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目的探讨可溶型树突状细胞特异性的结合细胞间粘附分子的非整合素(sDC-SIGN)对脂多糖(LPS)刺激的单核细胞炎症反应的调节作用。方法以人急性单核细胞白血病细胞株(THP-1)为研究对象,分为空白组、200 ng/ml sDC-SIGN单独处理组、LPS单独刺激组、LPS刺激同时加100 ng/ml sDC-SIGN处理组和LPS刺激同时加200 ng/ml sDC-SIGN处理组,刺激后不同时间点收集上清和细胞。ELISA检测细胞上清中细胞因子含量、Q-PCR检测细胞内细胞因子的mRNA表达并利用Western blot方法检测细胞内细胞外调节蛋白激酶(ERK)和核转录因子-κB(NF-κB)信号通路的活化状况。结果不同浓度的sDC-SIGN均可抑制LPS刺激炎性细胞因子TNF-α、IL-6和IFN-α的mRNA表达和蛋白分泌,同时促进抑炎细胞因子IL-10的mRNA表达和蛋白分泌,且sDC-SIGN的抑制效果与浓度呈依赖关系;研究结果亦显示,sDC-SIGN减弱LPS刺激THP-1细胞诱导ERK和NF-κBP65的磷酸化水平。结论sDC-SIGN蛋白对LPS诱导的单核细胞炎症应答具有负向调控作用。
Objective To investigate the regulatory effect of soluble dendritic cell-specific non-integrin (sDC-SIGN), which binds intercellular adhesion molecule, on the inflammatory response of monocytes stimulated by lipopolysaccharide (LPS). Methods Human acute monocytic leukemia cell line (THP-1) was divided into blank group, 200 ng / ml sDC-SIGN single treatment group, LPS stimulation group and LPS stimulation plus 100 ng / ml sDC- SIGN treatment group and LPS stimulation plus 200 ng / ml sDC-SIGN treatment group, stimulated at different time points after the supernatant and cells were collected. The levels of cytokines in the supernatant of the cells were detected by ELISA. The mRNA expression of cytokines was detected by Q-PCR. The expression of extracellular regulated protein kinase (ERK) and nuclear factor kappa B (NF-κB) Activation status. Results Different concentrations of sDC-SIGN could inhibit the mRNA and protein secretion of inflammatory cytokines TNF-α, IL-6 and IFN-α induced by LPS, at the same time, promote the mRNA and protein secretion of pro-inflammatory cytokines IL-10, And the inhibitory effect of sDC-SIGN was dependent on the concentration. The results also showed that sDC-SIGN attenuated the phosphorylation of ERK and NF-κB P65 induced by LPS in THP-1 cells. Conclusion The sDC-SIGN protein negatively regulates the LPS-induced monocyte inflammatory response.