基于症状映射的类风湿关节炎瘀血阻络证生物内涵研究及其病证结合动物模型建立和评价  

Exploring biological connotation of blood stasis syndrome of rheumatoid arthritis and establishment of improved animal models based on syndrome-symptom mapping

作　　者：陈文佳 李涛 胥明珠 巩勋[3] 刘蔚翔 李培豪 姜泉[3] 刘维[4] 毛霞[1] 李欣[1] 许海玉[1] 林娜[1] 张彦琼 CHEN Wen-jia;LI Tao;XU Ming-zhu;GONG Xun;LIU Wei-xiang;LI Pei-hao;JIANG Quan;LIU Wei;MAO Xia;LI Xin;XU Hai-yu;LIN Na;ZHANG Yan-qiong(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Chinese Materia Medica,Yunnan University of Chinese Medicine,Kunming 650500,China;Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China;First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China)

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]云南中医药大学中药学院,云南昆明650500 [3]中国中医科学院广安门医院,北京100053 [4]天津中医药大学第一附属医院,天津300193

出　　处：《药学学报》2023年第8期2434-2441,共8页Acta Pharmaceutica Sinica

基　　金：国家重点研发计划项目(2018YFC1705201);中国中医科学院科技创新工程项目(CI2021A03807).

摘　　要：瘀血阻络证是类风湿关节炎(rheumatoid arthritis,RA)的临床核心证候之一,但其证候生物内涵尚不明确,同时也缺乏符合病证特点的病证结合动物模型。本研究旨在筛选RA瘀血阻络证的候选生物标志基因集,揭示其证候生物内涵,同时在此基础上对病证结合动物模型的制备方法从病、证、症特点进行探索和评价。本研究经中国中医科学院广安门医院(批准号:2019-073-KY-01)和天津中医药大学第一附属医院(批准号:TYLL2021[K]字018)伦理委员会批准,研究对象知情同意。动物福利和实验过程均遵循中国中医科学院实验动物伦理委员会的规定(批准号:IBTCMCACMS21-2207-01)。临床收集RA瘀血阻络证(3例)、非瘀血阻络证(7种证型,3例/证型)及健康志愿者(4例)的全血样本,开展转录组测序、KEGG分析、基因集富集分析(gene set enrichment analysis,GSEA)和加权基因共表达网络分析(weighted correlation network analysis,WGCNA),共筛选到126个枢纽基因,其功能注释结果显著富集于“免疫−炎症”相关通路和脂质代谢调节(鞘磷脂、醚脂质代谢和类固醇生物合成)相关通路。对枢纽基因集与中医主次症、西医表型症状和病理环节相关的靶标基因集做症状映射分析结果显示,关节刺痛、关节形态异常、瘀斑和血液循环异常是RA瘀血阻络证的代表性病证特点。病证结合动物模型实验结果表明,相比单纯胶原诱导(collagen-induced arthritis,CIA)模型组,CIA联合盐酸肾上腺素(adrenaline hydrochloride,Adr)3模型组大鼠加剧了血液流变性、凝血功能、血小板功能和内皮功能的异常变化,提示该种瘀血阻络证RA大鼠可能处于“血瘀”状态。本研究结果有助于推进RA瘀血阻络证的证候客观化研究,并为建立反映临床特点的病证结合动物模型提供新思路。Blood stasis syndrome is one of the core clinical syndrome of rheumatoid arthritis(RA),but the biological connotation of this syndrome is not clear,and there is a lack of disease improved animal models that match the characteristics of this disease and syndrome.The aim of this study was to screen the candidate biomarker gene set of blood stasis syndrome of RA,reveal the biological connotation of this syndrome,and explore and evaluate the preparation method of the improved animal model based on the characteristics of"disease-syndromesymptom".The study was approved by the ethics committee of Guang'anmen Hospital,Chinese Academy of Traditional Chinese Medicine(No.2019-073-KY-01)and the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine(No.TYLL2021[K]018),and the study subjects gave their informed consent.Animal welfare and experimental procedures followed the regulations of the Experimental Animal Ethics Committee of the Chinese Academy of Traditional Chinese Medicine(No.IBTCMCACMS21-2207-01).The whole blood samples were collected clinically from RA patients with blood stasis syndrome(3 cases)or other syndromes(7 types,3 cases/type),and healthy volunteers(4 cases),and then transcriptome sequencing,KEGG,gene set enrichment analysis(GSEA)and weighted correlation network analysis(WGCNA)analysis were performed.126 pivotal genes were screened,and their functional annotation results were significantly enriched in"immune-inflammation"related pathways and lipid metabolism regulation(sphingolipids,ether lipid metabolism and steroid biosynthesis).Syndrome-symptom mapping of hub gene set to the TCM primary and secondary symptoms,Western phenotypic symptoms and pathological links showed that joint tingling,abnormal joint morphology,petechiae and abnormal blood circulation are representative of blood stasis syndrome of RA.The results of the improved animal model showed that the rats in the collagen-induced arthritis+adrenaline hydrochloride(CIA+Adr)3 model group had increased blood rheology,coagulati

关 键 词：类风湿关节炎 瘀血阻络证 转录组学 加权基因共表达网络分析 症状映射 病证结合 

分 类 号：R967[医药卫生—药理学]