【摘要】 目的研究正畸牙移动中前扣带回皮质层(ACC)的蛋白质组学表达,筛选牙移动疼痛相关的目标蛋白,探讨ACC在疼痛传导和形成中的作用。方法 30只雄性SD大鼠建立实验性牙移动模型后,根据行为学观测结果分组。实验组进行加力,对照组仅进行乙醚麻醉,24 h后进行行为学观测并取ACC组织,根据行为学观测结果选取4标样本进行蛋白质组学研究。ACC组织在提取蛋白并胰酶消化后,经串联质谱标记(TMT)试剂标记,进行液相色谱-串联质谱(LC-MS/MS)检测,并对差异表达蛋白进行生物信息学分析,进行基因本体(GO)注释,GO功能聚类分析,基因和基因组学京都百科全书(KEGG)代谢通路注释及功能聚类分析。结果本实验共鉴定并定量3374种蛋白,其中表达上调蛋白12种、表达下调蛋白109种。GO注释和功能聚类分析中从生物过程、细胞组成、分子功能三个方面阐述了大鼠在实验性牙移动应力下发生的一系列应激反应及代谢调节。KEGG注释及功能聚类分析表明与炎症相关通路花生四烯酸通路及JAK-STAT信号通路表达下调,且差异具有统计学意义。花生四烯酸通路的前列腺素E合成酶(PGES)、谷胱甘肽过氧化物酶1(GPX1)、环氧化物水解酶2(EPHX2),JAK-STAT信号通路的信号转导和转录因子1(STAT1)、STAT2显著下调,可能与炎症性疼痛的发生、发展密切相关。结论 TMT技术是组织蛋白质组学研究的有效方法。ACC在牙移动疼痛形成和调节中发挥重要作用,其中花生四烯酸通路、JAK-STAT信号通路可能是牙移动疼痛的关键信号通路。本实验为进一步探讨牙移动疼痛关键蛋白提供了重要的实验依据。更多还原

【Abstract】 Objective To illucidate the proteomic expression in the anterior cingulate cortex(ACC)after experimental tooth movement and screening the significant proteins related with orthodontic pain,further investigate the role ACC plays in orthodontic pain. Methods In this research,30 male Sprague-Dawley rats(250 ~ 300 g)were applied with orthodontic appliance. They were divided into two groups and selected for further experiment according to behavior assessment. After tandem mass tag(TMT)labeling,the protein extracted from ACC was detected with liquid chromatography-tandem mass spectrometry(LC-MS/MS). Then bioinformatic analysis of the differentially expressed proteins was carried out,including Gene Ontology(GO)annotation,GO functional enrichment clustering,and Kyoto Encyclopedia of Genes and Genomes(KEGG)annotation and functional enrichment clustering. Results In total,3374 proteins were identified and quantified,among which 109 proteins were significantly down-regulated,while 12 proteins were up-regulated. GO annotation and functional enrichment clustering showed the posttraumatic stress response and biochemical metabolism regulation happened in ACC following orth-odontic tooth movement stress from three categories:biological process,cellular component and molecular function. KEGG annotation and functional enrichment clustering results showed that Arachidonic acid metabolism and JAK-STAT signaling pathway which were relevant to inflammatory pain were significantly down-regulated. 5 proteins were found closely related with the inflammary orthodontic pain,including prostaglandin E synthase(PGES),glutathione peroxidase 1(GPX1),bifunctional epoxide hydrolase 2(EPHX2)in the Arachidonic acid metabolism,as well as signal transducer and activator of transcription 1(STAT1),signal transducer and activator of transcription 2(STAT2)in the JAK-STAT signaling pathway.Conclusions TMT technology was proved an effective method evaluating proteomic expression. ACC plays a significant role in the orthodontic pain. Arachidonic acid metabolism and JAK-STAT signaling pathway are possible critical signaling pathways involved in tooth movement.更多还原