Ghrelin对棕榈酸诱导的THP-1巨噬细胞上TLR4/NF-κB信号通路活化的作用  

Effect of ghrelin on the activation of TLR4/NF-κB signaling pathway in THP-1 macrophages induced by palmitic acid

作　　者：李欣颖[1] 刘石平[1] 姚岚[1] 肖扬[1] 李卉[1] 周智广[1] 

机构地区：[1]中南大学湘雅二医院代谢内分泌研究所,长沙410011

出　　处：《中国糖尿病杂志》2012年第9期692-696,共5页Chinese Journal of Diabetes

基　　金：湖南省科技厅项目(2011hj3019);湖南省自然科学基金资助项目(11JJ7005)

摘　　要：目的探讨酰基化Ghrelin对棕榈酸(PA)诱导的人源单核巨噬细胞(THP-1巨噬细胞)上TLR4-NF-κB信号通路的作用。方法用不同浓度的PA孵育THP-1巨噬细胞株12h;用不同浓度的酰基化Ghrelin孵育THP-1巨噬细胞株4h后,再加入PA(200μmol/L)孵育12h。用实时定量PCR检测THP-1巨噬细胞上TLR4mRNA的表达水平;用Western印记检测TLR4蛋白和细胞内NF-κB p65磷酸化蛋白水平;用ELISA方法检测细胞上清液中IL-1β和TNF-α的浓度。结果与对照组比较,PA呈剂量依赖性增强THP-1巨噬细胞TLR4mRNA和蛋白的表达水平,以及NF-κB p65磷酸化蛋白水平和分泌TNF-α、IL-1β的能力(P均<0.05)。与PA(200μmol/L)组比较,酰基化Ghrelin呈剂量依赖性抑制THP-1巨噬细胞TLR4mRNA(P均<0.05)、TLR4蛋白和NF-κB p65磷酸化蛋白的水平(P均<0.01),以及降低THP-1巨噬细胞分泌TNF-α水平(P均<0.05)和IL-1β的水平(P均<0.01)。结论PA可剂量依赖性诱导THP-1巨噬细胞TLR4/NF-κB信号通路的活化;酰基化Ghrelin可剂量依赖性抑制PA诱导的THP-1巨噬细胞TLR4/NF-κB信号通路的活化。Objective To investigate the effect of acylated ghrelin activating TLR4/NF-κB signaling pathway in human monocyte-derived macrophages（THP-1） induced by palmitic acid.Methods THP-1 macrophage strains were cultured by the palmitic acid in various concentrations for 12 h,treated by acylated ghrelin in different concentrations for 4 h,and then cultured with the palmitic acid 200 μmol/L for 12 h again.The expression of TLR4 mRNA on the THP-1 macrophages was determined by quantitative real-time PCR.The levels of TLR4 protein and NF-κB p65 phosphorylation protein were measured by Western blot,and the concentrations of IL-1β and TNF-α in the cell supernatant were detected by ELISA.Results Compared with the control group,the palmitic acid increased the levels of TLR4 mRNA and protein,the NF-κB p65 phosphorylation protein,and the secretion of TNF-α and IL-1β of the THP-1 macrophages in a dose-dependent fashion（all P〈0.05）.Compared to the palmitic acid 200 μmol/L group,the acylated ghrelin inhibited the levels of TLR4 mRNA（all P〈0.05）,TLR4 protein,and the NF-κB p65 phosphorylation protein（all P〈0.01）,and decreased the secretion of TNF-α（all P〈0.05） and IL-1β（all P〈0.01） of the THP-1 macrophages in a dose-dependent fashion.Conclusions Palmitic acid can induce the activation of TLR4/NF-κB signaling pathway in THP-1 macrophages in a dose-dependent fashion.Acylated ghrelin can inhibit the activation of TLR4/NF-κB signaling pathway in THP-1 macrophages induced by palmitic acid in a dose-dependent fashion.

关 键 词：GHRELIN 棕榈酸 炎症 TOLL样受体 

分 类 号：R589.2[医药卫生—内分泌]