基于同位素标记相对和绝对定量技术的良性气道狭窄肉芽组织成纤维细胞蛋白质组学分析  

Isobaric tags for relative and absolute quantitation based proteomics profiling of fibroblasts in granulation tissue of benign airway stenosis

作　　者：罗钰龙 王颖治[1] 陈小波[1] 吴宏楷[1] 朱易平 钟长镐[1] 陈愉[1] 李时悦[1] Luo Yulong;Wang Yingzhi;Chen Xiaobo;Wu Hongkai;Zhu Yiping;Zhong Changhao;Chen Yu;Li Shiyue(Fifth Affiliated Hospital of Guangzhou Medical University,Innovation Centre for Advanced Interdisciplinary Medicine,Key Laboratory of Biological Targeting Diagnosis,Therapy and Rehabilitation of Guangdong Higher Education Institutes,Guangzhou 510700,China;First Affiliated Hospital of Guangzhou Medical University,National Center for Respiratory Disease,Guangzhou Institute of Respiratory Health,State Key Laboratory of Respiratory Diseases in Guangzhou,National Clinical Medical Center for Respiratory Diseases,Guangzhou 510120,China)

机构地区：[1]广州医科大学附属第五医院、前沿医学交叉研究中心、广东高校生物靶向诊治与康复重点实验室,广州10700,广州医科大学附属第一医院、国家呼吸医学中心、广州呼吸健康研究院、呼吸疾病国家重点实验室、国家呼吸系统疾病临床医学中心,广州510120

出　　处：《中华生物医学工程杂志》2022年第4期361-367,共7页Chinese Journal of Biomedical Engineering

基　　金：广东省高校特色创新团队(2021KCXTD028);广州市基础研究计划(202102020916)。

摘　　要：目的对良性气道狭窄(BAS)患者肉芽组织成纤维细胞进行蛋白质组学分析, 以进一步揭示BAS的发病机制。方法取5例BAS患者的肉芽组织(BAS组)和3例肺癌患者的正常支气管组织(对照组)进行成纤维细胞原代培养, 用同位素标记相对和绝对定量(iTRAQ)技术分析成纤维细胞的蛋白质组表达情况, 筛选差异表达蛋白, 并使用基因本体(GO)数据库、京都基因和基因组(KEGG)数据库和重复出现邻近基因检索工具(STRING)数据库对差异表达蛋白的功能、代谢通路和蛋白相互关系进行富集分析。结果共筛选出93个差异表达蛋白。其中有36个差异表达蛋白在BAS肉芽组织成纤维细胞中显著下调, 57个差异表达蛋白显著上调。差异表达蛋白主要定位于细胞膜和细胞外区域, 参与离子结合功能、代谢活性功能, 主要参与的通路有糖酵解-糖异生代谢、细胞外基质-受体结合、磷脂酰肌醇3-激酶(PI3K)-AKT信号传导、复杂环境中的微生物代谢和次级代谢物合成。结论 BAS狭窄部位肉芽组织成纤维细胞的细胞外基质-受体结合异常、PI3K-AKT信号通路异常和代谢异常可能与BAS的发生发展有关。Objective To perform the proteomics profiles of fibroblasts in granulation tissue from patients with benign airway stenosis(BAS)and thereby investigate the pathogenesis of BAS.Methods Granulation tissue from 5 patients with BAS(BAS group)and normal bronchial tissue from 3 patients with lung cancer(control group)were harvested for primary culture of fibroblasts.The profile of proteome expression was examined with isobaric tags for relative and absolute quantitation(iTRAQ)to screen for differentially expressed proteins(DEPs).The Gene Ontology(GO)database,Kyoto Encyclopedia of Genes and Genomes(KEGG)database and Search Tool for Recurring Instances of Neighboring Genes(STRING)database were used for enrichment analysis of the functions,metabolic pathways and interactions of the differentially expressed proteins.Results A total of 93 DEPs were screened,among them,36 were significantly down-regulated,and 57 were significantly up-regulated,in fibroblasts of granulation tissue of BAS.The DEPs were mainly localized on cell membrane and in the extracellular domain,and were engaged in ion binding and metabolic activity.The major pathways involved were glycolysis-gluconeogenesis metabolism,extracellular matrix-receptor binding,phosphatidylinositol 3-kinase(PI3K)-AKT signaling,microbial metabolism and secondary metabolite synthesis in complex environments.Conclusion Abnormalities in extracellular matrix-receptor binding,PI3K-AKT signaling pathway and metabolism in fibroblasts of granulation tissue in lesion of BAS may be related to occurrence and development of BAS.

关 键 词：蛋白质组 细胞外基质 信号通路 代谢 良性气道狭窄 

分 类 号：R56[医药卫生—呼吸系统]