鸢尾素通过调控内质网应激抑制脂多糖诱导的THP-1巨噬细胞炎症反应  

Irisin inhibits lipopolysaccharide-induced THP-1 macrophage inflammation by regulating endoplasmic reticulum stress

作　　者：符宗冬 李鸽 林雪娟 陈丽君 张大启 李其富[1,2] 廖小平 赵文杰[1] FU Zong-dong;LI Ge;LIN Xue-Juan(Department of Neurology,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan,China)

机构地区：[1]海南医学院第一附属医院神经内科,海南海口570102 [2]海南省热带脑科学研究与转化重点实验室

出　　处：《中国老年学杂志》2023年第17期4214-4218,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金(82260250);海南省自然科学基金高层次人才项目(822RC823)。

摘　　要：目的探讨鸢尾素(Irisin)在脂多糖(LPS)诱导的人髓系白血病单核细胞(THP)-1巨噬细胞炎症中的作用及机制。方法采用佛波酯诱导THP-1单核细胞分化为巨噬细胞,LPS刺激构建THP-1巨噬细胞炎症模型,分为正常对照组、不同浓度Irisin组、LPS组、LPS+不同浓度Irisin组、LPS+Irisin+衣霉素(TM)组。采用酶联免疫吸附试验检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β和IL-6水平。采用线粒体膜电位(MMP)试剂盒测定MMP。采用活性氧(ROS)试剂盒测定ROS水平。采用Western印迹检测肌醇需要酶(IRE)1α、双链RNA激活的蛋白激酶类内质网激酶(PERK)、免疫球蛋白重链结合蛋白(BIP)、phospho-核转录因子(NF)-κB p65和NF-κB p65的表达。结果与正常对照组相比,LPS诱导的THP-1巨噬细胞上清液中TNF-α、IL-6和IL-1β水平明显升高(均P<0.05);200 ng/ml Irisin处理后,LPS诱导的THP-1巨噬细胞TNF-α、IL-6和IL-1β分泌显著下降(均P<0.05);200 ng/ml Irisin和TM共同处理LPS诱导的THP-1巨噬细胞后,TNF-α、IL-6和IL-1β水平又显著升高(均P<0.05)。与正常对照组相比,LPS组ROS水平明显升高,MMP明显下降(均P<0.05),而Irisin则可显著抑制LPS诱导的THP1巨噬细胞ROS水平升高和MMP下降(均P<0.05)。与LPS组相比,LPS+Irisin组BIP、PERK和IRE1表达水平及phospho-NF-κB p65/NF-κB p65比值均明显降低(P<0.05),而与LPS+Irisin组相比,LPS+Irisin+TM组以上指标均明显升高(P<0.05)。结论Irisin可通过抑制内质网应激和NF-κB信号通路,改善氧化应激和线粒体功能,进而抑制LPS诱导的THP-1巨噬细胞炎症反应。Objective To explore the role of Irisin in lipopolysaccharide(LPS)-induced inflammation in human myeloid leukemia mononuclear cell(THP)-1 macrophages and the mechanisms.Methods The differentiation of THP-1 monocytes into macrophages was induced by phoboate,and the inflammation model of THP-1 macrophages was constructed by LPS stimulation,which was divided into normal control group,different concentrations of Irisin group,LPS group,LPS+different concentrations of Irisin group and LPS+Irisin+tunicamycin(TM)group.Tumor necrosis factor(TNF)-α,interleukin(IL)-1βand IL-6 levels were measured by enzyme-linked immunosorbent.Mitochondrial membrane potential(MMP)was determined by MMP assay kit.The levels of reactive oxygen species(ROS)were measured by ROS assay kit.The expressions of inositol-requiring enzyme(IRE)1α,double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase(PERK),immunoglobulin heavy chain binding protein(BIP),phospho-nuclear factor(NF)-κB p65 and NF-κB p65 were detected by Western blotting.Results Compared to the normal control group,the levels of TNF-α,IL-6 and IL-1βwere significantly increased in the supernatant of LPS-induced THP-1 macrophages(all P<0.05);after 200 ng/ml Irisin treatment,the secretion of TNF-α,IL-6 and IL-1βwere significantly decreased in LPS-induced THP-1 macrophages(all P<0.05);and the levels of TNF-α,IL-6 and IL-1βin LPS-induced THP-1 macrophage were significantly increased after co-treatment with 200 ng/ml Irisin and TM(all P<0.05).Compared with the normal control group,the level of ROS was significantly increased and MMP was significantly decreased in LPS group(all P<0.05),while Irisin could significantly inhibit the increase of ROS level and the decrease of MMP in THP1 macrophages induced by LPS(all P<0.05).Compared with the LPS group,the LPS+Irisin group showed a significant decrease in the expression levels of BIP,PERK,IRE1 and the ratio of phosphor-NF-κB p65 to NF-κB p65(P<0.05),and compared with the LPS+Irisin group,the above indexes in LPS+Ir

关 键 词：鸢尾素 脂多糖 人髓系白血病单核细胞(THP)-1巨噬细胞 

分 类 号：R741[医药卫生—神经病学与精神病学]