基于NANO-LC-MS/MS和生物信息学的抗特发性肺纤维化靶点的筛选与验证  

Screening and Validation of Anti-idiopathic Pulmonary Fibrosis Targets Based on NANO-LC-MS/MS and Bioinformatics

作　　者：宋蔚 吴柳莹[1,2] 张文婷 胡晓艺 杜晓月 王梦雅 郑雪瑶 左佩文 彭志红 SONG Wei;WU Liu-ying;ZHANG Wen-ting;HU Xiao-yi;DU Xiao-yue;WANG Meng-ya;ZHENG Xue-yao;ZUO Pei-wen;PENG Zhi-hong(School of Life Sciences,Hubei University,Wuhan 430062,China;National and Local Joint Engineering Research Center of High-throughput Drug Screening Technology,Hubei University,Wuhan 430062,China)

机构地区：[1]湖北大学生命科学学院,武汉430062 [2]湖北大学药物高通量筛选技术国家地方联合工程研究中心,武汉430062

出　　处：《中国药学杂志》2022年第20期1733-1741,共9页Chinese Pharmaceutical Journal

基　　金：药物高通量筛选技术国家地方联合工程研究中心开放基金项目资助(M20202006;K20202001)。

摘　　要：目的利用生物信息技术检索特发性肺纤维化(idiopathic pulmonary fibrosis,IPF)基因芯片数据,并结合体外实验,筛选与验证IPF相关的蛋白,为该疾病的药物开发提供潜在的作用靶点。方法采用基因表据库(Gene Expression Omnibus,GEO)检索“IPF”词条,下载GSE150910芯片数据,利用Bioconductor软件包DESeq2分析正常组和IPF组差异表达基因,对所获得差异基因进行基因本体论(gene ontology,GO)功能分析、日本京都基因与基因组百科全书(kyoto gene and genome encyclopedia,KEGG)通路分析、蛋白互作网络分析并作可视化处理。通过液质联用对差异基因对应的蛋白质进行定量;体外研究关键蛋白对α平滑肌肌动蛋白(α-SMA)分泌或形成的影响。结果共筛选到69个与细胞外基质形成相关的差异表达基因,包括53个上调基因和16个下调基因;功能分析显示,差异基因主要涉及到细胞外基质的降解(degradation of the extracellular matrix)、胶原蛋白降解(collagen degradation)和胶原链三聚化(collagen chain trimerization)通路;蛋白互作网络分析和蛋白定量研究显示,与IPF相关的基因主要为金属蛋白酶(MMP-3,9,ADAMTS14)和P4HA3等;金属蛋白酶抑制剂和P4HA3抑制剂能抑制肺纤维α-SMA分泌。结论通过筛选差异表达基因,明确相关基因和蛋白在IPF发生或发展过程中作用,为IPF新药研发提供新思路和方案。OBJECTIVE To retrieve idiopathic pulmonary fibrosis(IPF)gene chip data using bioinformatics technology and provide potential target proteins for development of drugs for IPF combined with in vitro assay.METHODS The Gene Expression Omnibus(GEO)was searched by the“idiopathic pulmonary fibrosis”entry.The GSE150910 microarray data was downloaded and the difference in the expressed genes between the normal group and the IPF group were analyzed using the Bioconductor package DESeq2.Differential genes were obtained for analysis of GO function,KEGG pathway,and protein interaction network and then underwent visualization treatment.The proteins corresponding to the differential genes were quantified by liquid chromatography-mass spectrometry.The effects of key proteins onα-SMA secretion or formation were studied in vitro.RESULTS A total of 69 differentially expressed genes were screened,including 53 up-regulated genes and 16 down-regulated genes;functional analysis showed that the main differential genes were related with extracellular matrix degradation,collagen degradation,and collagen chain trimerization pathways;protein interaction network analysis and protein quantification studies showed that the genes associated with IPF were mainly metalloproteinases(MMP-3,9,ADAMTS14)and P4HA3,etc.;matrix metalloproteinase inhibitors and P4HA3 inhibitors could inhibit the secretion ofα-SMA.CONCLUSION By screening differentially expressed genes.The roles of related genes and proteins can be clarified in the occurrence or development of IPF and provide new ideas and solutions for the development of new drugs for IPF.

关 键 词：特发性肺纤维化 生物信息技术 细胞外基质 纳升级液质联用 

分 类 号：R966[医药卫生—药理学]