人类风湿性关节炎滑膜-软骨-NOD.scid小鼠嵌合体模型的建立

Chimera models of synovium-cartilage-NOD.scid mice with human rheumatoid arthritis

目的:建立能反应自身免疫特点的类风湿性关节炎动物模型,为进一步研究该病发病机制奠定基础。方法:实验于2006-03/12在南方医科大学实验动物中心完成。①实验材料:SPF级2～4个月龄NOD.scid小鼠20只,体质量17～22g,雌雄各半;人类风湿性关节炎滑膜组织(佛山市第一人民医院风湿科提供,患者知情同意);骨关节炎滑膜及正常软骨(南方医院创伤骨科提供,患者及供者知情同意)。②实验分组:将小鼠随机分为2组,类风湿性关节炎滑膜组和骨关节炎滑膜组,雌雄不限,每组10只。③实验过程:每组小鼠背部皮下植入的正常软骨组织,随后将类风湿性关节炎滑膜和骨关节炎滑膜植入软骨之上。④造模10周后麻醉下处死小鼠,用放射免疫法检测血清中肿瘤坏死因子α含量,移植物进行组织病理学观察,并进行组织学积分。结果:①模型鼠一般情况:实验期间NOD.scid小鼠无手摸皮骨感、活动度差、毛稀疏等典型移植物抗宿主疾病表现,模型鼠在SPF环境下10周存活率100%。②肿瘤坏死因子α含量:类风湿性关节炎滑膜组鼠血清中肿瘤坏死因子α放射含量类风湿性关节炎组明显高于骨关节炎组[(0.80±0.06),(0.70±0.03)μg/L,t=4.466,P<0.001]。③组织病理学观察:苏木精-伊红染色,骨关节炎滑膜组只见少量的滑膜细胞增生和炎症细胞,移植的滑膜组织主要被纤维组织形成的条索状物代替,无明显地软骨被侵蚀破坏发生;类风湿性关节炎滑膜组可明显见到大量的滑膜细胞增生和生化中心形成,病变部位的组织结构间质变为疏松,变为境界不清晰的颗粒状或块状无结构强嗜酸性红染物质;软骨边缘被滑膜组织侵蚀破坏明显;类风湿性关节炎滑膜组滑膜增生、软骨侵蚀和软骨降解积分均高于骨关节炎滑膜组(P<0.04)。结论:在NOD.scid体内可成功建立类风湿性关节炎动物模型。

AIM： To establish autoimmune model mice of rheumatoid arthritis for investigating the pathogenesis of rheumatoid arthritis. METHODS： The experiment was conducted in the Laboratory Animal Center of Southern Medical University from March to December 2006. ①Totally 20 SPF-level NOD.scid mice aged 2-4 months and with the body mass of 17-22 g of either sex （half and half） were selected. Human rheumatoid arthritis synovium tissues were collected from patients of Department of Rheumatoid Diseases, Foshan First People＇s Hospital with the agreement of patients. Ostearthritis synovium and normal human cartilage were collected from patients and donors of Department of Orthopedics and Trauma of Nanfang Hospital with the agreement of patients and donors. ②The mice were randomly divided into 2 groups, rheumatoid arthritis synovium group and ostearthritis synovium group with 10 in each group. ③Normal human cartilage was embedded in the back subcutaneously of mice in the two groups, then fresh synovial tissues derived from patients with rheumatoid arthritis and ostearthritis were implanted on the cartilage respectively. ④Mice were killed after 10 weeks under anesthesia. Tumor necrosis factor （TNF-α） content in serum was determined by radioimmunoassay （RIA）. The implanted tissues were taken for histopathological test and histological integral. RESULTS：①There should be no typical graft-versus-host disease （GVHD） behavior such as hand touching skin bone sense, bad mobility and sparse capill for NOD.scid mice during the experiment. Survival rate for 10 weeks of model mice under SPF environment was 100%,②The serum TNF-α level was obviously higher in mice of the rheumatoid arthritis synovium group than the ostearthritis synovium group [（0.80±0.06）, （0.70±0.03） μg/L,t =4.466,P 〈 0.001]. ③Haematoxylin-eosin staining showed that there was only a little amount of synovium cell proliferation and inflammatory cells in the ostearthritis synovium group. Implanted synovium tissues were mainly replaced by strip material formed from flbroplasia, without manifest cbrrosion and injury of cartilage. A great amount of synovial cell hyperplastic and biochemical center could be obviously seen in rheumatoid arthritis synovium group. Histological interstitial substance in disease part became loosen. It became granulation with unsharp boundary or block strong acidophic red dye substance without structure. Edge of cartilage was obviously damaged by corrosion of synovium tissue. Integrals of synovium proliferation, erosion of cartilage and degradation of cartilage were higher in the rheumatoid arthritis synovJum group than the ostearthritis synovium group （P 〈 0.04）, CONCLUSION; Model mice of rheumatoid arthritis can be successfully established in NOD.Scid mice.