RAC2-MAPK14-HSPA8途径参与肺结核性肉芽肿形成的机制  

Mechanism of RAC2-MAPK14-HSPA8 pathway involved in the formation of pulmonary granuloma

作　　者：李奇凤 徐振华 刘志刚[2] 赵奕威 Li Qifeng;Xu Zhenhua;Liu Zhigang;Zhao Yiwei(Xinjiang Institute of Pediatrics,Children′s Hospital of Xinjiang Uygur Autonomous Region,Xinjiang Hospital of Beijing Children′s Hospital,Urumqi 830054,China;Department of Thoracic Surgery,the Eighth Affiliated Hospital of Xinjiang Medical University,Urumqi 830049,China)

机构地区：[1]北京儿童医院新疆医院,新疆维吾尔自治区儿童医院,新疆儿科研究所,乌鲁木齐830054 [2]新疆医科大学第八附属医院胸外科,乌鲁木齐830049

出　　处：《国际呼吸杂志》2023年第12期1436-1442,共7页International Journal of Respiration

基　　金：国家自然科学基金(81960387);新疆维吾尔自治区自然科学基金(2021D01E33)。

摘　　要：目的借助蛋白质组学对肺结核患者的病灶区(肉芽肿区)和非病灶区组织进行差异分析,挑选出关键基因,阐明肺结核性肉芽肿形成的作用机制。方法选取2020年10月至2021年2月在新疆医科大学第八附属医院行手术切除病灶的肺结核患者15例,均取肉芽肿病灶区(病例组)和非病灶区(对照组)的组织标本,其中3例进行蛋白提取检测、蛋白酶解除盐、同位素标记、质谱检测、数据搜索等最终形成蛋白质组学库。对筛选出来的差异蛋白进行基因本体注释(GO)、京都基因和基因组百科全书(KEGG)和互作网络分析等,蛋白印迹法对筛选出核心蛋白进行验证。结果差异蛋白火山图筛选出上调表达蛋白有65个,下调表达蛋白有151个;蛋白互作网络筛选出排名前10种蛋白,显著上调的为CTSD、HSPA8、ARHG、RAC2蛋白,显著下调的为MYH14、TJP1、CTNNB1、PXN、HSD17B6、RHOF蛋白。蛋白印迹法结果显示,HSPA8、MAPK14、RAC2在肺组织病灶区均明显增高,差异均有统计学意义(t值分别为3.20、3.23、6.30,均P<0.05),并参与抗原加工和呈递。结论本研究筛选出与肺结核性肉芽肿关系密切的排名前10种蛋白CTSD、HSPA8、ARHG、RAC2、MYH14、TJP1、CTNNB1、PXN、HSD17B6、RHOF,其中RAC2-MAPK14-HSPA8途径参与抗原加工和呈递,这在肉芽肿形成过程中扮演重要角色,也为研发抗结核药物提供了潜在治疗靶点。Objective To analyze the differences between lesion area(granuloma area)and non-lesion area tissues of pulmonary tuberculosis patients with the help of proteomics,to select the key genes,and to elucidate the mechanism of pulmonary tuberculous granuloma formation.Methods Fifteen patients with pulmonary tuberculosis undergoing surgery in the Eighth Affiliated Hospital of Xinjiang Medical University from October 2020 to February 2021 were resected with focal area(case group)and non-focal area(control group)of granuloma.Three of them were detected by protein extraction,protease desalting,isotope labeling,mass spectrometry detection,data search,etc.,finally forming a proteomics library.The screened differential proteins were analyzed by Gene Ontology Annotation(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG),and interaction network analysis,and the core proteins were verified by western blotting.Results The differential protein volcano map screened 65 up-regulated proteins and 151 down-regulated proteins,and the top 10 proteins were screened out by the protein interaction network,in which CTSD,HSPA8,ARHG,and RAC2 proteins were significantly upregulated and MYH14,TJP1,CTNNB1,PXN,HSD17B6,and RHOF proteins were significantly down-regulated.Western blotting showed that HSPA8,MAPK14,and RAC2 were significantly increased in the lesion area of lung tissue,with significant differences(t values were 3.20,3.23,and 6.30,respectively,all P<0.05),and they were involved in antigen processing and presentation.Conclusions The top 10 proteins CTSD,HSPA8,ARHG,RAC2,MYH14,TJP1,CTNNB1,PXN,HSD17B6,and RHOF that are closely related to pulmonary tuberculous granulomas were screened,among which RAC2-MAPK14-HSPA8 pathway was involved in antigen processing and presentation,which plays an important role in the formation of granulomas and provides potential therapeutic targets for the development of anti-tuberculosis drugs.

关 键 词：结核 蛋白质组学 肺组织 肉芽肿病灶区 非病灶区 

分 类 号：R521[医药卫生—内科学]