摘要
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 role of Toll-like receptor 2（ TLR2） in β2-glycoprotein 1/anti-β2-glycoprotein 1（ β2GP1 /anti-β2GP1）-mediated tumor necrosis factor α（ TNF-α） expression in mouse peritoneal macrophages. Methods The peritoneal macrophages from BALB / c mice were treated with β2GP1 / anti-β2GP1 complex, agonist of TLR2（ Pam3CSK4） and agonist of TLR4 [lipopolysaccharide（ LPS） ],inhibitor of TLR2 [monoclonal Ig G to mouse TLR2（ anti-m TLR2-Ig G） ]and inhibitor of TLR4（ TAK-242） in vitro. The mRNA level of TNF-α in the peritoneal macrophages was tested by real-time quantitative PCR, the protein expression of TNF-α was detected by Western blotting and immunofluorescence cytochemistry,and the expression of TLR2 on the surface of the peritoneal macrophages was assessed by flow cytometry. Results The mRNA and protein expression of TNF-α was significantly enhanced in mouse peritoneal macrophages treated with β2GP1 / anti-β2GP1 complex,Pam3CSK4 and LPS. Anti-m TLR2-Ig G could inhibit the effects of the above stimuli on TNF-α expression,but its effects were weaker than those of TAK-242. Meanwhile,the combination of anti-m TLR2-Ig G and TAK-242 did not show much stronger inhibitory effects. Flow cytometry analysis showed that expression of TLR2 was enhanced by β2GP1 / anti-β2GP1 complex,Pam3CSK4 and LPS in mouse peritoneal macrophages. However,anti-m TLR2-Ig G,or TAK-242,or combination of both could not inhibit TLR2 expression in macrophages. Conclusion Both TLR4 and TLR2 could increase the stimulating effect of β2GP1 / anti-β2GP1 complex on the expression of TNF-α in mouse peritoneal macrophages.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第4期446-450,456,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81370614)
