肾上腺素受体自身抗体通过上调趋化因子CXCL16诱导心功能不全  

Autoantibodies against the second extracellular loop of the β1-adrenoceptor induced cardiac insuffi-ciency through up-regulating chemokine CXCL16

作　　者：张诗晗 杜芸辉[2] 于海存[1] 何立娟 闫莉[4] 李玉明[5] Zhang Shihan;Du Yunhui;Yu Haicun;He Lijuan;Yan Li;Li Yuming(Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;Beijing Institute of Heart, Lung, and Blood Vessel Diseases, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China;Department of Laboratory Medicine, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, Chin;Institute of Basic Medical Sciences, Peking Union Medical College, Beijing 100005, Chin;Department of Basic Medical Sciences, Yanfing Medical College, Capital Medical University, Beijing 101300, Chin)

机构地区：[1]首都医科大学基础医学院生理学与病理生理学系,100069 [2]首都医科大学附属安贞医院北京心肺血管疾病研究所,100029 [3]首都医科大学附属安贞医院检验科,100029 [4]北京协和医学院基础研究所,100005 [5]首都医科大学燕京医学院,101300

出　　处：《中华微生物学和免疫学杂志》2018年第2期130-138,共9页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（81271165,81470540）;北京市自然科学基金（7142019）

摘　　要：目的 利用标记法芯片技术筛选β1肾上腺素受体自身抗体（β1-AA）致心功能不全的细胞因子,并对其中参与的信号通路进行分析.方法 收集67例冠心病患者和42例健康体检者血清,ELISA法检测血清中β1-AA水平;建立β1-AA被动免疫小鼠模型,采用小动物超声动态监测小鼠心脏功能及结构变化;苏木精-伊红（HE）及马松（Masson）染色观察小鼠心肌组织形态改变;标记法芯片技术筛选被动免疫模型鼠外周血清中β1-AA致心肌损伤的相关细胞因子;GO分析和KEGG通路富集分析对差异表达的细胞因子进行功能分类;ELISA法检测冠心病患者血清中该细胞因子水平,并进一步分析其与β1-AA之间的相关性.结果 冠心病患者血清中β1-AA滴度及阳性率明显高于对照组（P〈0.001）;β1-AA被动免疫8周诱导小鼠心功能不全;标记法芯片技术筛选出37个差异表达的细胞因子,其中趋化因子CXCL16的表达明显高于对照组（改变倍数〉20）;GO分析显示该细胞因子主要参与正向调节细胞死亡、迁移、运转及细胞组分改变等重要的生命过程,KEGG通路富集显示CXCL16显著富集在细胞因子-细胞因子受体相互作用信号通路及趋化因子信号通路;ELISA结果显示,β1-AA阳性冠心病患者血清中CXCL16的水平显著高于β1-AA阴性组（P〈0.01）,且与β1-AA之间成正向相关（P〈0.01,r=0.43）.结论 趋化因子CXCL16可能是β1-AA诱导心功能不全的关键细胞因子.Objective To screen out the cytokines relating to cardiac insufficiency caused by au-toantibodies against the second extracellular loop of the β1-adrenoceptor（β1-AA） using cytokine chip tech-nique, and to analyze the changes in signaling pathways. Methods Blood samples were collected from 67 patients with coronary artery disease（CAD） and 42 healthy subjects. ELISA was performed to detect β1-AA in plasma. BALB/c mice were passively immunized with the monoclonal antibodies against β1-AA （β1-AA mAb）. Dynamic changes in mouse cardiac structure and functions were detected by heart ultrasound. Hema-toxylin and eosin （HE） and Masson staining were used to observe morphological changes in heart tissues.Cytokine chip technique was used to screen out the cytokines causing myocardial injury. Gene Ontology （GO） and KEGG （Kyoto Encyclopedia of Genes and Genomes） pathway enrichment analysis were used to classify the differentially expressed cytokines. Results Patients with CAD showed increased titer and posi-tive rate of β1-AA as compared with healthy subjects（P〈0.001）. The mouse model of heart injury was in-duced by β1-AA at the 8th week after immunization. A total of 37 differentially expressed cytokines were found in the model group,of which 11 cytokines were up-regulated and 26 cytokines were down-regulate as compared with those in the mouse control group. The level of CXCL16 was significantly increased in β1-AA-positive mice. GO analysis showed that CXCL16 was mainly involved in life processes including the positive regulation of cell death, migration, locomotion and cellular component movement. KEGG pathway enrich-ment analysis showed CXCL16 was significantly enriched in the pathway of cytokine-cytokine receptor inter-action and chemokine signaling pathway. ELISA showed that compared with β1-AA-negative patients, CXCL16 level was significantly increased in β1-AA-positive patients （P〈0.01）. A positive correlation was found between β1-AA and CXCL16 （P〈0.01,r=0.43）. Conclu

关 键 词：β1肾上腺素受体自身抗体 心功能不全 CXCL16 

分 类 号：R541.6[医药卫生—心血管疾病]