The First Dongguan Affiliated Hospital, Guangdong Medical University, Dongguan, China.
Department of Oral and Maxillofacial Surgery & Special Dental Care University Medical Center Utrecht, Utrecht, The Netherlands.
J Periodontol. 2024 Jan;95(1):50-63. doi: 10.1002/JPER.23-0241. Epub 2023 Aug 18.
Protein lysine lactylation (Kla) has been proved to be closely related to inflammatory diseases, but its role in periodontitis (PD) is unclear. Therefore, this study aimed to establish the global profiling of Kla in PD models in rats.
Clinical periodontal samples were collected, the inflammatory state of tissues was verified by H&E staining, and lactate content was detected by a lactic acid kit. Kla levels were detected by immunohistochemistry (IHC) and Western blot. Subsequently, the rat model of PD was developed and its reliability verified by micro-CT and H&E staining. Mass spectrometry analysis was conducted to explore the expression profile of proteins and Kla in periodontal tissues. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed, and a protein-protein interaction (PPI) network was constructed. The lactylation in RAW264.7 cells was confirmed by IHC, immunofluorescence and Western blot. The relative expression levels of inflammatory factors IL-1β, IL-6, TNF-α, macrophage polarization-related factors CD86, iNOS, Arg1, and CD206 in RAW264.7 cells were detected by real time-quantitative polymerase chain reaction (RT-qPCR).
We observed substantial inflammatory cell infiltration in the PD tissues, and the lactate content and lactylation levels were significantly increased. The expression profiles of protein and Kla were obtained by mass spectrometry based on the established rat model of PD. Kla was confirmed in vitro and in vivo. After inhibiting the "writer" of lactylation P300 in RAW264.7 cells, the lactylation levels decreased, and the expression of inflammatory factors IL-1β, IL-6, and TNF-α increased. Meanwhile, the levels of CD86 and iNOS increased, and Arg1 and CD206 decreased.
Kla may play an important role in PD, regulating the release of inflammatory factors and polarization of macrophages.
蛋白赖氨酸乳酰化（Kla）已被证明与炎症性疾病密切相关，但它在牙周炎（PD）中的作用尚不清楚。因此，本研究旨在建立 PD 大鼠模型中 Kla 的全局分析。
收集临床牙周组织样本，通过 H&E 染色验证组织的炎症状态，通过乳酸试剂盒检测乳酸含量。通过免疫组织化学（IHC）和 Western blot 检测 Kla 水平。随后，建立 PD 大鼠模型，并通过 micro-CT 和 H&E 染色验证其可靠性。通过质谱分析探索牙周组织中蛋白质和 Kla 的表达谱。进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析，并构建蛋白质-蛋白质相互作用（PPI）网络。通过 IHC、免疫荧光和 Western blot 验证 RAW264.7 细胞中的乳酰化。通过实时定量聚合酶链反应（RT-qPCR）检测 RAW264.7 细胞中炎症因子 IL-1β、IL-6、TNF-α、巨噬细胞极化相关因子 CD86、iNOS、Arg1 和 CD206 的相对表达水平。
我们观察到 PD 组织中大量炎症细胞浸润，乳酸含量和乳酰化水平显著增加。基于建立的 PD 大鼠模型，通过质谱获得了蛋白质和 Kla 的表达谱。在体外和体内验证了 Kla。抑制 RAW264.7 细胞中乳酰化“写作者”P300 后，乳酰化水平降低，炎症因子 IL-1β、IL-6 和 TNF-α 的表达增加。同时，CD86 和 iNOS 水平升高，Arg1 和 CD206 水平降低。
Kla 可能在 PD 中发挥重要作用，调节炎症因子的释放和巨噬细胞的极化。
