类风湿关节炎肾虚证大鼠模型的建立  

Establishment of a rat model of rheumatoid arthritis with kidney deficiency syndrome

作　　者：王燕[1] 赵宏艳[2] 刘梅洁[2] 肖诚[3] 吕诚[4] 谭勇[4] 査青林 李艳[2] 刘红[2] 吕爱平[4] 鞠大宏[2] 

机构地区：[1]民航总医院病理科,北京100123 [2]中国中医科学院中医基础理论研究所,北京100700 [3]中日友好医院临床医学研究所,北京100029 [4]中国中医科学院中医临床基础医学研究所,北京100700 [5]江西中医学院数据分析中心,江西南昌330006

出　　处：《中西医结合学报》2011年第9期973-982,共10页Journal of Chinese Integrative Medicine

基　　金："重大新药创制"科技重大专项资助项目(No.2009ZX09502-019);国家科技支撑计划重点课题资助项目(No.2006BAI08B01-01);国家自然科学基金资助项目(No.30701067;No.30902000)

摘　　要：目的:建立类风湿关节炎(rheumatoid arthritis,RA)肾虚证病证结合大鼠模型。方法:选用6周龄无特殊病原体级Sprague-Dawley大鼠110只,雌雄各半。分别采用去势法和羟基脲法制作肾虚动物模型,然后采用尾根部皮下注射Ⅱ型胶原与不完全弗氏佐剂混合物的方法诱导出关节炎。通过对大鼠一般状态观察,关节炎指数计算和关节肿胀度测量对比两种RA肾虚证病证结合动物模型的发病情况,并采用酶联免疫吸附测定法检测血浆抗Ⅱ型胶原抗体、白细胞介素6(interleukin-6,IL-6)、IL-10、γ干扰素(interferon-γ,IFN-γ)和皮质酮(corticosterone,CORT)含量,放射免疫法检测肾上腺环腺苷酸(cyclic adenylic acid,cAMP)和环鸟苷酸(cyclic guanylic acid,cGMP)含量。结果:与胶原诱导性关节炎(collagen-induced arthritis,CIA)对照组大鼠相比,结合去势法、羟基脲法制作的肾虚CIA大鼠关节红肿程度和关节损伤程度明显加重,并且这两组大鼠出现倦怠、精神萎靡,弓背蜷缩,体毛枯疏,自主活动少,反应逐渐迟钝,喜扎堆,饮食减少,大便质稀,肛周污秽等与临床肾虚证相似的症状;肾虚CIA大鼠cAMP与cGMP比值明显降低,雄性大鼠CORT含量升高,雌性大鼠的CORT水平下降,抗Ⅱ型胶原抗体含量明显升高。采用去势法制作的肾虚CIA大鼠与CIA对照组大鼠相比,IL-6、IL-10及IFN-γ水平明显升高。结论:用去势法和羟基脲法制作的肾虚证RA动物模型,既有明显的肾虚症状,又有明显的RA特征,说明这两种肾虚型RA病证结合模型能够部分地反映肾虚证RA病人的发病特点,具有可行性。而去势法制作的肾虚CIA大鼠更适合RA肾虚证的实验研究。Objective： To establish a rat model of rheumatoid arthritis （RA） with kidney deficiency syndrome.Methods： A total of 110 six-week-old specific pathogen-free male and female Sprague-Dawley rats were randomly divided into normal control group, sham-operated group, collagen-induced arthritis （CIA） control group, castration plus CIA group and hydroxyurea plus CIA group. Testiculus or ovary of rats in the castration plus CIA group was cut off, respectively. Rats in the hydroxyurea plus CIA group were given 375 mg/（kg·d） hydroxyurea by gavage administration for 17 d. Then rats in the CIA control group, castration plus CIA group and hydroxyurea plus CIA group were subcutaneously injected with mixture of type Ⅱ collagen and incomplete Freund＇s adjuvant to induce rheumatoid arthritis. General state, arthritis index and joint swelling of the rats were observed to evaluate the onset of CIA. Contents of anti-type Ⅱ collagen antibody, interleukin-6 （IL-6）, IL-10, interferon-γ （IFN-γ） and corticosterone （CORT） in plasma were detected by enzyme-linked immunosorbent assay, and adrenal cyclic adenylic acid （cAMP） and cyclic guanylic acid （cGMP） levels were detected by radioimmunoassay.Results： Compared with the CIA control group, the degrees of joint swelling and joint damage were significantly increased in the kidney-deficiency CIA rats （castration plus CIA group and hydroxyurea plus CIA group）, with kidney deficiency syndrome similar to human clinical symptoms, such as depressed, bowed back, dullness, reduced diet and perianal contamination; the rats in those two groups were noted with a significantly decreased ratio of cAMP/cGMP; the content of CORT was increased in male rats while decreased in female rats, with an obvious increase in the content of anti-type Ⅱ collagen antibody; the contents of IFN-γ, IL-6 and IL-10 were obviously increased in the castration plus CIA group. Conclusion： The rat model of RA with kidney deficiency syndrome has both obvious kidney deficiency

关 键 词：类风湿关节炎 病证结合动物模型 肾虚证 大鼠 

分 类 号：R-332[医药卫生]