目的 探讨肿瘤坏死因子α诱导蛋白3(TNFAIP3)对MRL/lpr小鼠B淋巴细胞功能的影响，进一步探索其在免疫及炎症反应中发挥功能的具体作用机制，为系统性红斑狼疮(SLE)的临床研究提供实验依据。方法 采用腺相关病毒(AAV)感染提高MRL/lpr小鼠TNFAIP3的表达，RT-qPCR及 Western blot检测B淋巴细胞中TNFAIP3、Toll样受体7(TLR7)、Toll样受体9(TLR9)基因及蛋白表达水平，ELISA法检测血清抗ds-DNA抗体、TNF-α和IL-6的水平，分析TNFAIP3与TLR7/TLR9之间的关系。结果 成功过表达MRL/lpr小鼠B淋巴细胞的TNFAIP3后，该组小鼠血清中抗ds-DNA抗体、TNF-α和IL-6的水平与对照组相比明显降低(P＜0。05)。Western blot及RT-qPCR检测显示TLR7、TLR9蛋白及mRNA水平与对照组相比明显降低(P＜0。05)。结论 TNFAIP3可能通过抑制B淋巴细胞中TLR7/TLR9蛋白及mRNA表达水平，抑制MRL/lpr小鼠体内自身抗体和炎症因子的产生，从而缓解MRL/lpr小鼠的病情进展。
Effects of TNFAIP3 expression level on B lymphocytes function and inflammatory response in MRL/lpr mice
Objective To investigate the effect of tumor necrosis factor α-induced protein 3(TNFAIP3)on the function of B lymphocytes in MRL/lpr mice,and further explore its specific mechanisms of action in immune and inflammatory responses,providing experimental basis for clinical research on systemic lupus erythematosus(SLE).Methods Adeno-associated virus(AAV)was used to increase the expression of TNFAIP3 in MRL/lpr mice.RT-qPCR and Western blot were used to detect the expression levels of TNFAIP3,Toll-like receptor 7(TLR7)and Toll-like receptor 9(TLR9)in B lymphocytes.The levels of serum anti ds DNA antibodies,TNF-α and IL-6 were measured by ELISA.Results After successful overexpression of TNFAIP3 in the experimental group,the serum levels of anti ds DNA antibodies,TNF-α,and IL-6 in the experimental group were significantly decreased compared with the control group(P＜0.05).Westernblot and RT-qPCR showed that the protein and mRNA expression levels of TLR7 and TLR9 in the experimental group were significantly decreased compared with the control group(P＜0.05).Conclusion TNFAIP3 may alleviate the progression of MRL/lpr mice by inhibiting the expression levels of TLR7/TLR9 protein and mRNA in B lymphocytes,inhibiting the production of autoantibodies and inflammatory factors in MRL/lpr mice.
万方数据
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