基于外周血单个核细胞转录组测序结果的硅沉着病发病机制研究  

Study on silicosis mechanism by transcriptome sequencing of peripheral blood mononuclear cells

作　　者：李海斌 张林 关毅 赵英政 李宁 刘楠 王永恒 周强 常美玉 吴卫东 姚三巧 LI Haibin;ZHANG Lin;GUAN Yi;ZHAO Yingzheng;LI Ning;LIU Nan;WANG Yongheng;ZHOU Qiang;CHANG Meiyu;WU Weidong;YAO Sanqiao(Department of Labor Health and Environmental Hygiene,School of Public Health,North China University of Science and Technology,Tangshan,Hebei 063200;Department of Labor Health and Environmental Hygiene,School of Public Health,Xinxiang Medical University,Xinxiang,Henan 453003;Shandong Provincial Maternal and Child Health Care Hospital,Jinan 250001)

机构地区：[1]华北理工大学公共卫生学院劳动卫生与环境卫生学教研室,河北唐山063200 [2]新乡医学院公共卫生学院劳动卫生与环境卫生学教研室,河南新乡453003 [3]山东省妇幼保健院,济南250001

出　　处：《郑州大学学报（医学版）》2024年第6期782-787,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(U21A20334);河南省高等教育教学改革研究与实践项目(研究生教育类)(2023SJGLX061Y)。

摘　　要：目的:筛选硅沉着病患者外周血单个核细胞中的差异表达基因,并分析其相关调控通路,探索硅沉着病的发病机制。方法:采集3例硅沉着病患者和3名健康对照空腹静脉血,分离外周血单个核细胞,采用高通量测序技术和生物信息学分析技术分析差异表达基因和可能信号通路,采用蛋白质-蛋白质互作网络筛选中枢基因。选取常规体检的117名工人作为验证人群,分为健康对照组、接尘对照组和硅沉着病组,采用RT-qPCR技术检测3组研究对象外周血单个核细胞Ⅰ型胶原、Ⅲ型胶原、磷酸肌醇3-激酶(PI3K)、整合素β1(ITGβ1)和整合素连接激酶(ILK)mRNA表达水平。结果:转录组数据分析检测到80813个mRNA;limma测试共获得1915个差异表达基因,其中987个表达上调,928个表达下调。蛋白质-蛋白质互作网络分析提示差异表达基因聚焦到3个中枢基因:PI3K、ITGβ1和ILK。基因集富集分析证实,PI3K、ITGβ1和ILK可能通过Janus激酶/信号转导子和转录激活子(JAK-STAT)信号通路、Wnt信号通路和细胞黏附分子信号通路参与硅沉着病发生发展。与健康对照组比较,接尘对照组和硅沉着病组外周血单个核细胞中Ⅰ型胶原、Ⅲ型胶原、PI3K、ITGβ1和ILK mRNA表达水平升高,且硅沉着病组Ⅰ型胶原、PI3K、ITGβ1 mRNA表达水平更高(P<0.05)。结论:PI3K、ITGβ1和ILK基因可能通过JAK-STAT、Wnt和细胞黏附分子信号通路参与硅沉着病发生发展过程。Aim:To screen differentially expressed genes in peripheral blood mononuclear cells of patients with silicosis,and analyze their related regulatory pathways,so as to provide new ideas and methods for the study of the pathogenesis of silicosis.Methods:Fasting venous blood was collected from 3 silicosis patients and 3 healthy controls.Peripheral blood mononuclear cells were isolated,and differential expressed genes and possible signaling pathways were analyzed using high-throughput sequencing technology and bioinformatics analysis technology.PPI analysis was used to screen central genes.A total of 117 workers who underwent routine physical examinations were selected as the validation population.They were allocated into healthy control group,dust exposed control group,and silicosis group.RT-qPCR technology was used to detect the expression levels of typeⅠcollagen,typeⅢcollagen,phosphoinositide 3-kinase(PI3K),integrinβ1(ITGβ1),and integrin-linked protein kinase(ILK)genes in peripheral blood mononuclear cells of the study subjects in the 3 groups.Results:Totally 80813 mRNAs were detected;limma test results indicated that 1915 differentially expressed genes were obtained,out of which,987 were upregulated and 928 were downregulated.PPI analysis suggested that these differentially expressed genes focused on 3 central genes,PI3K,ITGβ1,and ILK.Gene set enrichment analysis confirmed that PI3K,ITGβ1,and ILK might be closely related to the possible mechanism of silicosis through the JAK-STAT,Wnt,and cell adhesion molecule signaling pathways.Compared with the healthy control group,the mRNA expression levels of typeⅠcollagen,typeⅢcollagen,PI3K,ITGβ1,and ILK genes in peripheral blood mononuclear cells of the dust exposed control group and silicosis group were all increased,and those of typeⅠcollagen,PI3K,and ITGβ1 were higher in the silicosis group(P<0.05).Conclusion:PI3K,ITGβ1,and ILK genes may be involved in the possible mechanism of silicosis through the JAK-STAT,Wnt,and cell adhesion molecule signaling pathwa

关 键 词：硅沉着病 外周血单个核细胞 磷酸肌醇3-激酶 整合素Β1 整合素连接激酶 

分 类 号：R135.2[医药卫生—劳动卫生]