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目的探讨p38丝裂原激活蛋白激酶(p38 MAPK)信号通路在脂多糖(LPS)诱导RAW264.7细胞表达可溶性髓样细胞触发受体1(s TREM-1)中的作用。方法培养小鼠巨噬细胞株RAW264.7,采用相同浓度的LPS在不同时间诱导RAW264.7细胞,应用Western blot法分别检测p38MAPK与磷酸化p38 MAPK(p-p38MAPK)蛋白表达水平,RT-PCR法检测p38 MAPK mRNA表达水平,酶联免疫吸附(ELISA)法检测细胞培养上清液中s TREM-1表达水平。用不同浓度p38 MAPK特异性抑制剂SB203580处理细胞,观察上述指标的变化。结果 LPS可呈时间依赖性诱导RAW264.7细胞p-p38 MAPK和p38 MAPK mRNA的表达;SB203580可呈浓度依赖性抑制p-p38 MAPK和p38 MAPK mRNA的表达;SB203580阻断p38 MAPK信号转导通路后,LPS对s TREM-1表达的诱导作用受到显著抑制,并且具有剂量依赖性。结论 LPS通过p38 MAPK信号通路诱导RAW264.7细胞表达s TREM-1。
Objective To investigate the role of the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in the induction of soluble myeloid cell-triggered receptor 1 (sTREM-1) in RAW264.7 cells induced by lipopolysaccharide (LPS). Methods Macrophage cell line RAW264.7 was cultured and RAW264.7 cells were induced at the same concentration of LPS at different times. Western blot was used to detect the expression of p38MAPK and p38 MAPK (p-p38MAPK) P38 MAPK mRNA was detected by enzyme-linked immunosorbent assay (ELISA), and s TREM-1 expression in cell culture supernatants was detected by ELISA. The cells were treated with SB203580, a specific inhibitor of p38 MAPK, and the changes of these indexes were observed. Results LPS induced the expression of p-p38 MAPK and p38 MAPK mRNA in RAW264.7 cells in a time-dependent manner. SB203580 inhibited the expression of p-p38 MAPK and p38 MAPK mRNA in a concentration-dependent manner. SB203580 blocked the p38 MAPK signal transduction pathway , The induction of sTREM-1 expression by LPS was significantly inhibited and in a dose-dependent manner. Conclusion LPS can induce sTREM-1 expression in RAW264.7 cells via p38 MAPK signaling pathway.