Jiang Yiming, Wang Zihan, Zhang Kaige, Hu Yue, Shang Dehao, Jiang Lulu, Huang Minghao, Wang Biyao, He Xiaomin, Wu Zhou, Yan Xu, Zhang Xinwen
The VIP Department, School and Hospital of Stomatology, China Medical University, Liaoning Provincial Key Laboratory of Oral Diseases, Shenyang, China.
Department of Oral Implantology, School and Hospital of Stomatology, China Medical University, Liaoning Provincial Key Laboratory of Oral Diseases, Shenyang, China.
J Clin Periodontol. 2025 Apr;52(4):622-636. doi: 10.1111/jcpe.14111. Epub 2025 Jan 6.
To explore the potential roles of mitochondrial dysfunction in the initiation of inflammation in periodontal macrophages and to determine the mechanism underlying the involvement of dynamin-related protein 1 (Drp1) in macrophage inflammatory responses through its interaction with hexokinase 1 (HK1).
Gingival tissues were collected from patients diagnosed with periodontitis or from healthy volunteers. Drp1 tetramer formation and phosphorylation were analysed using western blot. THP-1 macrophages and RAW264.7 cells were stimulated with Porphyromonas gingivalis (Pg) or Pg lipopolysaccharide (Pg LPS), respectively. Alterations in proteins associated with mitochondrial dynamics were scrutinized via western blot. Immunofluorescence was used to evaluate mitochondrial damage and mitochondrial permeability transition pore (mPTP) opening. Western blot was used to examine the inflammatory markers NLRP3, caspase-1, IL-1β and GSDMD. Protein interactions involving Drp1 were verified through immunoprecipitation.
In periodontitis patient samples, Pg LPS-treated RAW264.7 cells, and Pg-stimulated THP-1 macrophages, over-activated Drp1 was able to drive NLRP3 inflammasome activation and the subsequent release of inflammatory factors. A direct interaction between Drp1 and HK1 was observed, facilitating excessive mPTP opening and subsequent mitochondrial dysfunction.
In the inflammatory milieu of periodontal tissues, Drp1 hyperactivation in the macrophages is implicated in inflammation induction. Modulation of the inflammatory response in periodontal macrophages by Drp1 appears to facilitate mPTP opening.
探讨线粒体功能障碍在牙周巨噬细胞炎症起始中的潜在作用，并确定动力相关蛋白1（Drp1）通过与己糖激酶1（HK1）相互作用参与巨噬细胞炎症反应的潜在机制。
从诊断为牙周炎的患者或健康志愿者中采集牙龈组织。采用蛋白质免疫印迹法分析Drp1四聚体的形成和磷酸化情况。分别用牙龈卟啉单胞菌（Pg）或Pg脂多糖（Pg LPS）刺激THP-1巨噬细胞和RAW264.7细胞。通过蛋白质免疫印迹法仔细观察与线粒体动力学相关蛋白质的变化。采用免疫荧光法评估线粒体损伤和线粒体通透性转换孔（mPTP）开放情况。用蛋白质免疫印迹法检测炎症标志物NLRP3、半胱天冬酶-1、白细胞介素-1β和Gasdermin D（GSDMD）。通过免疫沉淀验证涉及Drp1的蛋白质相互作用。
在牙周炎患者样本、经Pg LPS处理的RAW264.7细胞和经Pg刺激的THP-1巨噬细胞中，过度激活的Drp1能够驱动NLRP3炎性小体激活及随后炎症因子的释放。观察到Drp1与HK1之间存在直接相互作用，促进了mPTP过度开放及随后的线粒体功能障碍。
在牙周组织的炎症环境中，巨噬细胞中Drp1的过度激活与炎症诱导有关。Drp1对牙周巨噬细胞炎症反应的调节似乎促进了mPTP开放。
