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目的探讨β2糖蛋白1(β2GP1)-抗β2GP1复合物诱导小鼠腹腔巨噬细胞表达肿瘤坏死因子α(TNF-α)中Toll样受体2(TLR2)的作用。方法用β2GP1-抗β2GP1复合物、TLR2激动剂Pam3CSK4及TLR4激动剂脂多糖(LPS)、TLR2阻断剂抗小鼠TLR2-Ig G(m TLR2-Ig G)及TLR4阻断剂TAK-242对BALB/c小鼠腹腔巨噬细胞进行体外处理。用实时荧光定量PCR检测TNF-αmRNA水平,Western blot法及免疫荧光细胞化学方法检测TNF-α蛋白表达;采用流式细胞术检测小鼠腹腔巨噬细胞表面TLR2的水平。结果β2GP1-抗β2GP1复合物、Pam3CSK4及LPS均能够显著增加BALB/c小鼠腹腔巨噬细胞表达TNF-αmRNA及蛋白水平,抗m TLR2-Ig G能够抑制上述刺激物促进细胞TNF-α表达的效应,但弱于TAK-242的抑制效应,抗m TLR2-Ig G与TAK-242联合使用未显示更强的抑制效应。流式细胞术结果显示,β2GP1-抗β2GP1复合物、Pam3CSK4及LPS均能增加小鼠腹腔巨噬细胞TLR2的表达,而抗m TLR2-Ig G、TAK-242以及二者联合使用均未显示对TLR2表达的抑制效应。结论TLR4、TLR2都增强β2GP1-抗β2GP1复合物对小鼠腹腔巨噬细胞TNF-α的刺激作用。
Objective To investigate the effect of β2GP1 - anti - β2GP1 complex on the expression of Toll - like receptor 2 (TLR2) in peritoneal macrophages induced by tumor necrosis factor - α (TNF - α) in mice. Methods The effects of anti-TLR2-Ig G (TLR2-Ig G) and TLR4 antagonist TAK-242 on TLR2 agonist Pam3CSK4 and TLR4 agonist lipopolysaccharide (LPS), TLR2 antagonist BALB / c mice peritoneal macrophages were treated in vitro. The level of TNF-αmRNA was detected by real-time fluorescence quantitative PCR. The expression of TNF-α protein was detected by Western blot and immunofluorescence cytochemistry. The level of TLR2 on the peritoneal macrophages was detected by flow cytometry. Results Both β2GP1-anti-β2GP1 complex, Pam3CSK4 and LPS could significantly increase the expression of TNF-αmRNA and protein in peritoneal macrophages of BALB / c mice. Anti-m TLR2-Ig G could inhibit the above stimuli to promote the expression of TNF-α Effect, but weaker than the inhibitory effect of TAK-242, the combination of anti-m TLR2-Ig G and TAK-242 did not show a stronger inhibitory effect. The results of flow cytometry showed that both β2GP1-anti-β2GP1 complex, Pam3CSK4 and LPS could increase the TLR2 expression in mouse peritoneal macrophages, while the combination of anti-TLR2-Ig G, TAK-242 and their combination did not show Inhibitory effect of TLR2 expression. Conclusion Both TLR4 and TLR2 enhance the stimulatory effect of β2GP1-anti-β2GP1 complex on TNF-α in mouse peritoneal macrophages.