[1]张小军　尹　林　刘思远　张丽军.iTRAQ技术检测烟雾病病人硬脑膜特异性蛋白表达[J].中国临床神经外科杂志,2021,26(04):261-265.[doi:10.13798/j.issn.1009-153X.2021.04.012]
　Zhang Xiao-jun,YIN Lin,LIU Si-yuan,et al.Detection of specific protein expression of dural tissues in patients with moyamoya disease using iTRAQ technique[J].,2021,26(04):261-265.[doi:10.13798/j.issn.1009-153X.2021.04.012]
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iTRAQ技术检测烟雾病病人硬脑膜特异性蛋白表达()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
26
期数:
2021年04期
页码:
261-265
栏目:
实验研究
出版日期:
2021-04-25

文章信息/Info

Title:
Detection of specific protein expression of dural tissues in patients with moyamoya disease using iTRAQ technique
文章编号:
1009-153X（2021）04-0261-05
作者:
张小军　尹　林　刘思远　张丽军
200331 上海，上海天佑医院神经外科（张小军）；201508 上海，上海市公共卫生临床中心新药临床研究中心（尹　林、张丽军）；200052 上海，中国人民解放
Author(s):
Zhang Xiao-jun1; 3;  YIN Lin2;  LIU Si-yuan4;  ZHANG Li-jun2.
1. Department of Neurosurgery, Shanghai Tianyou Hospital, Shanghai 200331, China; 2. Department of Clinical Research Center, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China; 3. Department of Neurosurgery, The 905th Hospital of The Chinese People’s Liberation Army Navy, Shanghai 200052, China; 4. The College of Information, Mechanical and Electrical Engineering, Shanghai Normal University, Shanghai 201400, China
关键词:
烟雾病; 硬脑膜; 蛋白质组; 生物信息学; 组织芯片; 同位素相对标记与绝对定量技术（iTRAQ）
Keywords:
Moyamoya disease;  Dura mater;  Proteomics;  Bioinformatics;  Tissue microarray;  isobaric tag for relative and absolute
分类号:
R 743; Q 786
DOI:
10.13798/j.issn.1009-153X.2021.04.012
文献标志码:
A
摘要:
目的 优化硬脑膜蛋白质的提取方法，筛选烟雾病（MMD）病人硬脑膜特异性表达蛋白质。方法 收集96例MMD和13例非MMD病人的硬脑膜组织，用胶原酶Ⅳ孵育去除硬脑膜表面的血液，用机械破碎或液氮研磨方法提取蛋白质，一维凝胶电泳分离后进行质谱鉴定，用同位素相对标记与绝对定量（iTRAQ）技术进行标记、分离和鉴定差异蛋白质，应用R语言和生物信息学分析筛选MMD相关蛋白质，再应用组织芯片技术验证差异蛋白质表达。结果 胶原酶Ⅳ消化、液氮研磨和十二烷基硫酸钠法提取硬脑膜蛋白质效果较好；iTRAQ技术和生信分析发现23个差异蛋白质，组织芯片结果证实MMD病人硬脑膜组织SLC4A1蛋白表达明显下调。结论 iTRAQ技术可以很好地进行标记、分离和鉴定硬脑膜组织差异蛋白质，SLC4A1可能在MMD发病过程中具有一定的作用。
Abstract:
Objective To optimize the extraction method of dural proteins and screen the specific expression proteins of dural tissues in the patients with moyamoya disease (MMD) using isobaric tag for relative and absolute quantitation (iTRAQ) technique. Methods The dural tissues of 96 MMD and 13 non-MMD patients were collected, and incubated with collagenase Ⅳ to remove the blood on the surface of dura. The dural proteins were extracted by mechanical disruption and liquid nitrogen grinding, respectively; separated by one-dimensional gel electrophoresis, and identified by mass spectrometry. The differential expression proteins were labelled, separated and identified using iTRAQ technique, and then the R language and bioinformatics analysis were used to screen MMD-related proteins, and tissue microarray technique was used to verify the MMD-related proteins. Results The dural proteins could be effectively extracted using collagenase Ⅳ digestion, liquid nitrogen grinding and sodium lauryl sulfate extraction. Twenty-three differential expression proteins were screened and identified by the iTRAQ technique and biosynthesis analysis. Tissue microarray results verified that the protein expression of SLC4A1 was significantly down-regulated in the dural tissue of MMD patients. Conclusions iTRAQ technique can well label, separate and identify the differential expression proteins in the dural tissue. SLC4A1 may play certain role in the pathogenesis of MMD.

参考文献/References:

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备注/Memo

备注/Memo:
基金项目：十三五重大新药创制课题（2017ZX09304027）军海军第九〇五医院神经外科（张小军）；201400 上海，上海师范大学信息与机电工程学院（刘思远）
通讯作者：张丽军，E-mail：zhanglijun1221@163.com
更新日期/Last Update: 2021-04-25