紫檀芪对棕榈酸诱导巨噬细胞炎症因子分泌的影响

Effect of pterostilbene on palmitic acid-induced secretion of inflammatory factors in macrophages

目的:研究紫檀芪(pterostilbene,PTE)对棕榈酸(palmitic acid,PA)诱导巨噬细胞炎症因子分泌的影响及可能机制。方法:采用MTT法检测PTE对巨噬细胞RAW264.7活性的影响;先用不同浓度的PTE预处理RAW264.7,后用PA处理,ELISA法检测巨噬细胞培养上清液中炎症因子IL-18、IL-1β和TNF-α水平,采用蛋白质印迹法检测巨噬细胞中NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing CARD,ASC)、半胱天冬酶1(caspase-1)、核转录因子-κB p65(NF-κB p65)、p-NF-κB p65的表达。结果:与对照组比较,0~200μmol/L浓度的PTE对巨噬细胞的活力无影响。50μmol/L和100μmol/L的PTE可显著抑制PA诱导的巨噬细胞炎症因子IL-18、IL-1β和TNF-α水平的升高,降低PA诱导巨噬细胞中NLRP3、ASC、caspase-1蛋白表达的升高。PTE可减少PA诱导胞质中p-IKK和p-NF-κB p65表达的升高,以及IκBα表达的降低。PTE还可降低PA诱导的p-NF-κB p65的核转位。结论:PTE可抑制PA诱导巨噬细胞炎症因子表达的升高,其机制可能与其抑制炎症小体活化的通路有关。

Objective:To study the effect of pterostilbene(PTE)on palmitic acid(PA)-induced inflammatory factor secretion in macrophages and its possible mechanism.Methods:MTT method was used to detect the effect of pterostilbene on the activity of RAW264.7 cells.RAW264.7 cells were pretreated with different concentrations of pterostilbene and then treated by PA.ELISA kit was used to detect the levels of IL-18,IL-1βand TNF-αin the supernatant of macrophages,and Western blotting was used to detect the expression of NLRP3,apoptosisassociated speck-like protein containing CARD(ASC),caspase-1,Nuclear factor ofκB p65(NF-κB p65),p-NF-κB p65 in macrophages.Results:Compared with the control group,MTT test results showed that PTE at a concentration of 0-200μmol/L had no effect on the activity of macrophages.PTE at 50 and 100μmol/L could significantly inhibit the PA-induced the expression of IL-18,IL-1βand TNF-α,and reduce PA-induced the expression of NLRP3,ASC,caspase-1 in macrophages.Meanwhile,PTE inhibited the expression of p-IKK and p-NF-κB p65,and reduced the decrease of the expression of IκBαin cytoplasm of PA-induced RAW264.7 cells.PTE can also reduce the nuclear translocation of p-NF-κB p65 induced by PA.Conclusion:Pterostilbene can inhibit PA-induced increase of inflammatory factor expression in macrophages,and its mechanism may be related to its inhibition of the pathway of inflammasomes.