Page 43 - 2023-08-中国全科医学

·940· http: //www.chinagp.net E-mail: zgqkyx@chinagp.net.cn March 2023, Vol.26 No.8

1.Shanxi Medical University，Taiyuan 030000，China
2.Cardiovascular Department，Xinzhou People's Hospital，Xinzhou 034000，China
3.Cardiovascular Department，Second Hospital of Shanxi Medical University，Taiyuan 030000，China
*
Corresponding author：BIAN Yunfei，Professor，Doctoral supervisor；E-mail：sdeybyunfei@163.com
【Abstract】 Background Cardiovascular disease（CVD） is a common and frequently occurring disease，and
the prevalence and mortality of which are increasing rapidly. Atherosclerosis（AS） is the pathological basis of ischemic CVD.
Studies have shown that epicardial adipose tissue（EAT） promotes the progression of AS by secreting exosomes and bioactive
substances，but the mechanism of action still needs to be further studied. Objective To perform a bioinformatics analysis of role
of EAT in coronary artery disease（CAD）at cellular and molecular levels by identifying the differentially expressed genes（DEGs）
in EAT to explore the status of immune cell infiltration，and to assess and verify whether EAT is derived from DEGs in exosomes in
CAD patients. Methods We downloaded GSE64554 and GSE120774 datasets about EAT from the GEO database and performed
a bioinformatics analysis using R language and related packages. We first used R language to screen the DEGs in EAT and CAD
patients，then used GO/KEGG enrichment analysis to establish a protein interaction network to explore biological functions of the
screened genes and transcription factors potentially involved in their regulation process. After that，we conducted a weighted gene
co-expression network analysis（WGCNA）of EAT in GSE64554 dataset to obtain a gene module related to CAD phenotype，
then crossed the hub genes in this module and DEGs in EAT to obtain the key common genes. We used Cibersort to characterize
the immune cell infiltration in EAT. Then we obtained DEGs from blood exosomes of CAD patients and healthy controls included
in the exoRbase database，crossed DEGs in EAT and blood exosomes to identify the common genes to be used as diagnostic and
therapeutic markers for CAD，and their values were tested by qRT-PCR measurement of clinical samples. The selected genes
were analyzed by GO/KEGG and Metascape enrichment analyses. Results A total of 1 511 DEGs in EAT of CAD patients were
identified，including 956 with up-regulated expression and 555 with down-regulated expression. By crossing the DEGs in EAT
and hub genes in modules associated with CAD closely，we identified DDX47，FEM1C，NOL11，SRP54，ABI1，PATL1，
BNIP2，C1orf159，and CHCHD4 as key genes in the development of CAD. Immune cell infiltration analysis showed that the
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abundance of immature CD 4 T cells increased while expression abundance of resting dendritic cells decreased in EAT of CAD
patients（P<0.05）. A total of 1 658 DEGs in exosomes of CAD patients，including 278 with up-regulated expression and 1 380
with down-regulated expression. One hundred and twenty-nine common DEGs were obtained by cross-tabbing DEGs in EAT
and exosomes of CAD patients，among which BPI，BIRC5，CXCL12，RNASE1 and F2R with higher expression abundance
were selected as potential diagnostic and therapeutic markers for CAD. By qRT-PCR detection，CAD patients were found with
increased mRNA expression levels of BPI，BIRC5，CXCL12，RNASE1（P>0.05），and decreased F2RmRNA expression
level（P<0.05） than controls. GO/KEGG enrichment analysis showed that DEGs in EAT were mainly involved in the cytosol，
MHC protein complex，RNA degradation，antigen processing and presentation. A PPI network was built，in which RPS27A
gene was identified as a gene with the highest degree of connectivity by use of Cytoscape plugin CytoHubba with MCC algorithm.
Metascape enrich analysis indicated that DEGs enriched mainly in cellular response to DNA damage，RNA metabolism，
regulation of cell stress responses，and adaptive immune system. By an analysis of TRRUST datasets，we predicted that
transcription factor CIITA may play a role in the regulation of DEGs in EAT influencing CAD. Conclusion EAT may be involved
in the development of CAD through proinflammatory and immune pathways，in which DDX47，FEM1C，NOL11，SRP54，
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ABI1，PATL1，BNIP2，C1orf159，CHCHD4 and RPS27A may play a vital role as the key genes. The abundance of naive CD 4
T cells significantly increased while that of resting dendritic cells decreased obviously in EAT from CAD patients. BPI，BIRC5，
CXCL12，RNASE1 and F2R may be excreted by EAT and have the potential as markers in CAD diagnosis and treatment.
【Key words】 Coronary artery disease；Epicardial adipose tissue；Exosome；Bioinformatics；Immune cell
infiltration；Key genes；Hub genes

动脉粥样硬化（atherosclerosis，AS）导致的心血管 利用基因芯片技术对临床患者标本进行检测分析，筛选
疾病（cardiovascular disease，CVD）是全球死亡的主要 出一些具有价值的基因，进而深入研究冠状动脉粥样硬
原因，世界卫生组织预测 2030 年将有 2 360 万人死于 化性心脏病（coronary artery disease，CAD）的发病机制
CVD ［1］ 。AS 形成机制复杂，主要涉及血管内皮损伤、 具有重要的临床意义。肥胖是 CVD 的独立危险因素，
单核巨噬细胞黏附、脂质沉积等 ［2］ ，但这些因素如何 但肥胖的主要评价指标体质指数（BMI）与 AS 患者死
影响 AS 进程，其机制尚未完全清楚。因此找寻 AS 新 亡率呈“U”型关联，这种现象被称为肥胖悖论，因此
的诊断靶点，预防 AS 的发生，延缓其进展尤为重要。 脂肪组织不再被认为是单纯的“能量仓库”，而是一种