咖啡酸苯乙酯对ox-LDL诱导的THP-1源性巨噬细胞炎症因子分泌的抑制作用

Caffeic Acid Phenethyl Ester Suppresses Inflammatory Cytokines Secretion in ox-LDL Treated THP-1 Macrophages

目的研究咖啡酸苯乙酯(CAPE)对ox-LDL诱导的THP-1源性巨噬细胞炎症因子分泌的影响及其可能机制。方法用不同浓度CAPE预处理THP-1源性巨噬细胞2 h,再用40 mg/L ox-LDL处理24 h,ELISA检测炎症因子TNF-α,IL-6以及MCP-1的分泌情况;用40 mg/L ox-LDL分别处理THP-1源性巨噬细胞不同时间,Westernblot检测细胞COX-2蛋白表达的情况;最后,采用Western blot分析CAPE对ox-LDL诱导的THP-1源性巨噬细胞COX-2蛋白表达、IκB-α降解及其NF-κB核转位的影响。结果 40 mg/L ox-LDL可诱导THP-1源性巨噬细胞TNF-α,IL-6以及MCP-1分泌增多,而CAPE明显抑制了ox-LDL诱导的细胞炎症因子分泌,呈浓度依赖性(P<0.05);随着ox-LDL处理时间的延长,THP-1源性巨噬细胞COX-2蛋白表达逐渐增高,以24 h最为明显(P<0.05),而CAPE能抑制ox-LDL诱导的THP-1源性巨噬细胞COX-2蛋白表达上调,并可抑制ox-LDL诱导的THP-1源性巨噬细胞IκB-α降解及其NF-κB核转位。结论 CAPE对ox-LDL诱导的巨噬细胞炎症因子分泌应具有抑制作用,作用机制可能与其抑制NF-κB激活,下调COX-2蛋白表达有关。

Aim To investigate the effect and underlying mechanisms of Caffeic acid phenethyl ester（CAPE） on inflammatory cytokines secretion in oxidized low density lipoprotein （ox-LDL）-stimulated THP-1 macrophages, netla- ads The THP-! macrophages were preincubated with CAPE at different concentrations for 2 h, and then treated with ox- LDL（40 mg/L） for 24 h. The secret levels of TNF-o~, IL-6 and MCP-1 in cells were determined by ELISA. After trea- ted with 40 mg/L of ox-LDL for the indicated times, the expression of COX-2 in THP-1 macrophages was measured by Western blot. Subsequently, THP-1 cells were pretreated with indicated concentrations of CAPE, and then incubated with or without 40 mg/L of ox-LDL for 1 h. The expressions of COX-2, IKB-α and nuclear NF-KB were measured by Western blot. Results CAPE suppresses the ox-LDL-induced up-regulation of TNF-α, IL-6 and MCP-1 in THP-1 macropha- ges. The expression of COX-2 was markedly increased by ox-LDL, which, however, could be significantly attenuated by CAPE. CAPE suppresses ox-LDL induced IKB-α degradation and NF-KB nuclear translocation in THP-1 macrophages. Conclusion CAPE suppresses inflammatory cytokines secretion in ox-LDL treated THP-1 macrophages through inhibition of NF-kB activation and COX-2 expression.