【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-kB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12 h;用不同浓度的酰基化ghrelin孵育THP-1巨噬细胞株4 h后，再加入棕榈酸(200μmol/L)孵育12h。用实时定量PCR检测THP-1巨噬细胞上TLR4 mRNA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TLR4 mRNA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(均P<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TLR4 mRNA(均P<0.05)、TLR4蛋白和NF-κB p65磷酸化蛋白的水平(均P<0.01)，以及降低THP-1巨噬细胞分泌 TNF-α水平(均P<0.05)和 IL-1β的水平(均P <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。

　　【关键词】 Ghrelin;棕榈酸;炎症;Toll样受体

　　Effect of ghrelin on the activation of TLR4/NF-κB signaling pathway in THP-1 macrophages induced by palmitic acid LI Xin-ying, LIU Shi-ping，YAO Lan，et al. Institute of Metabolism and Endocrinology, the Second Xiangya Hospital, Central South University, Changsha 410011, China

　　Corresponding author: LIU Shi-ping, Email: shipingliu119@126.com

　　【Abstract】 Objective To investigate the effect of acylated ghrelin on the activation of TLR4/NF-κB signaling pathway induced by palmitic acid in human monocyte-derived macrophages (THP-1). Methods THP-1 macrophage strains were cultured by the palmitic acid in various concentrations for 12h, treated by acylated ghrelin in different concentrations for 4h, and then cultured with the palmitic acid 200μmol/L for 12h 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 blotting, and the concentrations of IL-1β and TNF-α in the cell supernate 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-1βof the THP-1 macrophages in a dose-dependent fashion (all P<0.05). Compared to the palmitic acid 200μmol/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), 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.

　　【Key Words】 Ghrelin; Palmitic acid; Inflammation; Toll-like receptor