四种不同抗原诱导的类风湿关节炎小鼠模型的病变特点比较研究  

Comparative study on pathological characteristics of four different antigen-induced rheumatoid arthritis mouse models

作　　者：杨超 胡智星 高双荣[1] 鄢泽然[2] 方罗昌婷 王潇潇 李群 陶庆文[2] 刘春芳[1] 林娜[1] YANG Chao;HU Zhi-xing;GAO Shuang-rong;YAN Ze-ran;FANG Luo-chang-ting;WANG Xiao-xiao;LI Qun;TAO Qing-wen;LIU Chun-fang;LIN Na(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;China-Japan Friendship Hospital,Beijing 100029,China)

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]中日友好医院,北京100029

出　　处：《药学学报》2023年第6期1586-1595,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上资助项目(81974529,82274176,81974526)。

摘　　要：类风湿关节炎(RA)是一种由抗原驱动和T细胞介导的自身免疫性疾病。二型胶原(CII)和纤维蛋白原(Fib)是RA发病进程中的两种主要抗原,其瓜氨酸化修饰(Cit)后产生的抗原也是诱导机体产生致病性抗瓜氨酸化蛋白抗体(ACPA)的诱因之一。为了给RA相关研究提供参考,本研究拟通过CII、Cit-CII、Fib和Cit-Fib 4种抗原分别与完全弗氏佐剂乳化,免疫DBA/1小鼠建立RA动物模型,从病理、影像和血清生化等方面对不同抗原所诱导模型的病变特点进行比较,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。结果显示, CII、Cit-CII和Cit-Fib诱导组小鼠均出现关节红肿、足趾变形等症状,临床积分和发病率均较正常组升高,其中CII组病变最严重,发病率达100%, Cit-CII和Cit-Fib组症状较轻,发病率分别为25%和37.5%;病理学和影像学检测结果发现CII诱导组小鼠关节出现严重的滑膜炎症、软骨和骨破坏, Cit-CII和Cit-Fib组仅出现轻微的炎症浸润、关节腔狭窄和骨破坏;血清抗体检测结果显示, CII、Cit-CII和Cit-Fib组均产生较高水平的抗环瓜氨酸肽(CCP)抗体,其中Cit-Fib组> Cit-CII组> CII组> Fib组,且Cit-CII和Cit-Fib组均产生高水平的瓜氨酸化表位特异性抗体,而总IgG水平则是CII组最高;血清ELISA和关节组织RT-PCR分析显示, CII诱导组的促炎因子和骨破坏相关分子表达量升高最明显,其次为Cit-Fib、Cit-CII。上述结果表明, 4种不同抗原中,由CII诱导的RA小鼠关节炎症状和病情最严重, IgG而非抗CCP抗体是其典型的免疫学特征, CII可作为RA小鼠造模首选;Cit-Fib具有一定的免疫原性,能部分诱导小鼠出现RA关节炎症状和病情,产生高水平抗CCP抗体和抗Cit-Fib抗体,较适合用于与瓜氨酸化相关的RA研究中;Cit-CII虽有一定的免疫原性,但其诱导的小鼠RA关节炎发生率和病情严重度较低。Rheumatoid arthritis(RA)is an autoimmune disease driven by antigens and mediated by T cells.Collagen II(CII)and fibrinogen(Fib)are the two main antigens in the pathogenesis of RA.The antigen produced after citrulline modification(Cit)is also one of the inducements to induce the body to produce a pathogenic anti-citrulline protein antibody(ACPA).To provide a reference for RA-related research,this study intends to establish an RA animal model by using CII,Cit-CII,Fib,and Cit-Fib antigens,emulsification with complete Freund's adjuvant and immunization with DBA/1 mice,respectively,to compare the pathological characteristics of RA models induced by different antigens from the aspects of pathology,imaging and serum biochemistry.Animal welfare and experimental process are in accordance with the regulations of the Experimental Animal Ethics Committee of the China Academy of Chinese Medical Sciences.The results showed that the CII,Cit-CII,and Cit-Fib induced mice all had symptoms such as joint redness and swelling,and toe deformation and the clinical score and incidence rate were higher than those of the normal group.The CII group had the most serious lesions,with a incidence rate of 100%,and the Cit-CII and Cit-Fib groups had mild symptoms,with a incidence rate of 25%and 37.5%,respectively;pathological and imaging examination results showed that the joints of mice in CII-induced group showed severe synovial inflammation,cartilage and bone destruction,while those in Cit-CII and Cit-Fib group showed only slight inflammatory infiltration,joint cavity stenosis and bone destruction;the results of serum antibody detection showed that CII,Cit-CII and Cit-Fib groups all produced high levels of anti-cyclic citrullinated peptide(CCP)antibodies,among which,Cit-Fib group>Cit-CII group>CII group>Fib group,and both Cit-CII and Cit-Fib groups produced high levels of citrullinated epitope-specific antibodies,while the total IgG level was the highest in CII group;serum ELISA and RT-PCR analysis of joint tissue showed that the expression

关 键 词：抗原 类风湿关节炎 免疫 病变特点 自身抗体 

分 类 号：R967[医药卫生—药理学]