华西口腔医学杂志

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6~8岁儿童龋病相关唾液蛋白组的电喷雾离子肼—串联质谱分析

Yan Guowei1,2, Huang Wenming1,2, Xue Honglei1,2, Jia Yuhuan1,2, Yang Deqin1,2   

  1. 1. Dept. of Endodontics, The Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing 400017, China; 2. Chongqing Key Laboratory of Oral Diseases and Biomedical Science, Chongqing 400017, China
  • 出版日期:2014-06-01 发布日期:2014-06-01
  • 通讯作者: 杨德琴,教授,博士,E-mail:yangdeqin@gmail.com
  • 作者简介:鄢国伟,硕士,E-mail:yanw1109@126.com
  • 基金资助:

    国家自然科学基金资助项目(81170950);重庆市卫生局医学科研计划重点基金资助项目(2011-1-062);重庆市医学重点学科《牙体牙髓病学》建设基金资助项目(2011);重庆市渝北区科技基金资助项目[(2011)33号]

Relationship between dental caries and salivary proteome by electrospray ionization ion-trap tandem mass spectro-metry in children aged 6 to 8 years

1.重庆医科大学附属口腔医院牙体牙髓科;2.口腔疾病与生物医学重庆市重点实验室,重庆 400017   

  1. 1. Dept. of Endodontics, The Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing 400017, China; 2. Chongqing Key Laboratory of Oral Diseases and Biomedical Science, Chongqing 400017, China
  • Online:2014-06-01 Published:2014-06-01

摘要:

目的  采用电喷雾离子肼—串联质谱分析法(ESI-MS/MS)比较高龋组和无龋组儿童唾液蛋白组,初步探索唾液蛋白组与龋病的关系,并探索龋敏感性相关的生物标志物。方法  以高龋组、无龋组儿童各10名为研究对象,收集唾液样本,进行聚丙烯酰胺凝胶电泳(SDS-PAGE),经过滤-辅助的蛋白酶解(FASP)及液相柱色谱分析后,进行ESI-MS/MS鉴定,并对高龋唾液组和无龋唾液组组间的蛋白差异进行分析。结果  高龋组蛋白量显著高于无龋组,高龋组、无龋组鉴定的多肽数分别为602、481个,分别属于286、227个蛋白。两组之间差异表达多肽数为361个,差异表达蛋白数为118个,包含基质金属蛋白酶9、黏蛋白7、乳铁蛋白、碳酸酐酶6、天青杀素、冷凝集素等。结论  高龋组儿童唾液蛋白量高于无龋组,基质金属蛋白酶9、黏蛋白7、乳铁蛋白、碳酸酐酶6、天青杀素、冷凝集素等高龋组与无龋组之间差异表达蛋白的检测,为进一步探索龋敏感性相关生物蛋白标志物奠定了基础。

关键词: 龋病, 唾液蛋白组, 电喷雾离子肼—串联质谱, 生物标志物

Abstract:

Objective  To explore the relationship between salivary proteome and dental caries and to promote the bio-marker studies of dental caries susceptibility by comparing the salivary proteome of caries-active children and caries-free children with electrospray ionization ion-trap tandem mass spectrometry (ESI-MS/MS). Methods  Ten caries-active children and ten caries-free children were sampled. The salivary proteome of the two groups was studied, and the differential protein between the two groups was analyzed by ESI-MS/MS after sodium dodecyl sulfate polyacrylamide gel electrophoresis, filter-aided sample preparation, and liquid chromatography. Results  The concentration of salivary protein was higher in the caries-active group than in the caries-free group. The polypeptide counts of thecaries-active and caries-free groups were 602 and 481, which belonged to 286 and 227 proteins, respectively. The differential polypeptide count of the two groups was 361, and the differential protein count was 118. The detected proteins included matrix metalloproteinase-9 (MMP9), mucin-7 (MUC7), lactotransferrin (LTF), carbonic anhydrase 6 (CA6), azurocidin (AZU), and cold agglutinin. Conclusion  The total salivary protein was higher in the caries-active group than in the caries-free group. The preliminary detection of differential proteins (MMP9, MUC7, LTF, CA6, AZU, and cold agglutinin) may lay some foundation for biomarker research of dental caries susceptibility.

Key words: dental caries, salivary proteome, elec-trospray ionization ion-trap tandem mass spectrometry, biomarkers