基于生物信息学验证CCR5参与类风湿关节炎发生发展的遗传学和组织学证据  

Involvement of CCR5 in the development and progression of rheumatoid arthritis:genetic and histological evidence based on bioinformatics

作　　者：王友强 兰由玉 李世勇 WANG Youqiang;LAN Youyu;LI Shiyong(Department of Laboratory Medicine,the T.C.M Affiliated Hospital;Department of Rheumatology and Immunolo-gy of the Affiliated Hospital,Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属中医医院检验科,四川泸州646000 [2]西南医科大学附属医院风湿免疫科,四川泸州646000

出　　处：《西南医科大学学报》2021年第1期16-21,共6页Journal of Southwest Medical University

基　　金：西南医科大学校级课题(2019ZQN038)。

摘　　要：目的:通过生物信息学和实验方法探寻CC趋化因子受体5(CCR5)在类风湿关节炎(RA)的表达情况,以此验证CCR5参与RA发生发展的遗传学和组织学证据,并为后续利用RA大鼠模型研究CCR5的作用机制奠定基础。方法:在Gene Expression Omnibus(GEO)数据库中以“rheumatoid arthritis”为关键词检索RA的基因芯片GSE 97779、GSE 77298、GSE 10500、GSE 55457和GSE 1919,利用生物信息学分析工具GEO2R和limma包筛选差异基因以了解CCR5的表达情况,再通过免疫组化、qRT-PCR和Western Blot验证CCR5在RA大鼠中的异常表达情况。结果:在GSE 1919、GSE 97779、GSE 77298、GSE 10500、GSE 55457芯片中,CCR5在RA患者呈异常高表达,且CCR5是唯一在5个芯片均差异表达的基因。给大鼠注射弗氏完全佐剂10 d后,RA组大鼠关节炎指数(AI)明显升高,且随时间增加逐渐上升,至25 d时AI(7.12±1.13)较10 d(1.21±0.73)、15 d(4.87±1.21)时进一步增高(P<0.05);HE染色提示,RA组滑膜组织有较多炎性细胞浸润,可见新生血管形成,由此可见RA大鼠模型构建成功。免疫组化、qRT-PCR和Western Blot均提示RA组滑膜组织CCR5的表达水平较control组明显升高,与生信分析结果一致。结论:CCR5在RA患者及RA大鼠中均呈高表达,利用生物信息学和实验方法验证CCR5参与RA发生发展的遗传学和组织学证据是可行的,也为后续利用RA大鼠模型研究CCR5的具体作用机制奠定了理论基础。Objective:To explore the expression of C-C chemokine receptor 5(CCR5)in rheumatoid arthritis(RA)by bioinformatic and experimental approaches,to verify the genetic and histological evidence for the involve⁃ment of CCR5 in the development and progression of RA,and to provide a basis for the follow-up study of the mech⁃anism of action of CCR5 using a rat model of RA.Methods:The gene chips GSE 97779,GSE 77298,GSE 10500,GSE 55457,and GSE 1919 of RA were searched for in the Gene Expression Omnibus database using rheumatoid ar⁃thritis as a keyword.Differentially expressed genes were screened for using the bioinformatics analysis tools,GEO2R and limma,to understand CCR5 expression.The abnormal expression of CCR5 in RA rats was evaluated by immunohistochemistry,qRT-PCR,and Western blot.Results:In the GSE 1919,GSE 97779,GSE 77298,GSE 10500,and GSE 55457 chips,CCR5 expression was abnormally high in RA patients.Moreover,CCR5 was the on⁃ly gene that was differentially expressed on the five chips.At 10 days after injection of Freund’s complete adjuvant,the rats in the RA group had a substantially increased arthritis index(AI),which gradually increased with time;AI was significantly higher at 25 days than at 10 and 15 days(7.12±1.13 vs 1.21±0.73 vs 4.87±1.21,P<0.05).The result of HE staining showed neovascularization and inflammatory cell infiltration in the synovial tissue of the RA group,suggesting successful establishment of a rat model of RA.The results of immunohistochemistry,qRT-PCR,and Western blot all indicated that the expression level of CCR5 in synovial tissue was substantially higher in the RA group than in the control group,which was consistent with the results of bioinformatics analysis.Conclusion:CCR5 expression is high in RA patients and RA rats.It is feasible to use bioinformatic and experimental approaches to verify the genetic and histological evidence for the involvement of CCR5 in the development and progression of RA.This study provides a theoretical basis for the follow-up study of the mechan

关 键 词：类风湿关节炎 CCR5 生物信息学 

分 类 号：R593[医药卫生—内科学]