类风湿关节炎寒证动物模型的建立与评价

Establishment and evaluation of cold syndrome animal model with rheumatoid arthritis

目的:复制符合中医证候特点的类风湿关节炎(RA)寒证动物模型并对其进行评价。方法:20只雌性Wistar大鼠,随机分为2组:一组复制胶原诱导性关节炎(CIA)大鼠模型;一组在复制CIA模型的基础上应用动物实验箱,从初次免疫开始连续28d给予风寒湿刺激。期间评价两组大鼠关节炎指数(AI)、足肿胀排水体积、关节疼痛阈值、外周关节体表温度、血流变血液黏度及凝血因子等。结果:两组发病率均为100%(10/10),CIA组成模率为75.00%(6/8),CIA寒证组成模率为77.78%(7/9),两组差异无统计学意义(χ2=0.0182,P=0.893)。但CIA寒证组左足排水体积在造模后14d与基线有差异,而CIA组在造模后21d与基线有差异(P<0.05)。两组大鼠造模后14d双足踝部肤温均升高,在造模后21d均有不同程度降低,组内比较两组各时间点差异均有统计学意义(P<0.05)。CIA寒证组在造模后21d左足体表温度显著低于CIA组(P<0.05)。两组后足关节的疼痛阈值在各个时间点差异无统计学意义;两组左足在造模后21d热板刺激反应时间缩短,疼痛阈值降低;在造模28d后疼痛阈值升高(P<0.05)。CIA寒证组中剪切率(50s-1、30s-1)和低剪切率(1s-1)下的血液黏度值显著升高(P<0.05)。CIA寒证组凝血酶凝结时间显著降低(P<0.05),其余剪切率下血液黏度和凝血因子在两组间差异无统计学意义。结论:CIA模型基础上联合风寒湿刺激,可以诱导出稳定可靠、符合中医证候特点的RA寒证动物模型。

Objective： To establish and evaluate a cold syndrome animal model with rheumatoid arthritis（RA） which conformed to the characteristics of TCM syndrome. Methods： 20 female Wistar rats were randomly divided into 2 groups： in one group, rats were copied into the collagen-induced arthritis（CIA） model; in the other group, rats were combined with the windcold-dampness stimulation starting from the initial immune for 28 d. The arthritis index（AI）, foot swelling drainage volume, joint pain threshold, peripheral joint surface temperature, blood viscosity of hemorhelogy and blood coagulation factor of two groups were evaluated. Results： The morbidity of two groups was 100%（10/10）, the success rate of molding in CIA group was 75.00%（6/8）, the success rate of molding in CIA with cold syndrome group was 77.78%（7/9）, there was no statistically significant differences between two groups（χ2=0.0182, P=0.0182）. While compared with the baseline, there was significant differences in the left foot drainage volume of CIA cold syndrome group in 14 d and the CIA group was in 21d（P〈0.05）. After modeling, the skin temperatures of ankle in two groups were increased in 14 d and decreased in 21 d, in each time points of two groups also had statistically significant differences（P〈0.05）. The left foot surface temperature of CIA cold syndrome group after modeling for 21 d was significantly lower than the CIA group（P〈0.05）. Foot joint pain threshold at all time points of two groups were no significantly differences. After modeling for 21 d, the hot-plate stimulus response time of left foot in two groups was shortened and the pain threshold was reduced; while the pain threshold increased after modeling for 28d（P〈0.05）. The blood viscosity values in middle shear rate（50/s, 30/s） and low shear rate（1/s） of CIA cold syndrome group were significantly increased（P〈0.05）. Thrombin time（TT） of CIA cold syndrome group was decreased significantly（P〈0.05）, there was no statistically significant difference between two groups on the blood viscosity in the rest shear rate and blood coagulation factors. Conclusion： The CIA model combined with the wind-cold-dampness stimulation could induce a stable and reliable cold syndrome animal model with RA which conformed to the characteristics of TCM syndrome. The TCM syndrome of CIA model was dynamic, it was possible to induce suitable animal model with TCM syndrome by applying different stimulations.