摘要
从类风湿性关节炎(RA)患者血清中分离抗环瓜氨酸化抗体(ACPAs),观察ACPAs对巨噬细胞功能的影响,探讨ACPAs影响RA病情发展的可能机制。利用亲和层析从RA患者血清中分离ACPAs,通过荧光定量PCR、蛋白印迹(Western blotting)及流式细胞术等实验方法观察ACPAs对巨噬细胞功能的影响。结果表明:亲和层析可很好地从血清中分离ACPA。ACPAs(40 000 IU/L)可促进巨噬细胞肿瘤坏死因子α(TNFα)、白细胞介素6(IL-6)、诱导型一氧化氮合成酶(iNOS)、人类白细胞DR抗原(HLA-DR)、磷酸化蛋白激酶B(p-AKT)等炎性因子的表达。SiRNA转染实验证实:ACPAs通过促进AKT蛋白磷酸化,诱导TNFα、IL-6、iNOS的表达。
Anti-citrullinated protein antibodies(ACPAs)were isolated from rheumatoid arthritis(RA)patient,and the mechanism of ACPAs-induced macrophage activities was studied.ACPAs were purified by affinity column.Fluorescence-activated cell sorting,Western blotting,and real-time PCR analyses were performed to detect the ACPAs-induced macrophage action.Results show that ACPAs(40 000 IU/L)could induce tumor necrosis factor α(TNFα),interleukin(IL)-6,inducible nitric oxide synthase(iNOS),human leukocyte antigen(HLA)-DR and phosphorylated protein kinase B(p-AKT)expression in macrophage.ACPAs increased TNFα,IL-6 and iNOS expression through promoting p-AKT expression.
引文
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