The State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
The First Affiliated Hospital of Chengdu Medical College, Chengdu, China.
Oral Dis. 2024 May;30(4):2485-2496. doi: 10.1111/odi.14693. Epub 2023 Jul 30.
To investigate the involvement and role of receptor-interacting protein 3 (RIP3)-mediated necroptosis in periodontitis.
A periodontitis murine model was established by oral infection with Porphyromonas gingivalis, and activation of necroptosis pathway was identified by immunohistochemistry. Adeno-associated virus was used to knock down Rip3 and the effect of Rip3 knockdown on periodontal inflammation was examined by Micro-CT, qRT-PCR and histological staining. In vitro, P. gingivalis-LPS was used to infect fibroblast cell line L929 and siRNA was used to knock down Rip3. Necroptosis pathway signalling and inflammation in cells were detected by cell viability and death assay, Western Blot, qRT-PCR and immunofluorescence analysis.
Phosphorylation of RIP3 and mixed lineage kinase domain-like protein (MLKL) was increased in the periodontal ligament of mice infected with P. gingivalis. RIP3 knockdown reduced osteoclastogenesis and inflammatory cytokines in the periodontal area, and alleviated alveolar bone loss in vivo. In vitro, P. gingivalis-LPS-induced RIP3-mediated necroptosis in L929 cells, and knockdown of RIP3 by siRNA decreased the expression of inflammatory cytokines.
RIP3-mediated necroptosis is activated in periodontitis and blocking necroptosis alleviates disease progression, indicating that RIP3 may be a potential target for periodontitis treatment.
研究受体相互作用蛋白 3(RIP3)介导的坏死性凋亡在牙周炎中的作用。
通过口腔感染牙龈卟啉单胞菌建立牙周炎小鼠模型,通过免疫组织化学鉴定坏死性凋亡途径的激活。使用腺相关病毒敲低 Rip3,并通过 Micro-CT、qRT-PCR 和组织学染色检测 Rip3 敲低对牙周炎炎症的影响。体外使用牙龈卟啉单胞菌脂多糖(P. gingivalis-LPS)感染成纤维细胞系 L929,并使用 siRNA 敲低 Rip3。通过细胞活力和死亡检测、Western Blot、qRT-PCR 和免疫荧光分析检测细胞中坏死性凋亡途径信号和炎症。
牙龈卟啉单胞菌感染小鼠牙周韧带中 RIP3 和混合谱系激酶结构域样蛋白(MLKL)的磷酸化增加。RIP3 敲低减少了牙周区域的破骨细胞生成和炎症细胞因子,并减轻了体内牙槽骨丢失。体外,P. gingivalis-LPS 诱导 L929 细胞中 RIP3 介导的坏死性凋亡,siRNA 敲低 RIP3 降低了炎症细胞因子的表达。
RIP3 介导的坏死性凋亡在牙周炎中被激活,阻断坏死性凋亡可减轻疾病进展,表明 RIP3 可能是牙周炎治疗的潜在靶点。