基于xCell算法研究痛风的免疫生物标志物及相关靶向中药的筛选  

Study on immune biomarker of gout and screening of related target Chinese medicine based on the xCell algorithm

作　　者：李玲琴[1] 周睿姣 张燕妮 袁心柱 LI Lingqin;ZHOU Ruijiao;ZHANG Yanni;YUAN Xinzhu(Department of Rheumatology and Immunology,The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Neurology,The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Nephrology,The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Nephrology,The Affiliated Nanchong Central Hospital of North Sichuan Medical College,Nanchong 637003,Sichuan Province,China)

机构地区：[1]川北医学院附属医院风湿免疫科,四川南充637000 [2]川北医学院附属医院神经内科,四川南充637000 [3]川北医学院附属医院肾内科,四川南充637000 [4]川北医学院附属南充市中心医院肾内科,四川南充637003

出　　处：《中国药师》2024年第6期1007-1018,共12页China Pharmacist

基　　金：四川省自然科学基金青年基金项目(2023NSFSC1577);川北医学院项目(CBY21-QA10);南充市科技局项目(19SXHZ0141)。

摘　　要：目的研究痛风致病的关键基因以及免疫生物标志物,挖掘潜在的靶向中药,为痛风的临床治疗提供新方向。方法从GEO数据库下载痛风芯片数据集,使用R软件进行差异基因的筛选,并对这些差异基因进行GO及基因通路KEGG富集分析。应用STRING数据库对差异基因进行蛋白质互作网络分析,利用Cytoscape筛选关键基因。使用ELISA方法对关键基因的蛋白表达水平进行实验验证。进一步使用xCell估计免疫细胞在痛风中的相对表达情况及相关性,最后通过Coremine Medical数据库对显著富集的免疫相关生物学过程及关键基因中药进行预测。结果筛选出痛风差异基因共计852个。在GO富集分析中,与免疫反应密切相关的主要生物学过程包括白细胞的趋化性、IL(白细胞介素)-2的产生以及血管生成的调控;KEGG通路主要富集于IL-17信号通路、趋化因子信号通路和肿瘤坏死因子(TNF)信号通路等。筛选出TNF、IL-6、IL-1β、趋化因子配体8(CXCL8)、FOS原癌基因(FOS)和血管内皮生长因子A(VEGFA)等10个关键蛋白,实验验证关键基因蛋白表达水平与芯片数据的表达趋势一致。免疫分析显示,单核细胞、记忆B细胞、初始CD8+T细胞和树突状细胞与痛风关系密切。中药预测发现,人参、三七、黄芩、三白草和丹参等为治疗痛风的潜在药物。结论本研究揭示了痛风致病的可能关键基因以及免疫机制,挖掘出潜在的靶向中药。这些关键基因和中药有望为痛风的治疗提供新的思路和方法。Objective To study the key genes and immune biomarkers of gout and to explore potential target Chinese medicine,which can provide new directions for the clinical treatment of gout.Methods The gout microarray dataset was downloaded from the GEO database,the differential genes were screened using R software,and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on these differential genes.The STRING database was applied to analyze the protein interaction network of the differential genes,and the Cytoscape was used to screen the key genes.The protein expression levels of key genes were validated using the ELISA method.Furthermore,the xCell was used to estimate the relative expression and correlation of immune cells in gout.Finally,the significantly enriched immune-related biological processes and key target genes of Chinese medicine were predicted by Coremine Medical database.Results A total of 852 differential gout genes were screened.GO enrichment analysis was mainly enriched in the leukocyte chemotaxis,interleukin-2 production,and regulation of vasculature development;KEGG pathway was mainly enriched in IL-17 signaling pathway,Chemokine signaling pathway,and TNF signaling pathway.Ten key genes,including TNF,IL-6,IL-1β,CXCL8,FOS,and VEGFA etc,were selected.The experimental validation showed that the protein expression levels of key genes were consistent with the expression trends of the chip data.Immune infiltration showed that monocytes,memory B cells,CD8+T-cells and IDC were closely associated with gout.Herbal predictions had identified Ginseng,Panax notoginseng,Scutellaria,Baicalensis,Saururus chinensis,and Salvia miltiorrhiza as potential drugs for the treatment of gout.Conclusion This study identified possible key genes and immune mechanisms contributing to the pathogenesis of gout,and discovered potential targetable traditional Chinese medicine.These key genes and herbs are expected to offer new insights and methods for the treatment of gout.

关 键 词：痛风 中药 生物信息学 免疫标志物 关节炎 代谢综合征 单钠尿酸盐 基因芯片 

分 类 号：R285[医药卫生—中药学]