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目的探讨Toll样受体2(TLR2)配体肽聚糖(PGN)和脂磷壁酸(LTA)对THP-1细胞表达趋化因子的影响。方法 THP-1细胞用不同浓度(1、10、100μg/m L)PGN或LTA刺激后,收集细胞提取m RNA进行实时荧光定量PCR检测,收集培养上清进行ELISA检测。抑制实验中,THP-1细胞先用信号分子抑制剂预处理,与PGN或LTA孵育后,再进行实时荧光定量PCR检测。结果刺激6 h,浓度为10μg/m L的PGN和LTA能显著促进CCL2、CXCL8和CXCL10 m RNA的表达(P<0.05),当浓度提高至100μg/m L时,PGN和LTA能更显著促进CCL2、CXCL8和CXCL10 m RNA的表达(P<0.001);刺激24 h,浓度为10μg/m L的PGN或LTA能显著促进THP-1细胞释放趋化因子CCL2、CXCL8和CXCL10;不同信号分子抑制剂有差异地抑制PGN或LTA介导上调的CCL2、CXCL8和CXCL10的表达。结论 TLR2配体PGN和LTA促进THP-1细胞表达趋化因子CCL2、CXCL8和CXCL10 m RNA,受信号通路MPAK和NF-κB差异调控。
Objective To investigate the effect of Toll-like receptor 2 (TLR2) ligand peptidoglycan (PGN) and lipoteichoic acid (LTA) on the expression of chemokines in THP-1 cells. Methods After THP-1 cells were stimulated with PGN or LTA at different concentration (1, 10, 100μg / mL), m RNA was collected and detected by real-time fluorescence quantitative PCR. The supernatant was collected for ELISA. Inhibition experiments, THP-1 cells pretreated with a signal inhibitor, with PGN or LTA incubation, and then real-time fluorescence quantitative PCR detection. Results PGN and LTA at a concentration of 10 μg / mL for 6 h significantly promoted the expression of CCL2, CXCL8 and CXCL10 m RNA (P <0.05). When the concentration was increased to 100 μg / mL, PGN and LTA could significantly promote CCL2, CXCL8 and CXCL10 m RNA expression (P <0.001); PGN or LTA at a concentration of 10 μg / mL for 24 h could significantly promote the release of chemokines CCL2, CXCL8 and CXCL10 by THP-1 cells; Agents differentially inhibited PGN or LTA-mediated up-regulation of CCL2, CXCL8 and CXCL10 expression. Conclusion The TLR2 ligands PGN and LTA can promote the expression of chemokines CCL2, CXCL8 and CXCL10 m RNA in THP-1 cells, which are regulated by the difference between MPAK and NF-κB signaling pathways.