类风湿关节炎血瘀证大鼠模型的制备及评价

目的 :建立一个稳定可靠的类风湿关节炎(RA)血瘀证动物模型,为后续实验提供基础。方法:选用雌性SD大鼠,在Ⅱ型胶原诱导的关节炎模型(CIA)的基础上,结合小剂量长期注射盐酸肾上腺素和4℃冷水应激的方法,建立RA血瘀证动物模型并对其进行评价。结果:同正常组对比,模型组一般精神状况较差,足趾肿胀度在各个时间点比较差异有统计学意义(P<0.01)。模型组大鼠造模后第7天至第28天关节炎指数(AI)值持续增高,在各个时间点与正常组对比差异有统计学意义(P<0.01)。模型组大鼠外周血血清类风湿因子(RF)、肿瘤坏死因子(TNF-α)、血栓素A2(TX-A2)、超敏C反应蛋白(hs CRP)的浓度水平较正常组升高(P<0.01),且4周内呈持续升高趋势。模型组大鼠高中低切变率下的全血黏度及血浆黏度均高于正常组(P<0.05或P<0.01);模型组红细胞压积高于正常组(P<0.01);模型组血沉快于正常组(P<0.01)。光镜下可见模型组大鼠关节软骨表面不光整,浅表层可见增生纤维化,部分可见裂纹。软骨全层细胞减少,细胞排列紊乱,分布不均,部分细胞核坏死,部分软骨陷窝消失。软骨基质染色浅且不均匀。潮线模糊、不规则,附近有血管穿过。模型组病理评分分值较正常组升高(P<0.01)。结论:本研究在CIA模型的基础上,结合盐酸肾上腺素小剂量长期注射复合4℃冷水应激,制备的模型更符合RA血瘀证久病、慢性的特点,更能接近临床血瘀证的发病过程,符合中医证候特点。

Preparation and Evaluation about Rat Models with Blood Stasis Syndrome of Rheumatoid Arthritis

Objective:The study intends to establish a stable and reliable animal model with blood stasis syndrome of rheumatoid arthritis(RA) and provide the basis for further experiments.Methods:The female SD rats were selected and on the basis of the CIA arthritis model,by long-term injection with small dose of adrenaline hydrochloride and 4 ℃ cold water stress,to establish animal models with blood stasis syndrome of RA.Results:Compared with the normal group,the model group were generally poor mental health,toe swelling degrees at all time points had significant differences (P< 0.01).From 7 d to 28 d after the model group rats are built,its arthritis index(AI) value continued to increase and had differences compared to the normal group (P< 0.01) at all time points.Levels of RF,TNF-α,TX-A2 and hs CRP in the model group were increased compared to the normal group (P< 0.01) and showed a trend of rising within 4 weeks.Shear rate of whole blood viscosity and plasma viscosity in the model group rats were higher than that in the normal group (P< 0.05 or 0.01).Hematocrit of the model rats was higher than that of the normal group (P< 0.01).Blood sedimentation of the model group was faster than that of the normal group (P< 0.01).The unsmoothness,visible hyperplasia,fibrosis and part of cracks could be seen on the articular cartilage surface of the model group under microscope.Full-thickness cartilage cells decreased,irregular arrangement of cells,uneven distribution,part of the nucleus necrosis,part of the cartilage pit disappeared.Cartilage matrix was dyed lightly and unevenly.Fuzzy and irregular tide line,there were blood vessels crossing through.The pathological grading score of the model group was higher than that in the normal group (P< 0.01).Conclusion:The model rats based on the CIA model,combined with small dose of adrenaline hydrochloride injection and 4 ℃cold water stress for a long time,were more suitable to "long illness" and "chronic" characteristics of blood stasis syndrome of RA,more close to the clinical blood stasis process and TCM syndrome characteristics.