类风湿关节炎小鼠模型中Bregs细胞、IL-10和TGF-β的变化及其内在关系

Changes of Bregs cells,IL-10 and TGF-β in mouse model of rheumatoid arthritis and their internal relations

目的:探讨Bregs细胞、血清中IL-10和TGF-β与类风湿关节炎(RA)活动的相关性。方法:建立小鼠Ⅱ型胶原诱导类风湿关节炎(CIA)模型;采用HE染色观察对照组和CIA组小鼠关节滑膜和周围细胞的变化,并提取滑膜组织中的mRNA检测其中IL-10和TGF-β的变化。采用CBA法测定CIA组与对照组血清中IL-10、TGF-β的含量。采用流式细胞术检测CIA组与对照组中Breg细胞(CD1dhighCD5+CD19high)比例的变化。结果:对照组小鼠足爪关节无红肿;CIA组小鼠足跖部出现肿胀,关节明显红肿增粗。CIA组与对照组关节炎评分差异显著(P<0.05)。对照组小鼠滑膜细胞排列整齐,无增生,无炎症细胞浸润;CIA组滑膜细胞水肿,排列不整齐,有3~5层明显增生,有大量的炎症细胞浸润,并出现新生血管。与对照组相比,CIA组滑膜细胞中IL-10和TGF-β的mRNA表达水平差异显著(P<0.05)。同时,CIA组血清中IL-10和TGF-β的含量较对照组显著升高(P<0.05)。与对照组相比,CIA组小鼠脾脏中Breg细胞(CD1dhighCD5+CD19high)比例明显增多(P<0.05)。结论:与对照组相比,在CIA组小鼠血清和滑膜组织中IL-10和TGF-β表达水平、脾脏中Breg细胞比例均显著升高,Breg细胞在RA发病进程中起重要作用。

Objective:To investigate the correlation Bregs cells and IL-10 and TGF-β in serum in rheumatoid arthritis(RA) animal model.Methods: Collagen-induced arthritis(CIA) mouse model was established.The synovial membrane and surrounding cells were observed by HE staining both in the mice of control group and CIA group.The variation of IL-10 and TGF-β at mRNA level in the synovial membrane were detected.The serum levels of IL-10 and TGF-β in the CIA group and the control group were also determined by CBA method.The change of the ratio of regulatory B cells(Bregs) cells(CD1 dhighCD5+CD19high) in the CIA group and the control group was detected by flow cytometry.Results: There was no redness and swelling in the paw joints of the mice in the control group.The swelling of the foot and ankle in the CIA group showed obvious redness and thickening of the joints.There was significant difference in arthritis score between CIA group and control group(P<0.05).In the control group,the synovial cells were arranged neatly,without hyperplasia and no inflammatory cell infiltration;the synovial cells in the CIA group were edema,irregularly arranged;3 to 5 layers were obviously proliferated,a large number of inflammatory cells infiltrated,and new blood vessels appeared.Compared with the control group,the mRNA levels of IL-10 and TGF-β in the synovial cells of the CIA group were significantly different(P<0.05).At the same time,the levels of IL-10 and TGF-β in serum of the CIA group were significantly higher than those in control group(P<0.05).Compared with the control group,the proportion of Breg cells(CD1 dhighCD5+CD19high) in the spleen of the CIA group was significantly increased(P<0.05).Conclusion: Compared with the control group,the expression levels of IL-10 and TGF-β in serum and synovial tissue and the proportion of Breg cells in spleen in CIA mouse model were significantly increased,Breg cells play an important role in the pathogenesis of rheumatoid arthritis.