone, and reduced tissue inflammation to some extent.
PKM2 inhibition by shikonin, a specific inhibitor of this enzyme, attenuated osteoclastogenesis and bone resorption in periodontitis. Shikonin appears to be a promising therapeutic agent for treating periodontitis.
慢性牙周炎会引发破骨细胞生成增加，糖酵解在此过程中起着关键作用。丙酮酸激酶 M2（PKM2）是糖酵解和丙酮酸代谢中的关键酶。然而，PKM2 在破骨细胞及其形成中的确切功能尚不清楚，需要进一步研究。
使用生物信息学方法研究破骨细胞生成中的关键生物学过程。在体外，通过抗酒石酸酸性磷酸酶（TRAP）染色、鬼笔环肽染色、实时定量 PCR（RT-qPCR）和 Western blot 分析破骨细胞生成。使用 PKM2 的小干扰 RNA（siRNA）和 PKM2 的特异性抑制剂紫草素验证 PKM2 在破骨细胞生成中的作用。使用牙周炎小鼠模型评估紫草素对骨丢失的影响。分析包括微计算机断层扫描、免疫组织化学、流式细胞术、TRAP 染色和 HE 染色。
生物信息学分析显示糖酵解和丙酮酸代谢对破骨细胞生成有显著影响。抑制 PKM2 会导致破骨细胞生成显著减少。在体外，热灭活牙龈卟啉单胞菌共培养显著促进破骨细胞生成，同时破骨细胞中的 PKM2 表达增加。紫草素减弱了牙龈卟啉单胞菌对破骨细胞生成的促进作用。体内实验表明，紫草素抑制 PKM2 减轻了牙周炎引起的骨丢失，抑制了牙槽骨中破骨细胞的过度生成，并在一定程度上减轻了组织炎症。
牙龈卟啉单胞菌的特异性抑制剂紫草素抑制 PKM2 可减轻牙周炎中的破骨细胞生成和骨吸收。紫草素似乎是治疗牙周炎的一种有前途的治疗剂。
