基于生物信息学对骨关节炎铁超载关键基因的筛选与验证  

Screening and validation of Hub genes for iron overload in osteoarthritis based on bioinformatics

作　　者：邓柯淇 李光第[1,2] GOSWAMI ASHUTOSH 刘星余 何孝勇 Deng Keqi;Li Guangdi;Goswami Ashutosh;Liu Xingyu;He Xiaoyong(Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Department of Orthopedics,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学,贵州省贵阳市550004 [2]贵州医科大学附属医院骨外科,贵州省贵阳市550004

出　　处：《中国组织工程研究》2025年第9期1972-1980,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区科学基金项目(82160432),项目负责人:李光第;贵州省科技厅基础研究项目(黔科合基础-ZK[2022]-一般427),项目负责人:李光第;贵州医科大学国家自然科学基金培育项目(20NSP045),项目负责人:李光第。

摘　　要：背景:铁超载是指体内铁积累过多的情况,可引起各种组织的病理改变。目前,对于骨关节炎中铁超载相关的分子机制和潜在基因靶点,仍有待进一步的研究和探索。目的:通过生物信息学手段分析骨关节炎铁超载关键基因,并利用动物实验加以验证,以期为铁超载角度防治骨关节炎提供新的思路。方法:使用GEO数据库和GeneCards数据库筛选出与骨关节炎相关的基因和铁超载相关的基因。然后,取两者的交集,得到骨关节炎和铁超载共同相关的基因集合。采用GO和KEGG富集分析筛选这些基因的功能和通路。为了进一步研究这些基因之间的相互作用,构建PPI网络,并利用Cytoscape软件的5种计算方法来识别骨关节炎铁超载的关键基因(Hub基因)。将12只雄性SD大鼠分为骨关节炎组与对照组(正常大鼠),每组6只,骨关节炎组大鼠采用改良Hulth方法建立膝骨关节炎模型,并使用PCR检测Hub基因在两组大鼠膝关节组织中的表达情况。结果与结论:①共获得51个骨关节炎铁超载基因,GO富集分析表明其主要参与细胞因子受体结合、趋化因子受体结合、细胞因子活性、生长因子受体结合及寡糖结合等过程;②KEGG富集分析表明骨关节炎铁超载基因主要参与肿瘤坏死因子信号通路、脂质和动脉粥样硬化等信号通路;③构建蛋白质-蛋白质相互作用网络,进一步分析获得细胞间黏附分子1(intercellular cell adhesion molecule-1,ICAM-1)、肿瘤坏死因子配体超家族成员11(tumor necrosis factor superfamily member 11,TNFSF11)、骨髓瘤细胞癌基因(myelocytomatosis oncogene,MYC)、Janus激酶2(janus kinase 2,JAK2)及白细胞介素6,5个骨关节炎铁超载Hub基因,动物实验证实其在对照组与骨关节炎组大鼠膝关节组织中的表达有显著差异(P<0.05);④结果显示,ICAM-1,TNFSF11,MYC,JAK2及白细胞介素6可作为骨关节炎铁超载的关键基因,有望成为防治骨关节炎的新靶�BACKGROUND:Iron overload refers to excessive accumulation of iron in the body,which can cause pathological changes in various tissues.At present,the molecular mechanism and potential gene targets related to iron overload in osteoarthritis still need to be further studied and explored.OBJECTIVE:To analyze the key genes of iron overload in osteoarthritis by means of bioinformatics and verify them in animal experiments,so as to provide a new idea for preventing and treating osteoarthritis from the perspective of iron overload.METHODS:GEO database and GeneCards database were used to screen out genes associated with osteoarthritis and genes associated with iron overload.Then,the intersection of the two was taken to obtain a collection of genes commonly associated with osteoarthritis and iron overload.GO and KEGG enrichment analyses were used to screen the functions and pathways of these genes.To further investigate the interactions between these genes,a protein-protein interaction network was constructed and five computational methods of Cytoscape software were utilized to identify the Hub genes for iron overload in osteoarthritis.Finally,12 male Sprague-Dawley rats were divided into osteoarthritis group and normal group,with 6 rats in each group.A knee osteoarthritis model was established by the modified Hulth method in the osteoarthritis group.The expression of Hub genes in the knee joint of rats in each group was detected by PCR.RESULTS AND CONCLUSION:(1)A total of 51 genes associated with iron overload were identified in osteoarthritis.GO enrichment analysis showed that these genes were mainly involved in cytokine receptor binding,chemokine receptor binding,cytokine activity,growth factor receptor binding and oligosaccharide binding.(2)KEGG enrichment analysis showed that genes associated with iron overload in osteoarthritis was mainly involved in tumor necrosis factor signaling pathway and lipid and atherosclerosis signaling pathway.(3)The protein-protein interaction network was constructed,and five Hub genes of

关 键 词：骨关节炎 铁超载 关键基因 差异基因 生物信息学 动物实验 实验验证 生物标志物 

分 类 号：R459.9[医药卫生—治疗学] R363[医药卫生—临床医学] R364