一种新型类风湿关节炎中医寒痹与热痹动物模型的研究

Cold-Bi and Heat-Bi are two basic syndrome of RA(Rheumatoid Arthritis). The hypothesis was innovatively suggested in this project that RASF(Rheumatoid Arthritis Synovial Fibroblasts) proliferation is the main pathology of Cold-Bi, and the inflammatory response based on proliferation is the main pathology of Heat-Bi. So that the RASF spontaneous hyperplasia model should be choose as the basic diseases model for TCM(Traditional Chinese Medicine) syndrome models. TgTC mice is one of the local foster human tumor necrosis factor transgenic mice(hTNF Tg), which get joint swelling and deformity spontaneously without acute inflammatory reaction, and accompany with SF proliferative highly without inflammatory cell infiltration in pathology. We intended to serve TgTC mouse as the base model, collagen type Ⅱ immune the mice to induce immune inflammatory response, then stimulated with “the artificial climate box”by wind coldness dampness or wind dampness hotness . Observing arthritis score, surface temperature, color value and infrared imaging as well as ultrasonic blood flow signals in joints, as the evidences for syndrome model's judgment. Make a cross-validation with WuTou decoction and SiMiao powder respectively. Oxygen content in joints, expression of HIF, HSP-70 and citHSP in synovium, anti-collagen antibodies, anti-CCP antibody as well as a variety of levels of pro-inflammatory cytokines content was detected in serum, the ability of SF migration and erosion as well as peripheral lymphocytes responses are explored to find out the pathogenesis of model. This new combined model of disease and syndrome for RA can provide an important tool to study the mechanism of the TCM formulas with the advantages of differentiation of syndrome.

寒痹与热痹是RA的两个基本证型。本研究创新性地提出RASF增生是寒痹的主要病理，在此基础上出现炎症反应是热痹的主要病理。提出以RASF自发增生模型作为研究基础的新思路。TgTC鼠是本地培育的一种人肿瘤坏死因子转基因鼠（hTNF Tg），它能自发关节肿胀和畸形，不伴急性炎症反应，病理示SF高度增生而无炎症细胞浸润。拟以TgTC鼠作为基础模型，以Ⅱ型胶原免疫该鼠诱导免疫炎症反应，同时在人工气候箱里给予风寒湿或风湿热刺激。连续观察关节炎症积分，关节表面温度、色差值及红外成像，关节超声血流信号等作为证型判断依据；分别以乌头汤和四妙散进行交叉验证。检测关节含氧量，滑膜HIF、HSP-70以及citHSP的表达，血清抗胶原抗体、抗CCP抗体以及多种前炎症因子含量，SF迁移和侵蚀能力以及外周淋巴细胞的应答反应，以探讨模型的发病机制。这种新型病证结合模型能为体现RA中医辨证优势的方药机制研究提供新手段。

寒痹与热痹是RA的两个基本证型。本研究创新性地提出RASF增生是寒痹的主要病理，在此基础上出现炎症反应是热痹的主要病理。提出以RASF自发增生模型作为研究基础的新思路。TgTC鼠是本地培育的一种人肿瘤坏死因子转基因鼠（hTNF Tg），它能自发关节肿胀和畸形，不伴急性炎症反应，病理示SF高度增生而无炎症细胞浸润。拟以TgTC鼠作为基础模型，以Ⅱ型胶原免疫该鼠诱导免疫炎症反应，同时在人工气候箱里给予风寒湿或风湿热刺激。连续观察关节炎症积分，关节表面温度、色差值及红外成像，关节超声血流信号等作为证型判断依据；分别以乌头汤和四妙散进行交叉验证。检测关节含氧量，滑膜HIF、HSP-70以及citHSP的表达，血清抗胶原抗体、抗CCP抗体以及多种前炎症因子含量，SF迁移和侵蚀能力以及外周淋巴细胞的应答反应，以探讨模型的发病机制。关键结果数据：①人工气候箱风寒湿环境条件为：T：8℃，RH：90%，WS：5m/s；风湿热环境条件为：T：36℃，RH：90%，WS：5m/s。②Tgtc小鼠基础属性为寒，持续人工风寒湿环境干预时可作为寒痹模型，其发病实质与滑膜组织HIF1α上调有关；寒痹动物模型的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评估模型的客观量化指标。③风寒湿热环境刺激能影响CIA大鼠模型的中医证候；CIA大鼠基础属性为热，经FSR刺激后具有显著的热痹证候演变过程，可作为热痹模型，其实质与HSP70上调有关；经FHS刺激后并未表现寒痹证候特点，不能作为寒痹模型。④乌头汤通过上调滑膜组织及血清HSP70、下调滑膜组织HIF1α，延缓Tgtc小鼠关节炎进展，Tgtc小鼠属性偏寒，该模型病理实质为滑膜成纤维细胞增生为主，伴少量炎症细胞浸润，其机制可能与滑膜组织HIF1α上调有关。这种新型病证结合模型能为体现RA中医辨证优势的方药机制研究提供新手段。