多组学分析揭示溃疡性结肠炎缓解期脾虚湿困证与活动期湿热阻滞证间的潜在分子机制及靶向中药预测  

Revelation of potential molecular mechanism between spleen deficiency and dampness-stagnation syndrome in remission stage and damp-heat stagnation syndrome in active stage of ulcerative colitis based on multiomics analysis and prediction of targeted traditional Chinese medicine

作　　者：黎祖鸣 封杰妮 陈雪如 陈剑坤 卢月 李际强 冯艳 LI Zuming;FENG Jieni;CHEN Xueru;CHEN Jiankun;LU Yue;LI Jiqiang;FENG Yan(The Second Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The Second Affiliated Hospital of Guangzhou University of Chinese Medicine(Guangdong Hospital of Traditional Chinese Medicine),Guangzhou 510006,China)

机构地区：[1]广州中医药大学第二临床医学院,广东广州510405 [2]广州中医药大学第二附属医院(广东省中医院),广东广州510006

出　　处：《中草药》2024年第9期3041-3056,共16页Chinese Traditional and Herbal Drugs

基　　金：广州市科技计划⁃市校(院)联合资助基础与应用基础研究项目(2023A03J0738);广东省中医院朝阳人才科研专项资助(ZY2022KY10,ZY2022YL04)。

摘　　要：目的综合分析溃疡性结肠炎(ulcerative colitis,UC)相关scRNA-seq和RNA-seq数据集,探讨UC缓解期脾虚湿困证与活动期湿热阻滞证间的潜在分子机制,并挖掘潜在干预中药。方法运用差异基因表达分析和韦恩图鉴定出UC缓解期脾虚湿困证和活动期湿热阻滞证相关基因及两者间的对话基因。通过京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)、基因集变异分析(gene set variation analysis,GSVA)及Spearman相关性分析深入探讨对话基因参与的生物过程及潜在功能。通过单细胞转录组分析探讨UC缓解期与活动期的差异情况及对话基因在其中的作用。基于循环算法构建UC缓解期脾虚湿困证向活动期湿热阻滞证发生发展的风险预测模型。运用外部数据集、动物实验与实时定量聚合酶链反应(realtime quantitative polymerase chain reaction,RT-PCR)进一步验证对话基因的表达情况。借助SoFDA数据库构建“基因-中医症状/现代医学症状”网络;借助TCMIP和COREMINE数据库预测对话基因的潜在靶向中药。结果鉴定出31个UC缓解期脾虚湿困证相关基因,其主要富集在代谢相关途径;鉴定出160个UC活动期湿热阻滞证相关基因,其主要富集在炎症免疫相关途径。鉴定出22个UC缓解期脾虚湿困证与活动期湿热阻滞证之间的对话基因,上调对话基因与炎症免疫相关途径呈显著相关性,下调对话基因与代谢途径呈显著相关性。单细胞转录组分析显示,相比于正常对照及UC缓解期,UC活动期中B细胞占比上调;B细胞在UC活动期湿热阻滞证中作用贡献较大,而在UC缓解期脾虚湿困证中的作用贡献较小;UC活动期B细胞的受体信号通路活性明显高于UC缓解期;上调的对话基因CD55在B细胞中表达水平较高,CD55+B细胞与CD55−B细胞之间存在差异的细胞通讯及细胞代谢水平。6个风险预测基因(CD55、PCSK1、SERPING1、ACAT1、SLC26A2、HMGCS2)可以Objective To explore the potential molecular mechanism between spleen deficiency and dampness-stagnation syndrome in remission stage and dampness-heat stagnation syndrome in active stage of ulcerative colitis(UC)by comprehensively analyzing the scRNA-seq and RNA-seq data sets related to UC,and to explore the potential intervention of traditional Chinese medicine(TCM).Methods Differential gene expression analysis and venn diagram analysis were used to identify the related genes of spleen deficiency and dampness-stagnation syndrome in remission stage of UC,dampness-heat stagnation syndrome in active stage of UC and the crosstalk genes between them.Enrichment analysis of Kyoto encyclopedia of genes and genomes(KEGG)and gene set variation analysis(GSVA)and Spearman correlation analysis were used to explore the biological processes involved in crosstalk genes and their potential functions.Based on single-cell transcriptome analysis,we explored the differences between remission and active phases of UC and the role of crosstalk genes in them.Based on the circulation algorithm,a risk prediction model for the occurrence and development of UC from spleen deficiency and dampness-stagnation syndrome in remission stage to dampness-heat stagnation syndrome in active stage was constructed.Extraneous data,animal experiments and real-time quantitative polymerase chain reaction(RT-PCR)were used to further verify the expression of crosstalk genes.Based on SoFDA database,we constructed“gene-TCM symptom/modern medical symptom”network.Potential target traditional Chinese medicines(TCMs)of crosstalk genes were predicted by TCMIP and COREMINE databases.Results A total of 31 genes related to spleen deficiency and dampness-stagnation syndrome in remission stage of UC were identified,which were mainly enriched in metabolism-related pathways;A total of 160 genes related to dampness-heat stagnation syndrome in active stage of UC were identified,which were mainly enriched in inflammatory immune-related pathways.A total of 22 crosstalk gen

关 键 词：溃疡性结肠炎 单细胞转录组 转录组 缓解期 脾虚湿困证 活动期 湿热阻滞证 人参 黄芪 枸杞子 

分 类 号：R285[医药卫生—中药学]