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目的研究咖啡酸(caffeic acid,CA)对同型半胱氨酸(homocysteine,Hcy)诱导的小鼠外周血白细胞炎症介质分泌的影响。方法分离8周龄雄性C576J/BL小鼠外周血白细胞,用Hcy(200μmol/L)和CA(100μmol/L)处理30 min。用流式微球法检测白细胞培养基中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)的水平。用激光共聚焦显微镜观察白细胞中髓过氧化物酶(myeloperoxidase,MPO)的表达。用western blot检测白细胞中磷酸化的细胞外信号调节激酶(phospho-extracellu-lar signal-regulated protein kinase 1/2,p-ERK1/2)蛋白的表达。结果小鼠外周血白细胞用Hcy处理后,培养基中TNF-α水平没有显著的变化,而MCP-1水平显著升高,白细胞MPO分泌增加,p-ERK1/2蛋白表达显著增加。CA抑制了Hcy诱导的小鼠外周血白细胞炎症介质MCP-1和MPO的分泌,以及p-ERK1/2蛋白表达的增加。结论CA可以抑制Hcy诱导的小鼠外周血白细胞MCP-1和MPO的释放,该作用可能与其抑制p-ERK1/2的表达有关。
Objective To investigate the effect of caffeic acid (CA) on the secretion of interleukin (IL) in mice induced by homocysteine (Hcy). Methods Peripheral blood leucocytes from male C576J / BL mice of 8 weeks old were isolated and treated with Hcy (200μmol / L) and CA (100μmol / L) for 30 min. Flow cytometry was used to detect the levels of tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in leukocyte culture medium. The expression of myeloperoxidase (MPO) in leukocytes was observed with laser confocal microscopy. The expression of phosphorylated extracellular signal-regulated kinase (phospho-extracellular signal kinase 1/2, p-ERK1 / 2) in leukocytes was detected by western blot. Results There was no significant change in the level of TNF-α in peripheral blood leukocytes of Mice treated with Hcy, while the level of MCP-1 was significantly increased, while the secretion of MPO increased and the expression of p-ERK1 / 2 increased significantly. CA inhibited Hcy-induced secretion of MCP-1 and MPO, a mouse peripheral leukocyte inflammatory mediator, and an increase in p-ERK1 / 2 protein expression. Conclusion CA can inhibit the release of MCP-1 and MPO from Hcy-induced mouse peripheral blood leukocytes, which may be related to the inhibition of p-ERK1 / 2 expression.