作者简介:丁利平(1984—),女,深圳大学博士后研究人员.研究方向:自身免疫及炎症. E-mail: 13552432187@163.com
中文责编:晨 兮; 英文责编:艾 琳
DOI: 10.3724/SP.J.1249.2016.06566
通过合成尿酸钠结晶(monosodium urate crystals, MSU)分析其对人外周血单核细胞系THP1的促炎作用,分别以 0、1.0、10.0和100.0 μmol/L的MSU刺激THP1细胞4 h,检测不同浓度的MSU对炎症体组分Nod 样受体蛋白3(Nod-like receptor protein 3, NLRP3)、半胱氨酸天冬氨酸特异性蛋白水解酶(caspase-1)及促炎因子白细胞介素-1β(interleukin-1β, IL-1β)和IL-18的信使核糖核酸(messenger ribonucleic acid, mRNA)表达的影响. 为研究toll 样受体4(toll like receptor 4,TLR4)的配体脂多糖的作用,用1.0 ng/mL的脂多糖(lipopolysaccharide, LPS)预刺激THP1细胞3 h,再加100.0 μmol/mL的MSU刺激细胞,通过实时荧光定量聚合酶链式反应及酶联免疫吸附试验,检测LPS对MSU诱导的促炎因子IL-1β和IL-18的mRNA及蛋白水平.研究发现,在THP1 细胞中,100.0 μmol/L的MSU即可显著激活炎症体的mRNA,但其对促炎因子IL-1β及IL-18的mRNA表达的诱导作用却不明显,加入1.0 ng/mL LPS预刺激3 h后,MSU诱导的THP1细胞IL-1β和IL-18的表达量显著增加.证明脂多糖对MSU诱导THP1细胞产生IL-1β 和IL-18有协同作用.
The synthesized monosodium urate crystals(MSU)was used for pro-inflammatory analyses in THP1 cells. THP1 cells were treated with 0, 1.0, 10.0 and 100.0 μmol/L MSU for 4 h in order to analyze the effects of MSU on the expression of messenger ribonucleic acid(mRNA)of inflammasome Nod-like receptor protein 3(NLRP3), caspase-1 and pro-inflammatory cytokines interleukin-1β(IL-1β)and IL-18. To investigate the role of lipopolysaccharide(LPS)-toll like receptor 4(TLR4)ligand, THP1 cells were pre-treated with 1.0 ng/mL LPS 3 h, and then stimulated with 100.0 μmol/mL MSU. The mRNA and protein expression of pro-inflammatory cytokines IL-1β and IL-18 induced by LPS and MSU were detected using quantitative real-time polymerase chain reaction(qPCR)and enzyme linked immunosorbent assay(ELISA), respectively. The results show that 100.0 μmol/L MSU could activate the mRNA expression of inflammasome, but has comparatively less effect on the mRNA expression of IL-1β and IL-18 in THP1 cells. The productions of IL-1β and IL-18 were significantly amplified in THP1 cells pretreated with 1.0 ng/mL LPS for 3 h before the MSU stimulation. Results suggest that MSU can act in synergy with LPS to enhance the release of IL-1β and IL-18 in mononuclear cells.