两种佐剂制备CIA大鼠模型的效果比较

郭帅; 褚小磊; 沈桐; 李季; 王浩淼; 朱毛妮; 朱盼盼; 陈一航; 吕樵岚; 胡玲莉; 田亚兰; 王元元; 陈传好

doi:10.13898/j.cnki.issn.1000-2200.2022.03.001

摘要:

目的比较不完全弗氏佐剂(incomplete Freund′s adjuvant，IFA)和完全弗氏佐剂(complete Freund′s adjuvant, CFA)分别与牛Ⅱ型胶原蛋白(calf Ⅱ collagen，CⅡ)构建的胶原诱导的类风湿性关节炎(collagen-induced rheumatoid arthritis，CIA)大鼠模型的效果。

方法选取30只8周龄SD大鼠，随机分为0.9%氯化钠溶液组(对照组)、IFA+CⅡ组和CFA+CⅡ组，每组10只。将IFA/CFA与CⅡ以等体积混匀，分别从鼠尾根部进行皮内注射，在第0天和第7天每只鼠分别注射200 μL和100 μL。采用相同方法注射0.9%氯化钠溶液作为对照组。从初次免疫开始，每7天记录1次关节炎指数(arthritis index，AI)分值。在第14天分别从3组中取5只大鼠获取踝关节，做HE染色并观察病理学变化。

结果造模第0天和第7天，3组AI值为0；初次免疫后第14天，IFA+CⅡ组和CFA+CⅡ组的AI均值分别为(6.5±0.5)和(7.4±0.4), 2组差异有统计学意义(P < 0.01)；造模后第21天，IFA+CⅡ组和CFA+CⅡ组的AI均值分别为(8.8±0.5)和(7.8±0.8)，2组差异有统计学意义(P < 0.01)；造模后第14天和第21天，2组差异比较均无统计学意义(P>0.05)。踝关节HE染色结果发现，IFA+CⅡ组踝关节软骨表面粗糙不平，轻微破坏；关节腔内可见炎性细胞浸润。CFA+CⅡ组关节软骨破坏明显, 呈锯齿状，有明显的血管翳和大量的炎性细胞浸润。

结论使用IFA+CⅡ和CFA+CⅡ均可制备RA模型，IFA+CⅡ的炎症反应较轻，为制备RA模型的首选方案。

Abstract:

ObjectiveTo compare the effects of incomplete Freund′s adjuvant(IFA) and complete Freund′s adjuvant(CFA) combined with calf Ⅱ collagen(CⅡ) in establishing the collagen-induced rheumatoid arthritis(CIA) rat model.

MethodsThirty SD rats aged 8 weeks were randomly divided into the 0.9% sodium chloride solution group(control group), IFA+CⅡ group and CFA+CⅡ group(10 rats in each group).The IFA/CFA and CⅡ were blerding in equal volume, and intradermally injected into the root of rat tail.The 200 μL and 100 μL mixture were injected into rats in the model group on day 0 and day 7, respectively.The same volume of 0.9% sodium chloride solution was injected into rats in the control group using the same method.The scores of arthritis index(AI) in three groups were recorded every 7 days from the initial immunization.HE staining was performed in ankle joints of 5 rats from three groups on day 14 to observe the pathological changes.

ResultsOn day 0 and day 7 after modeling, the AI values in three groups were 0.On day 14 after initial immunization, the AI values in IFA+CⅡ group and CFA+CⅡ group were(6.5±0.5) and(7.4±0.4), respectively, and the difference of which was statistically significant(P < 0.01).On day 21 after modeling, the AI values in IFA+CⅡ group and CFA+CⅡ group were(8.8±0.5) and(7.8±0.8), respectively, and the difference of which was statistically significant(P < 0.01).But the difference of AI values in IFA+CⅡ group and CFA+CⅡ group were not statistically significant on day 14 and day 21 after modeling(P>0.05).The results of HE staining of the ankle joint showed that the cartilage surface in ankle joint in the IFA+CⅡ group was rough and slightly damaged, and inflammatory cell infiltration was observed in the articular cavity.In CFA+CⅡ group, the articular cartilage was obviously damaged, presented saw tooth shape, and had obvious pannus and a large amount of inflammatory cell infiltration.

ConclusionsBoth IFA+CⅡ and CFA+CⅡ can be used to prepare RA model, and the inflammatory response of IFA+CⅡ is slight, which is the preferred method for preparing RA model.

两种佐剂制备CIA大鼠模型的效果比较

Effect comparison of two adjuvants in the preparation of rat CIA model