牙龈卟啉单胞菌脂多糖刺激人牙髓干细胞炎症相关蛋白质组学研究

doi:10.13701/j.cnki.kqyxyj.2022.08.008

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (8): 736-741.DOI: 10.13701/j.cnki.kqyxyj.2022.08.008

牙龈卟啉单胞菌脂多糖刺激人牙髓干细胞炎症相关蛋白质组学研究

谢元栋^1,2, 李泽华^1,2, 刘晓莉^1,2, 胡玲^1,2, 詹骆宁^1,2, 李毅^1*

1.吉林大学口腔医院儿童口腔科吉林长春 130021;
2.吉林大学牙发育与颌骨重塑吉林省重点实验室吉林长春 130021

收稿日期:2021-12-10 出版日期:2022-08-28 发布日期:2022-08-24
通讯作者: *李毅,E-mail:lyi99@jlu.edu.cn
作者简介:谢元栋(1995~ ),男,山东泰安人,硕士在读,研究方向:牙髓病学、牙源性干细胞的应用。
基金资助:
吉林省自然科学基金(编号:20200201409JC)吉林省财政厅科技项目(编号:JCSZ2020304-6)

Proteomic Analysis of Inflammation-related Differential Proteins in Human Dental Pulp Stem Cells Induced by Porphyromonas Gingivalis Lipopolysaccharide

XIE Yuandong^1,2, LI Zehua^1,2, LIU Xiaoli^1,2, HU Ling^1,2, ZHAN Luoning^1,2, LI Yi^1*

1. Department of Pediatric Dentistry, School of Stomatology, Jilin University, Changchun 130021, China;
2. Jilin Provincial Key Laboratory of Tooth Development and Bone Remodeling, Changchun 130021, China

Received:2021-12-10 Online:2022-08-28 Published:2022-08-24

摘要/Abstract

摘要： 目的: 采用蛋白质组学的方法探究牙龈卟啉单胞菌脂多糖(P.gingivalis LPS)诱导人牙髓干细胞(DPSCs)炎症相关差异蛋白的表达。方法: 体外分离培养并鉴定人DPSCs,利用10 μg/mL P.gingivalis LPS刺激DPSCs建立炎症模型,实时荧光定量聚合酶链反应(PCR)和酶联免疫吸附试验(ELISA)检测炎症因子白细胞介素(IL)-6和IL-8的表达量。利用蛋白质组学的方法筛选P.gingivalis LPS诱导DPSCs炎症相关的差异表达蛋白,通过KEGG、GO和PPI进行功能富集分析。结果: P.gingivalis LPS刺激DPSCs导致炎症因子IL-6和IL-8的表达显著升高。与对照组相比,DPSCs中有277种差异表达蛋白在炎症微环境下发生了显著变化,功能富集分析结果显示差异表达蛋白参与调控多条炎症或损伤修复相关的信号通路如中性粒细胞介导的免疫过程、肿瘤坏死因子介导信号通路、凋亡信号通路、Wnt通路、TGF-β信号通路、磷酸戊糖途径、HIF-1信号通路等生物过程。结论: P.gingivalis LPS可诱导DPSCs发生炎症反应,此过程涉及多蛋白、多信号通路的相互联系,为探索牙髓炎的发病机制及临床诊断标记物的研究奠定了基础。

关键词: 牙龈卟啉单胞菌, 脂多糖, 牙髓干细胞, 蛋白质组学, 生物信息学

Abstract: Objective: To investigate the expression of inflammation-related differential proteins in human dental pulp stem cells (DPSCs) induced by Porphyromonas gingivalis lipopolysaccharide (P.gingivalis LPS) by proteomics. Methods: Human DPSCs were isolated, cultured, and identified in vitro. Ten μg/mL P.gingivalis LPS was used to stimulate DPSCs to establish an inflammatory model. The expressions of inflammatory factors interleukin-6 (IL-6) and IL-8 were detected by real-time fluorescence quantitative polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). The differentially expressed proteins related to inflammation induced by P.gingivalis LPS were screened by proteomics, and the functional enrichment was analyzed by KEGG, GO, and PPI. Results: P.gingivalis LPS stimulated the expression of inflammatory factors IL-6 and IL-8 in DPSCs. Compared with the control group, 277 differentially expressed proteins (DEPs) in DPSCs were changed significantly in the inflammatory microenvironment. Functional enrichment analysis showed that DEPs were involved in regulating multiple inflammatory or damage repair related signal pathways, such as neutrophil mediated immune process, tumor necrosis factor-mediated signal pathway, apoptosis signal pathway, Wnt pathway, TGF-β biological processes such as signal pathway, pentose phosphate pathway, and HIF-1 signal pathway. Conclusion: P.gingivalis LPS can induce an inflammatory response in DPSCs. This process involves the relationship between multiple proteins and multiple signal pathways, which lays a foundation for exploring the pathogenesis of pulpitis and the study of clinical diagnostic markers.

Key words: porphyromonas gingivalis, lipopolysaccharide, dental pulp stem cells, proteomics, bioinformatics

谢元栋, 李泽华, 刘晓莉, 胡玲, 詹骆宁, 李毅. 牙龈卟啉单胞菌脂多糖刺激人牙髓干细胞炎症相关蛋白质组学研究[J]. 口腔医学研究, 2022, 38(8): 736-741.

XIE Yuandong, LI Zehua, LIU Xiaoli, HU Ling, ZHAN Luoning, LI Yi. Proteomic Analysis of Inflammation-related Differential Proteins in Human Dental Pulp Stem Cells Induced by Porphyromonas Gingivalis Lipopolysaccharide[J]. Journal of Oral Science Research, 2022, 38(8): 736-741.

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牙龈卟啉单胞菌脂多糖刺激人牙髓干细胞炎症相关蛋白质组学研究

Proteomic Analysis of Inflammation-related Differential Proteins in Human Dental Pulp Stem Cells Induced by Porphyromonas Gingivalis Lipopolysaccharide

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