β-arrestin2基因敲除对小鼠腹腔巨噬细胞功能的影响  

Effect ofβ-arrestin2 deficiency on the function of mouse peritoneal macrophages

作　　者：陈婷婷 单杉 李南 汪子颖 杞萌 张胜男 胡姗姗[1] 魏伟[1] 孙妩弋[1] CHEN Tingting;SHAN Shan;LI Nan;WANG Ziying;QI Meng;ZHANG Shengnan;HU Shanshan;WEI Wei;SUN Wuyi(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anhui-inflammatory and Immune Medicine,Hefei 230032,China)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,抗炎免疫药物安徽省协同创新中心,合肥230032

出　　处：《中国实验动物学报》2022年第1期40-46,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(81770605,81300332);安徽医科大学科研水平提升计划(2021xkjT016)。

摘　　要：目的探讨β-arrestin2基因敲除对小鼠腹腔巨噬细胞(peritoneal macrophage,pMφ)功能的影响。方法分别分离野生型(wild type,WT)和β-arrestin2基因敲除(β-arrestin2-/-)小鼠的pMφ,Transwell法检测β-arrestin2基因敲除对pMφ迁移的影响;中性红吞噬实验检测缺失β-arrestin2基因的pMφ吞噬功能的变化;流式细胞术检测pMφ表面分子CD86的变化情况;ELISA法检测β-arrestin2基因敲除对pMφ炎性因子产生的影响;Western Blot法检测β-arrestin2、JAK1/STAT1通路相关蛋白的表达。结果与WT组相比,敲除β-arrestin2明显降低了pMφ的迁移能力,增强了pMφ的吞噬功能,且上调pMφ表面CD86分子的表达,同时pMφ分泌炎性因子IL-1β、TNF-α、IL-6的水平也明显升高;Western Blot检测结果表明,缺失β-arrestin2的pMφ中p-JAK1、p-STAT1的表达明显上调。结论β-arrestin2对小鼠pMφ的迁移、吞噬、极化等功能具有调节作用,其机制可能与调控JAK1/STAT1通路有关。Objective To investigate the effect ofβ-arrestin2 deficiency on the function of mouse peritoneal macrophages(pMφ).Methods The primary mouse pMφwere isolated from wild type mice andβ-arrestin2 gene knockout mice.The migration of pMφwas measured by Transwell assay.Changes in the phagocytosis of pMφafterβ-arrestin2 gene knockout were detected by the neutral red phagocytosis test.The expression of CD86 on pMφwas analyzed by flow cytometry.The levels of interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)in pMφwere detected by ELISA.Protein expression levels ofβ-arrestin2,JAK1,p-JAK1,STAT1 and p-STAT1 were detected by Western Blot.Results Compared with the control group,β-arrestin2 knockout significantly inhibited the migration ability of pMφ,and significantly enhanced the phagocytosis of pMφand the expression of CD86.The production of IL-1β,IL-6 and TNF-αwas significantly increased in pMφafterβ-arrestin2 depletion.The result of Western Blot showed thatβ-arrestin2 deficiency significantly up-regulated the levels of p-JAK1 and p-STAT1 in pMφ.Conclusions These result indicated thatβ-arrestin2 plays an important role in regulating the migration,phagocytosis and polarization of pMφ,and these effects appear to be mediated by the JAK1/STAT1 signaling pathway.

关 键 词：β-arrestin2 腹腔巨噬细胞 炎症因子 JAK1 STAT1 

分 类 号：Q95-33[生物学—动物学]