改良痛风性关节炎大鼠模型的复制

Replication of Rat Model of Gouty Arthritis

目的复制腹腔连续注射氧嗪酸钾结合Coderre法的改良痛风性关节炎大鼠模型的方法。方法 40 SD大鼠分为4组,每组10只,A组连续腹腔注射生理盐水1周后,在右侧踝关节腔注射生理盐水;B、C组连续腹腔注射氧嗪酸钾1周后,B组在右侧踝关节腔注射尿酸钠溶液,C组注射生理盐水;D组连续腹腔注射生理盐水1周后,在大鼠右侧踝关节腔注射尿酸钠溶液。造模期间观察大鼠的饮食、精神状态、大小便等情况,于造模前、造模后4、12、24、48h测量关节肿胀程度,检测造模48h后血尿酸、观察踝关节滑膜组织病理变化及检测滑膜组织中TNF-α、IL-1β、IL-6含量。结果 (1)造模前,各组大鼠皮毛色白有光泽,精神佳,活泼好动,进食及饮水正常,体重增长正常;造模后,A组饮食、饮水、活动及大便均正常,踝关节无明显肿胀及活动减少;B组大鼠出现不同程度毛色干枯发黄、脱落,进食、饮水减少,踝关节红胀、肤温增高,活动明显减少;C组大鼠出现不同程度毛色干枯发黄、脱落,进食、饮水减少,踝关节无明显肿胀及活动减少;D组饮食、饮水、活动及大便均基本正常,踝关节红胀、肤温增高,活动明显减少。(2)关节肿胀指数变化结果:与A组相比,B、D组各时点肿胀指数明显升高(P<0.01),C组差异无统计学意义(P>0.05)。(3)48h后血尿酸变化结果与A组比较,B、C组大鼠血尿酸值均明显升高(P<0.01,P<0.01),D组差异无统计学意义(P>0.05)。(4)踝关节滑膜组织病理改变:A、C组解剖时踝关节及其周围组织结构正常清晰,无任何组织病理学改变,光镜见滑膜上皮排列完整,滑膜下血管分布正常,无血管增生,无炎症细胞浸润,B、D组局部解剖时有尿酸盐结晶沉着于踝关节腔内,可见明显的关节炎病理改变,滑膜上皮脱落,甚至消失不见,见大量炎症细胞(主要是中性粒细胞),伴有血管增生(见图4B)。(5)滑膜组织中炎性因子(TNF-α、IL-1β、IL-6)水平比较:与A组比较,B、D组大鼠滑膜组织中TNF-α、IL-1β、IL-6水平均显著增高,差异有统计学意义(P<0.01,P<0.01)(见表4),C组无明显差异(P>0.01)。结论腹腔连续注射氧嗪酸钾结合Coderre法的改良急性痛风性关节炎大鼠模型可以成功复制,该模型较符合于人类自然病程,即痛风性关节炎建立在血尿酸升高生化基础上的,可有效排除尿酸分解酶对关节腔中尿酸钠晶体的影响,且经济、可重复性较高,可作为痛风性关节炎研究的复合型动物模型。

Objective To establish a method for the continuous improvement of rat model of gouty arthritis by intraperitoneal injection of potassium oxygen and Coderre. Methods 40 healthy adult male SD rats were randomly divided into four groups, each group of 10. Group B, group C continuous intraperitoneal injection of oxygen oxazine acid potassium after 1 week, group B on the right side of the ankle joint cavity injection in rats, uric acid sodium solution group C equal dose of normal saline injection;Group A and group D continuous intraperitoneal injection of saline solution after 1 week, group D on the right side of the ankle joint cavity injection in rats, uric acid sodium solution group A equal dose of normal saline injections. Observed in rats during the period of building diet, mental state, urine, etc. Prior to the building, the building after 4 h, 12 h, 24 h, 48 h measuring joint swelling degree, detection of building after 48 h blood uric acid, observe ankle synovial tissue pathological changes and test the synovial tissue of TNF-α, IL-1β, IL-6. Results （1） Building, before each rat fur color white luster, good spirit, active and lively, to eat and drink is normal, normal weight gain; After building, food, drinking water, activities and group A shit all normal, no obvious swelling in the ankle and decreased activity ; B group of rats appeared different degree of colour dry yellow, fall off, eat, drink less, ankle red swelling, skin temperature, activity decreased significantly; Group C rats appeared different degree of colour dry yellow, fall off, eat, drink less, no obvious swelling in the ankle and decreased activity; Food, drinking water, activities and defecate D group were essentially normal, ankle red swelling, skin temperature, activity decreased significan@. （2） Joint swelling index change results： compared with group A, group B and D each point swell index increased significandy （P〈0.01）, there was no statistically significant difference in group C （P〉0.05）. （3） The blood uric acid changes after 48 results compared with group A, B and C group rats blood uric acid values were significantly iucreased（P〈0.01, P〈0.01 ）, there was no statistically significant difference group D （P〉0.05）. （4） The ankle joint synovial tissue pathological changes： A group C, the normal anatomy of the ankle and around the organization structure is clear, no histopathological changes, arrange complete lens see the synovial epithelium, synovial vascular distribution under normal, no blood vessel growth, no inflammatory cell infiltration, group B and D local anatomic urate crystals composed in ankle joint cavity, clearly visible pathological changes of arthritis, synovial epithelial shedding, even disappear, see A large number of inflammatory cells, mainly neutrophils ）, accompanied by blood vessel growth （see figure 4 B）. （5） In the synovial tissue inflammatory factors（TNF alpha, beta, IL-6, IL-1） horizontal comparison： compared with group A, group B and D in the synovial tissue of rats TNF alpha, beta, IL-6, IL -1 levels were significantly increased, the difference was statistically significant （P〈 0.01, P〈 0.01 ） （ see table 4 ）, no obvious difference was found between group C （P〉 0.01 ）. Conclusion The combination of abdominal continuous injection of oxygen oxazine acid potassium Coderre method improved gouty arthritis rat model can be successfully copied. The model is more consistent with the natural course of human beings, and can effectively eliminate the influence of the uric acid decomposing enzyme on the sodium urate crystal in the joint cavity. The model can be used as a composite animal model for the study of gouty arthritis.