白细胞介素-37通过抑制NLRP3炎性小体激活和调节M1/M2巨噬细胞极化来改善牙周炎的发展。
Interleukin-37 ameliorates periodontitis development by inhibiting NLRP3 inflammasome activation and modulating M1/M2 macrophage polarization.
作者信息
Yang Liyan, Tao Wei, Xie Chen, Chen Qiuye, Zhao Yunshan, Zhang Li, Xiao Xu, Wang Shilu, Zheng Xu
机构信息
Department of Stomatology, The First Affiliated Hospital of Hainan Medical University, Haikou, China.
School of Stomatology, Hainan Medical University, Haikou, China.
出版信息
J Periodontal Res. 2024 Feb;59(1):128-139. doi: 10.1111/jre.13196. Epub 2023 Nov 10.
OBJECTIVE
Our study was designed to explore the role of IL-37 in M1/M2 macrophage polarization imbalance in the pathogenesis of periodontitis.
BACKGROUND
Periodontitis is a chronic progressive inflammatory disease featured by gingival inflammation and alveolar bone resorption. Recent research has revealed that regulating macrophage polarization is a viable method to ameliorate periodontal inflammation. IL-37 is an anti-inflammatory cytokine, which has been reported to inhibit innate and adaptive immunity.
METHODS
For in vitro experiment, mouse macrophage RAW264.7 cells were pretreated with 0.1 ng/mL recombinant human IL-37. M1 and M2 polarizations of RAW264.7 cells were induced by 100 ng/mL LPS and 20 ng/mL IL-4, respectively. The expression of M1 (iNOS, TNF-α, and IL-6) and M2 (CD206, Arg1, and IL-10) phenotype markers in RAW264.7 cells was detected by RT-qPCR, western blotting, and immunofluorescence staining. For in vivo experiment, experimental periodontitis mouse models were established by sterile silk ligation (5-0) around the bilateral maxillary second molar of mice for 1 week. H&E staining of the maxillary alveolar bone was used to show the resorption of root cementum and dentin. Alveolar bone loss in mouse models was evaluated through micro-CT analysis. The expression of iNOS and CD206 in gingival tissues was assessed by immunohistochemistry staining. NLRP3 inflammasome activation was confirmed by western blotting.
RESULTS
IL-37 pretreatment reduced iNOS, TNF-α, and IL-6 expression in LPS-treated RAW264.7 cells but increased CD206, Arg1, and IL-10 in IL-4-treated RAW264.7 cells. LPS-induced upregulation in NLRP3, GSDMD, cleaved-IL-1β, and cleaved-caspase-1 expression was antagonized by IL-37 treatment. In addition, IL-37 administration ameliorated the resorption of root cementum and dentin in periodontitis mouse models. IL-37 prominently decreased iNOS+ cell population but increased CD206+ cell population in gingival tissues of periodontitis mice. The enhancement in NLRP3, GSDMD, cleaved-IL-1β, and cleaved-caspase-1 expression in the gingival tissues of periodontitis mice was offset by IL-37 administration.
CONCLUSION
IL-37 prevents the progression of periodontitis by suppressing NLRP3 inflammasome activation and mediating M1/M2 macrophage polarization.
目的
本研究旨在探讨白细胞介素-37（IL-37）在牙周炎发病机制中M1/M2巨噬细胞极化失衡中的作用。
背景
牙周炎是一种以牙龈炎症和牙槽骨吸收为特征的慢性进行性炎症性疾病。最近的研究表明，调节巨噬细胞极化是改善牙周炎症的一种可行方法。IL-37是一种抗炎细胞因子，据报道可抑制先天性和适应性免疫。
方法
体外实验中，小鼠巨噬细胞RAW264.7细胞用0.1 ng/mL重组人IL-37预处理。RAW264.7细胞的M1和M2极化分别由100 ng/mL脂多糖（LPS）和20 ng/mL白细胞介素-4（IL-4）诱导。通过逆转录定量聚合酶链反应（RT-qPCR）、蛋白质印迹法和免疫荧光染色检测RAW264.7细胞中M1（诱导型一氧化氮合酶（iNOS）、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6））和M2（甘露糖受体C型1（CD206）、精氨酸酶1（Arg1）和白细胞介素-10（IL-10））表型标志物的表达。体内实验中，通过在小鼠双侧上颌第二磨牙周围用无菌丝线（5-0）结扎1周建立实验性牙周炎小鼠模型。上颌牙槽骨的苏木精-伊红（H&E）染色用于显示牙骨质和牙本质的吸收。通过显微计算机断层扫描（micro-CT）分析评估小鼠模型中的牙槽骨丢失。通过免疫组织化学染色评估牙龈组织中iNOS和CD206的表达。通过蛋白质印迹法确认NLRP3炎性小体的激活。
结果
IL-37预处理降低了LPS处理的RAW264.7细胞中iNOS、TNF-α和IL-6的表达，但增加了IL-4处理的RAW264.7细胞中CD206、Arg1和IL-10的表达。IL-37处理拮抗了LPS诱导的NLRP3、Gasdermin D（GSDMD）、裂解的白细胞介素-1β（cleaved-IL-1β）和裂解的半胱天冬酶-1（cleaved-caspase-1）表达上调。此外，给予IL-37改善了牙周炎小鼠模型中牙骨质和牙本质的吸收。IL-37显著减少了牙周炎小鼠牙龈组织中iNOS+细胞群，但增加了CD206+细胞群。给予IL-37抵消了牙周炎小鼠牙龈组织中NLRP3、GSDMD、cleaved-IL-1β和cleaved-caspase-1表达的增强。
结论
IL-37通过抑制NLRP3炎性小体激活和介导M1/M2巨噬细胞极化来阻止牙周炎的进展。
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