膜联蛋白A1的N末端肽Ac2-26对高糖刺激的小鼠巨噬细胞极化的影响  

Effect of annexin A1 N-terminal derived peptide Ac2-26 on high glucosestimulated mouse macrophage polarization

作　　者：夏崇建 黄俊杰 张煜 卓嘉英 吴娟娟 陈刚 范琰琰 XIA Chong-jian;HUANG Jun-jie;ZHANG yu;ZHUO Jia-ying;WU Juan-juan;CHENGang;FAN Yan-yan(School of Basic Medical Science,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学基础医学院,浙江温州325035

出　　处：《中国病理生理杂志》2022年第11期1998-2004,共7页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.LY20H150006);国家自然科学基金资助项目(No.81301640)。

摘　　要：目的:探讨膜联蛋白A1(ANXA1)的N末端肽Ac2-26对高糖刺激的小鼠巨噬细胞极化的影响及其机制。方法:体外培养小鼠骨髓来源的巨噬细胞(BMDM),将其分为对照组、高糖组、高糖+Ac2-26组、高糖+Ac2-26+甲酰肽受体2(FPR2)拮抗剂WRW4组和高糖+Ac2-26+磷脂酰肌醇3-激酶(PI3K)抑制剂wortmannin组。ELISA检测BMDM上清液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10和转化生长因子β1(TGF-β1)的分泌;Western blot检测BMDM中诱导型一氧化氮合酶(iNOS;M1型极化标志物)、CD206(M2型极化标志物)及ANXA1的表达,并检测PI3K和蛋白激酶B(PKB/AKT)的表达及磷酸化水平。结果:高糖刺激可以显著上调小鼠BMDM的iNOS表达及促炎因子(TNF-α和IL-6)产生(P<0.01),下调CD206表达和抗炎因子(IL-10)产生(P<0.05),并抑制细胞内PI3K/AKT的磷酸化(P<0.05);使用Ac2-26对高糖刺激的BMDM进行干预,能够显著抑制iNOS表达及促炎因子(TNF-α和IL-6)产生(P<0.05),并上调CD206表达、IL-10产生及PI3K/AKT的磷酸化水平(P<0.05);而WRW4和wortmannin能够显著抑制Ac2-26的效应(P<0.05)。结论:高糖刺激可以诱导小鼠BMDM向促炎的M1型极化,而Ac2-26可以抑制M1型极化,并促进BMDM向抗炎的M2型极化;Ac2-26的调节机制可能涉及FPR2/PI3K/AKT信号通路的激活。AIM:To investigate the effect of annexin A1(ANXA1)N-terminal derived peptide Ac2-26 on high glucose(HG)-stimulated mouse macrophage polarization,and to explore its possible mechanisms.METHODS:Bone marrow-derived macrophages(BMDMs)were isolated from mice.Then,BMDMs were divided into the following groups:control group,high glucose(HG)group,HG+Ac2-26 group,HG+Ac2-26+formyl peptide receptor 2(FPR2)antagonist WRW4 group and HG+Ac2-26+phosphatidylinositol 3-kinase(PI3K)inhibitor wortmannin group.ELISA kits were used to determine the secretion of tumor necrosis factorα(TNF-α),interleukin 6(IL-6),IL-10 and transforming growth factorβ1(TGF-β1)in BMDM from each group.Western blot was applied to detect the protein levels of inducible nitric oxide synthase(iNOS;M1 polarization marker),CD206(M2 polarization marker),PI3K,phosphorylated PI3K,protein kinase B(Akt)and phosphorylated Akt.RESULTS:High glucose stimulation significantly increased iNOS expression and pro-inflammatory cytokine(TNF-αand IL-6)production(P<0.01),decreased CD206 expression and IL-10 secretion(P<0.05),and inhibited the phosphorylation of PI3K/Akt(P<0.05)in mouse BMDM.Ac2-26 treatment in high glucose-stimulated BMDM suppressed iNOS expression and pro-inflammatory cytokine(TNF-αand IL-6)production(P<0.05),and up-regulated CD206 expression,IL-10 production and the phosphorylation level of PI3K/Akt(P<0.05).Moreover,FPR2 antagonist(WRW4)and PI3K inhibitor(wortmannin)significantly inhibited the effects of Ac2-26 on high glucose-stimulated BMDM(P<0.05).CONCLUSION:High glucose stimulation can induce the pro-inflammatory M1 polarization of mouse BMDM,while Ac2-26 can inhibit the M1 polarization of BMDM and promote the anti-inflammatory M2 polarization.The regulatory mechanism of Ac2-26 may involve the activation of FPR2/PI3K/Akt signaling pathway.

关 键 词：糖尿病 巨噬细胞 极化 膜联蛋白A1 Ac2-26 FPR2/PI3K/AKT信号通路 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R587.1[医药卫生—基础医学]