Inhibition of receptor‐interacting protein kinase‐3 in the necroptosis pathway attenuates inflammatory bone loss in experimental apical periodontitis in Balb/c mice

作者：Jie Liu, Jiajia Wang, Jie Ren, Qin Yang, Weicheng Zhan, Min Wang, Liang Hao, Yuan Yue · 发表于：International Endodontic Journal · 年份：2021 · DOI：10.1111/iej.13534 · 被引用次数：19 · 研究领域：Bone Metabolism and Diseases、Cell death mechanisms and regulation、Oral microbiology and periodontitis research

AIM: To explore the role of necroptosis in apical periodontitis (AP), this study investigated necroptosis in a Fusobacterium nucleatum (Fn)-induced AP model of Balb/c mice and explored related intracellular signalling pathways in L929 cells affected by Fn. METHODOLOGY: For the in vivo experiments, expression of receptor-interacting protein kinase-3 (RIP3) was inhibited using an adeno-associated virus and then the Balb/c mice model of AP was established by injecting Fn into the root canal of the first mandibular molars. Bone loss and number of osteoclasts were measured via micro-computed tomography and tartrate-resistant acid phosphatase staining, respectively; expression of RIP3 and phosphorylated mixed lineage kinase domain-like protein (pMLKL) was detected by immunohistochemistry and western blotting; expression of mRNA of inflammatory cytokines was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). For the in vitro experiments, L929 cells transfected with RIP3-Mus-siRNA or negative control siRNA were co-cultured with Fn; thereafter, western blotting, detection of cell death and viability and qRT-PCR analyses were performed to assess the activation of necroptosis pathway and expression of mRNA of inflammatory cytokines. Data were analysed with unpaired t-test and one-way analysis of variance with significance set at p < .05. RESULTS: The Fn-induced apical lesions were associated with apical bone loss, an increased number of osteoclasts, enhanced exp...