通过 necrostatin-1 调节 RIP1 介导的坏死性凋亡在牙周炎中的作用。

Regulation of RIP1-Mediated necroptosis via necrostatin-1 in periodontitis.

机构信息

Department of Prosthodontics, The State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.

Department of Prosthodontics, Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction, Tianjin Stomatological Hospital, School of Medicine, Nankai University, Tianjin, China.

出版信息

J Periodontal Res. 2023 Oct;58(5):919-931. doi: 10.1111/jre.13150. Epub 2023 Jun 19.

DOI:10.1111/jre.13150

PMID:37334934

Abstract

OBJECTIVE

To explore the mechanism of receptor-interacting protein 1 (RIP1)-mediated necroptosis during periodontitis progression.

BACKGROUND

RIP3 and mixed lineage kinase domain-like protein (MLKL) have been detected to be upregulated in periodontitis models. Because RIP1 is involved in necroptosis, it might also play a role in the progression of periodontitis.

METHODS

An experimental periodontitis model in BALB/c mice was established by inducing oral bacterial infection. Western blotting and immunofluorescence analyses were used to detect RIP1 expression in the periodontal ligament. Porphyromonas gingivalis was used to stimulate L929 and MC3T3-E1. RIP1 was inhibited using small-interfering RNA. Western blotting, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA) analyses were used to detect the effect of necroptosis inhibition on the expression of damage-associated molecular patterns and inflammatory cytokines. Necrostatin-1 (Nec-1) was intraperitoneally injected to inhibit RIP1 expression in mice. Necroptosis activation and inflammatory cytokine expression in periodontal tissue were verified. Tartrate-resistant acid phosphatase staining was applied to observe osteoclasts in the bone tissues of different groups.

RESULTS

RIP1-mediated necroptosis was activated in mice with periodontitis. P. gingivalis induced RIP1-mediated necroptosis in L929 and MC3T3-E1 cells. After RIP1 inhibition, the expression levels of high mobility group protein B1 (HMGB1) and inflammatory cytokines were downregulated. After inhibiting RIP1 with Nec-1 in vivo, necroptosis was also inhibited, the expression levels of HMGB1 and inflammatory cytokines were downregulated, and osteoclast counts in the periodontal tissue decreased.

CONCLUSION

RIP1-mediated necroptosis plays a role in the pathological process of periodontitis in mice. Nec-1 inhibited necroptosis, alleviated inflammation in periodontal tissue, and reduced bone resorption in periodontitis.

摘要

目的

探索受体相互作用蛋白 1（RIP1）在牙周炎进展过程中介导坏死性凋亡的机制。

背景

在牙周炎模型中已检测到 RIP3 和混合谱系激酶结构域样蛋白（MLKL）上调。由于 RIP1 参与坏死性凋亡，因此它可能在牙周炎的进展中也发挥作用。

方法

通过诱导口腔细菌感染，建立 BALB/c 小鼠实验性牙周炎模型。使用 Western blot 和免疫荧光分析检测牙周韧带中 RIP1 的表达。使用牙龈卟啉单胞菌刺激 L929 和 MC3T3-E1。使用小干扰 RNA 抑制 RIP1。使用 Western blot、逆转录定量聚合酶链反应（RT-qPCR）和酶联免疫吸附测定（ELISA）分析检测坏死性凋亡抑制对损伤相关分子模式和炎症细胞因子表达的影响。腹腔内注射 Necrostatin-1（Nec-1）抑制小鼠 RIP1 的表达。验证牙周组织中坏死性凋亡的激活和炎症细胞因子的表达。应用抗酒石酸酸性磷酸酶染色观察不同组骨组织中的破骨细胞。

结果

在牙周炎小鼠中，RIP1 介导的坏死性凋亡被激活。牙龈卟啉单胞菌诱导 L929 和 MC3T3-E1 细胞中 RIP1 介导的坏死性凋亡。抑制 RIP1 后，高迁移率族蛋白 B1（HMGB1）和炎症细胞因子的表达水平下调。体内用 Nec-1 抑制 RIP1 后，坏死性凋亡也被抑制，HMGB1 和炎症细胞因子的表达水平下调，牙周组织中的破骨细胞计数减少。