银杏叶提取物对U937泡沫细胞IL-1β、TNF-α及IL-10表达的影响

摘要

本研究考察了氧化低密度脂蛋白(ox-LDL)刺激的U937细胞致炎细胞因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、抗炎细胞因子白细胞介素10(IL-10)及其受体(IL-10R)的蛋白及mRNA的表达,同时观察银杏叶提取物(GbE)对它们的作用.U937细胞用100 mg·L-1 ox-LDL刺激24 h形成泡沫细胞,同时分别加入不同浓度的GbE(0.1,1及10 μg·L-1)共孵育,采用酶联免疫吸附试验(ELISA)及逆转录聚合酶链式反应(RT-PCR)方法检测IL-1β、TNF-α、IL-10和IL-10R的蛋白或mRNA表达.U937泡沫细胞组IL-1β、TNF-α、IL-10和IL-10R的蛋白或mRNA的表达较对照组显著增加(P＜0.01).GbE组IL-1β及TNF-α的蛋白和mRNA的表达水平明显降低,IL-10的蛋白、IL-10和IL-10R的mRNA表达水平明显提高,与U937泡沫细胞组相比差异显著(P＜0.05,P＜0.01).GbE对U937泡沫细胞致炎细胞因子IL-1β及TNF-α表达的显著抑制作用,对抗炎细胞因子IL-10及其受体IL-10R表达的显著上调作用可能是其抗atherosclerosis(AS)的机制之一.%This study is to investigate the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in U937 foam cells and effects of Ginkgo biloba extract (GbE) on the cytokines.U937 cells were cultured with different concentrations of GbE (0.1,1,and 10 μg·L-1),and stimulated by 100 mg·L-1 oxidized low density lipoprotein (ox-LDL) for 24 h.The expressions of interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in culture solution were detected by enzyme-linked immunosorbant assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR).The results showed that incubated with 100 mg·L-1 ox-LDL for 24 h,the U937 cells became foam cells,the protein or mRNA expressions of IL-1β,TNF-α,IL-10,and its receptor IL-10R in U937 foam cells were higher markedly than those in normal U937 cells.When the cells were pretreated with GbE (0.1,1,and 10 μg·L-1),the increases of IL-1β and TNF-α in U937 foam cells were remarkably inhibited,but IL-10 expression increased greatly.Especially when cells were pretreated with 10 μg·L-1 GbE,the protein and mRNA expressions of IL-1β and TNF-α were markedly lower than those in U937 foam cells.The protein expression of IL-10 and mRNA expressions of IL-10 and its receptor IL-10R were markedly higher than those in U937 foam cells.GbE inhibited production of pro-inflammatory cytokines IL-1β and TNF-α,but up-regulated the production of anti-inflammatory cytokine IL-10 and its receptor IL-10R in U937 foam cells,which might be related with its anti-atherosclerotic actions.