基于综合生物信息学和单细胞测序方法揭示红花-丹参治疗冠心病的生物学和免疫学机制  

Unraveling the biological and immunological mechanisms of safflower-danshen in the treatment of coronary atherosclerotic heart disease:a comprehensive bioinformatics and single-cell sequencing approach

作　　者：魏东升 刘孝生 李路珍 齐佳杰 王雨轩 张哲[1,2,3,4] Dongsheng Wei;Xiaosheng Liu;Luzhen Li;Jiajie Qi;Yuxuan Wang;Zhe Zhang(Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Innovation Engineering Technology Center of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Key Laboratory of the Theory and Application of Viscera in Chinese Medicine,Ministry of Education,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110033,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学中医药创新工程技术中心,辽宁沈阳110847 [3]辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁沈阳110847 [4]辽宁中医药大学附属医院,辽宁沈阳110033

出　　处：《Journal of Chinese Pharmaceutical Sciences》2023年第10期796-812,共17页中国药学（英文版）

基　　金：National Administration of Traditional Chinese Medicine Supporting Project for Young Qi Huang Scholars (Grant No. 20201A2180)。

摘　　要：本文基于生物信息学、网络药理学与单细胞测序探讨红花-丹参治疗冠心病(CAD)的机制。采用Comparative Toxicogenomics Database、Swiss target prediction、Binding Database Home、TargetNet四个数据库筛选出羟基红花黄色素、丹参酮、丹酚酸、丹参素的靶向基因。基于GEO数据库所筛选的3个CAD数据集,通过差异分析及加权基因共表达网络(WGCNA)筛选与CAD相关的基因。融合药物与疾病基因,通过cytoscape中mcode插件筛选出药物-疾病调控网络核心亚群。使用ssGSEA算法分析38种免疫细胞与mcode核心亚群基因的浸润情况。使用单细胞RNA测序明确mcode核心亚群所分布的细胞亚群。最后,分别对药物靶点基因、疾病基因以及mcode核心亚群基因进行GO、KEGG富集分析。结果发现,四个数据库共得到485个药物靶点基因。通过差异分析及WGCNA得到617个疾病基因。mcode插件获得1个药物-疾病的核心调控亚群,包括99个基因。ssGSEA算法结果表明TGFβ家族成员、趋化因子、白介素受体可能是红花-丹参治疗CAD核心调控的免疫细胞。单细胞RNA测序结果表明巨噬细胞与单核细胞可能是核心细胞亚群。GO、KEGG富集分析结果表明内膜结合的细胞器与细胞核可能是核心细胞成分。红花-丹参可能通过调控TGFβ家族成员、趋化因子、白介素受体、巨噬细胞、单核细胞、内膜结合的细胞器与细胞核发挥治疗CAD的作用。In the present study,we investigated the mechanism of safflower-danshen treatment for coronary artery disease(CAD)through bioinformatics,network pharmacology,and single-cell sequencing.To identify target genes for hydroxy safflower yellow pigment,tanshinone,danshinolic acid,and tanshinol,we employed various databases,including Comparative Toxicogenomics Database,Swiss target prediction,Binding Database Home,and TargetNet.By analyzing three CAD datasets obtained from the GEO database,we identified CAD-associated genes using differential analysis and a weighted gene coexpression network analysis(WGCNA).We integrated drug-disease genes and utilized the mcode plugin in Cytoscape to identify core subgroups in the drug-disease regulatory networks.The infiltration of 38 immune cells/functions was assessed using the ssGSEA algorithm.Additionally,single-cell RNA sequencing(SCS)was employed to examine the distribution of cellular subpopulations within the mcode regulatory network.Finally,we performed GO and KEGG enrichment analyses for drug target genes,disease genes,and mcode core subpopulation genes.Our results revealed 485 drug target genes obtained from four databases.Through differential analysis and WGCNA,we identified 617 disease genes associated with CAD.The mcode plugin analysis yielded a drug-disease core regulatory subpopulation consisting of 99 genes.The ssGSEA algorithm indicated that members of the TGFβ family,chemokines,and interleukin receptors might play key roles in the immune regulation of safflower-danshen treatment for CAD.Furthermore,SCS results suggested that macrophages and monocytes might be core cell subpopulations within this context.GO and KEGG enrichment analyses highlighted endosome-bound organelles and nuclei as potentially important cellular components.In conclusion,safflower-danshen treatment for CAD might exert its therapeutic effects by modulating TGFβfamily members,chemokines,interleukin receptors,macrophages,monocytes,endosome-bound organelles,and nuclei.

关 键 词：红花-丹参 冠状动脉粥样硬化性心脏病 生物信息学 单细胞测序 网络药理学 

分 类 号：R392-3[医药卫生—免疫学]