泛素特异性蛋白酶18促进ABCG1泛素化对动脉粥样硬化形成的影响  

Effect of ubiquitin-specific protease 18 promoting ubiquitination of ABCG1 on the formation of atherosclerosis

作　　者：安洋 王小丽 王杰[1] 戴佳琳[1] AN Yang;WANG Xiaoli;WANG Jie;DAI Jialin(School of Forensic Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学法医学院,贵州贵阳550004

出　　处：《贵州医科大学学报》2025年第2期167-179,共13页Journal of Guizhou Medical University

基　　金：国家自然科学基金项目(82060340)。

摘　　要：目的探讨泛素特异性蛋白酶18(ubiquitin-specific protease 18,USP18)在动脉粥样硬化(atherosclerosis,AS)形成中的作用及潜在的机制。方法从基因表达数据库(GEO)下载冠状动脉疾病(coronary atherosclerotic heart disease,CHD)基因表达数据集GSE129935,使用GEO2R筛选差异表达基因(differential expression genes,DEGs),并进一步对DEGs进行基因本体(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)富集分析;收集冠心病患者(CHD组)和非冠心病患者(Non-CHD组)的冠状动脉,利用免疫组化(ICH)染色和Western blot检测冠脉中USP18的表达水平;使用ApoE^(-/-)小鼠+高脂饲料构建AS模型,随机分为单纯高脂对照组(NC组)、腺病毒对照组(Ad-NC组)及USP18基因敲低组(Ad-USP18组);高脂饮食及相应处理16周后,取主动脉,使用苏木精和伊红(HE)染色和油红O染色检测斑块形成和脂质沉积情况;THP-1衍生的巨噬细胞作为体外研究模型,通过Western blot检测ATP结合盒转运蛋白G1((ATP binding cassette subfamily G member 1,ABCG1)表达及泛素化水平,双重免疫荧光染色检测USP18与ABCG1的共定位表达,油红O染色检测泡沫细胞形成情况。结果GSE129935数据集鉴定出285个DEGs(上调139个,下调146个),其中USP18表达上调;GO及KEGG分析表明,DEGs主要富集于炎症、抗原结合等生物学过程,涉及NF-κB信号通路、趋化因子信号通路、脂质和AS等;CHD患者冠状动脉中USP18的表达上调(P<0.05),且与AS病变程度呈负相关(r=-0.525,P=0.006);敲低USP18基因可增加ApoE^(-/-)小鼠主动脉斑块负荷和脂质沉积、并降低主动脉中ABCG1的蛋白表达(P<0.05);USP18能够与ABCG1共定位表达,敲低USP18基因能够下调巨噬细胞中ABCG1的蛋白表达并促进脂质积累及泡沫细胞形成(P<0.05),并增加ABCG1的泛素化水平,加快ABCG1的蛋白降解(P<0.05)。结论USP18能够靶向结合ABCG1,促进ABCG1的泛素化水平、阻碍泡Objective To investigate the role and potential mechanism of ubiquitin-specific protease 18(USP18)in the formation of atherosclerosis(AS).Methods The gene expression dataset GSE129935 for coronary heart disease(CHD)was obtained from the Gene Expression Omnibus(GEO)database.GEO2R was used to screen differentially expressed genes(DEGs),and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was performed on DEGs.The coronary arteries samples of CHD patients(CHD group)and non-CHD patients(Non-CHD group)were collected,and the expression level of USP18 in coronary arteries was examined by immunohistochemistry(ICH)staining and Western blot.ApoE^(-/-)mice fed with high-fat diet were used to develop AS model,which were randomly divided into 3 groups:high-fat control group(NC group),adenovirus control group(Ad-NC group),and USP18 gene knockdown group(Ad-USP18 group).After high-fat diet and corresponding treatment for 16 weeks,the aorta was taken.The plaque formation and lipid deposition were assessed by hematoxylin and eosin(HE)staining and oil red O staining.THP-1-derived macrophages were used as an in vitro research model.The expression of ATP-binding cassette transporter G1(ABCG1)and ubiquitination level were detected by Western blot.The co-localization of USP18 and ABCG1 was investigated using double immunofluorescence staining and oil red O staining was used to detect the formation of foam cells.Results A total of 285 DEGs,comprising 139 up-regulated and 146 down-regulated genes,were identified in the GSE129935 dataset,with USP18 expression being up-regulated.GO and KEGG analysis indicated that these DEGs predominantly participate in biological processes such as inflammation and antigen binding,involving NF-κB signaling pathway,chemokine signaling pathway,lipid metabolism,and AS.The expression of USP18 was significantly elevated in the coronary arteries of patients with CHD(P<0.05)and negatively correlated with the severity of AS lesions(r=-0.525,P=0.006).Knockdown of the USP18 gene

关 键 词：动脉粥样硬化 生物信息学 泛素特异性蛋白酶18 ATP结合盒转运蛋白G1 泡沫细胞 

分 类 号：R541.4[医药卫生—心血管疾病]