类风湿性关节炎大鼠模型的构建及免疫相关基因mRNA表达的研究

Establishment of rat model of rheumatoid arthritis and the mRNA expression of immune related genes

为了比较类风湿性关节炎大鼠模型建模效果,并探究免疫相关基因mRNA的表达,本研究分别采用胶原(CIA)和完全弗氏佐剂(AA)两次皮内注射方法制备大鼠模型,对照组(C组)注射等体积生理盐水,以足趾肿胀值判断成模情况。在造模后不同时间点,采用ELISA试剂盒测定血清中细胞因子IL-1β的含量,运用RT-PCR方法检测脾组织中免疫相关基因PD-1、TGF-β1和Foxp3 mRNA的表达。结果显示,AA组和CIA组大鼠足趾肿胀值均显著增大(CIA>AA>对照组),血清IL-1β浓度在第2周与第4周显著升高(CIA>AA>对照组),脾组织中PD-1、TGF-β1和Foxp3 mRNA表达显著降低(CIA<AA<对照组)。以上结果表明,CIA和AA注射均可成功制备RA模型,且CIA模型较AA模型成模效果更好,IL-1β在RA发病中起着至关重要的作用。

In order to compare modeling effect of rats with rheumatoid arthritis and to explore the mRNA expression of immune related genes.In the present study,intradermal injection of collagen(CIA)and complete freund's adjuvant(AA)were used to establish the RA rat model,while the control rats were injected with equal volume of physiological saline.Formation of toe swelling value was used to judge the success of the model.Furthermore,ELISA was used to detect the IL-1βlevel in serum and the mRNA expression of immune related genes PD-1,TGF-β1 and Foxp3 in spleen tissue were determined by RT-PCR at different time points after operation.Compared with control group,formation of toe swelling value in CIA and AA group were increased significantly,in addition,serum IL-1βconcentrations of CIA and AA rats in the second and fourth weeks of testing were higher significantly.PD-1,TGF-β1 and Foxp3 mRNA expression in the spleen were significantly lower in CIA and AA rats after four weeks of modeling.And all above changes in CIA rats were more obvious than AA rats.The above results showed that both methods can successfully establish rat model of rheumatoid arthritis,CIA method is better than AA,and IL-1βplays an important role in the progress of RA.