基于人TNF-α转基因关节炎小鼠建立寒痹模型及评价  

Establishment and evaluation of cold arthralgia model based on human TNF-α transgenic arthritis mice

作　　者：朱兴旺 赵文婷 赵晓峰 毕亚男 左芳芳 陈恩生(指导) 肖长虹(指导) ZHU Xingwang;ZHAO Wenting;ZHAO Xiaofeng;BI Yanan;ZUO Fangfang;CHEN Ensheng;XIAO Changhong(Integrated Hospital of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510315,China)

机构地区：[1]南方医科大学中西医结合医院,广州510315

出　　处：《中国免疫学杂志》2022年第4期448-452,461,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81673723);国家自然科学青年基金项目(81804051);广东省中医药局科研课题项目(20191232);全国中医临床特色技术传承骨干人才培训项目[国中医药人教函(2019)36号]。

摘　　要：目的:采用人工气候环境干预人TNF-α转基因关节炎(Tgtc)小鼠构建寒痹模型,并初步探讨其发病机制及客观评价指标。方法:27只SPF级雌性Tgtc小鼠、27只SPF级雌性FVB小鼠随机分为空白对照组(BC)、风寒湿组(FHS)、风湿热组(FSR),每组18只,BC组置于SPF环境,FHS组接受人工FHS环境干预,FSR组接受人工FSR环境干预,2次/d,2 h/次,连续干预14 d。测量各组小鼠踝关节直径、四肢关节变形指数、踝关节皮肤颜色、踝关节体表血流灌注量,观察至第28天进行血清缺氧诱导因子1α(HIF1α)与热休克蛋白70(HSP70)、踝关节组织病理及免疫组化检测。结果:FVB小鼠观察测量指标不受FHS或FRS环境因素影响。Tgtc小鼠在人工气候干预期间,FHS组踝关节处皮肤颜色暗红程度、踝关节直径、四肢关节变形指数较BC组升高(P<0.05),而FSR组较BC组降低(P<0.05),环境干预结束后,3组Tgtc小鼠踝关节皮肤颜色、踝关节直径、关节变形指数差异无统计学意义(P>0.05);与BC组相比,FHS组小鼠踝关节及掌指关节处体表血流灌注量降低(P<0.05),滑膜组织HIF1α表达升高(P<0.05),而血清HIF1α含量、滑膜组织HSP70表达差异无统计学意义(P>0.05);FSR组掌指关节处体表血流灌注量、滑膜组织及血清HSP70表达、血清HIF1α含量均较BC组升高(P<0.05),滑膜组织中HIF1α表达较BC组降低(P<0.05);踝关节HE染色结果显示3组小鼠均有严重的滑膜成纤维细胞增生,伴有少量炎症细胞浸润,但FSR组软骨层损伤深度、软骨表面损伤范围、软骨处炎症细胞浸润程度较BC组降低。结论:Tgtc小鼠基础属性为寒,持续人工FHS环境干预时可作为寒痹模型,其发病与滑膜组织HIF1α上调有关;寒痹动物模型病理实质为滑膜成纤维细胞增生,伴少量炎症细胞浸润。皮肤颜色、关节体表血流灌注量可作为模型评估的客观量化指标。Objective:To establish a cold arthralgia model by treating human TNF-α transgenic mice in artificial climate chamber and to explore pathogenesis of cold arthralgia model and its objective evaluation index.Methods:27 SPF female Tgtc mice and 27 SPF female FVB mice were randomly divided into blank control group(BC),wind cold dampness group(FHS)and rheumatic fever group(FSR),with 18 mice in each group. Mice in BC group were placed in SPF environment,FHS group in artificial FHS environment and FSR group in artificial FSR environment,2 times/d,2 h/time,for continuous 14 d. Ankle joint diameter,limb joint deformation index,skin color of ankle joint and laser blood flow imaging were measured,serum levels hypoxia inducible factor 1(HIF1α)and heat shock protein 70(HSP70)expressions,ankle joint histopathology and immunohistochemistry were detected at 28 d.Results:FVB mice were not affected after exposure in FHS or FRS environment. Skin color of ankle joint,ankle joint diameter and limb joint deformation index of Tgtc mice in FHS group were higher than those in BC group(P<0.05),while these indexes in FSR group were lower than BC group(P<0.05). Skin color,ankle diameter and joint deformation index of Tgtc mice in three groups had no statistically difference after environmental intervention(P>0.05). Compared with BC group,SFI of ankle joint and metacarpophalangeal joint in FHS group was decreased(P<0.05),while expression of HIF1α in synovial tissue was increased(P<0.05). Levels of HIF1α in serum and HSP70 in synovial tissue had no statistically difference(P>0.05). SFI value,HSP70 in synovial tissue and serum,and HIF1α in serum of FSR group were increased(P<0.05),while HIF1α in synovial tissue was decreased(P<0.05). HE staining showed synovial fibroblast proliferation in three groups,with a small amount of inflammatory cell infiltration. However,cartilage injury and inflammatory cell infiltration in cartilage were alleviated in FSR group.Conclusion:Basic property of Tgtc mice is cold,Tgtc mice in artificial FHS environme

关 键 词：寒痹 Tgtc小鼠 人工气候环境 HIF1Α HSP70 模型评价指标 

分 类 号：R259[医药卫生—中西医结合] R332[医药卫生—中医内科学]