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目的:探讨Toll样受体4(TLR4)和巨噬细胞移动抑制因子(MIF)对ECV304内皮细胞增殖、黏附及炎症因子水平的影响。方法:ECV304细胞经TLR4 MIF单克隆抗体(mAb)和脂多糖(LPS)处理后,通过3H-TdR掺入实验、黏附实验和放射免疫分析研究细胞增殖、黏附和细胞因子水平。结果:LPS(10 mg/L)明显促进ECV304细胞TLR4的表达,而抗MIF mAb(10、50 mg/L)明显抑制其表达。抗MIF(50 mg/L)和抗TLR4mAb(1 mg/L)明显抑制了ECV304细胞的增殖,而LPS(10 mg/L)明显促进增殖。抗MIF mAb明显抑制了人外周血淋巴细胞对ECV304细胞的黏附,而LPS和抗TLR4 mAb对黏附无显著影响。抗MIF、抗TLR4 mAb明显抑制ECV304细胞TNFα的分泌,并呈剂量依赖,而LPS作用相反。与对照组相比,LPS(1 mg/L,48 h)可以促进ECV304细胞IL-8的分泌,抗MIF mAb(50 mg/L,48 h)可以明显抑制IL-8的分泌(P<0.05)。但是抗TLR4 mAb(1 mg/L)对ECV304细胞IL-8的分泌无明显影响。结论:LPS、抗TLR4及抗MIF mAb对血管内皮细胞TLR4表达及活性有显著影响。
Objective: To investigate the effects of Toll-like receptor 4 (TLR4) and macrophage migration inhibitory factor (MIF) on the proliferation, adhesion and inflammatory cytokines in ECV304 endothelial cells. Methods: ECV304 cells were treated with TLR4 MIF monoclonal antibody (mAb) and lipopolysaccharide (LPS), and cell proliferation, adhesion and cytokine levels were studied by 3H-TdR incorporation assay, adhesion assay and radioimmunoassay. Results: LPS (10 mg / L) significantly promoted TLR4 expression in ECV304 cells, while anti-MIF mAb (10,50 mg / L) significantly inhibited the expression of TLR4. Anti-MIF (50 mg / L) and anti-TLR4 mAb (1 mg / L) significantly inhibited ECV304 cell proliferation, whereas LPS (10 mg / L) significantly promoted proliferation. Anti-MIF mAbs significantly inhibited the adhesion of human peripheral blood lymphocytes to ECV304 cells, whereas LPS and anti-TLR4 mAbs had no significant effect on adhesion. Anti-MIF, anti-TLR4 mAb significantly inhibited the secretion of TNFα in ECV304 cells in a dose-dependent manner, while LPS had the opposite effect. Compared with the control group, LPS (1 mg / L, 48 h) could promote the secretion of IL-8 in ECV304 cells. Anti-MIF mAb (50 mg / L, 48 h) ). However, anti-TLR4 mAb (1 mg / L) had no significant effect on IL-8 secretion in ECV304 cells. Conclusion: LPS, anti-TLR4 and anti-MIF mAb have significant effects on TLR4 expression and activity in vascular endothelial cells.