基于单核巨噬细胞外泌体/microRNA-92a探讨AGEs对糖尿病内皮细胞损伤的作用机制  

To explore the mechanism of AGEs on diabetic endothelial cell damage based on monocyte-macrophage exosomes/microRNA-92a

作　　者：李雁[1] 张新菊[1] 刘武[1] 李金峰[1] 孙燕[1] 李辉[2] 程洁萍 Li Yan;Zhang Xinju;Liu Wu;Li Jinfeng;Sun Yan;Li Hui;Cheng Jieping(Dept of Endocrinology,Xinjiang Production and Construction Corps Hospital(The Second Affiliated Hospital of Shihezi University School of Medicine),Urumqi 830092;Administrative Organ,Xinjiang Production and Construction Corps Hospital(The Second Affiliated Hospital of Shihezi University School of Medicine),Urumqi 830092)

机构地区：[1]新疆生产建设兵团医院(石河子大学医学院第二附属医院)内分泌科,乌鲁木齐830092 [2]新疆生产建设兵团医院(石河子大学医学院第二附属医院)行政机关,乌鲁木齐830092

出　　处：《安徽医科大学学报》2023年第1期85-94,共10页Acta Universitatis Medicinalis Anhui

基　　金：兵团科技攻关项目(编号:2018AB024)。

摘　　要：目的基于单核巨噬细胞外泌体(Exos)/microRNA-92a(miR-92a)探讨晚期糖基化终产物(AGEs)对糖尿病内皮细胞损伤的作用机制。方法20只载脂蛋白E缺乏(ApoE^(-/-))小鼠随机均分为两组:损伤组和损伤+STZ组。损伤+STZ组建立链脲佐菌素(STZ)诱导的糖尿病模型。所有动物都接受了部分左颈动脉(PLCA)结扎手术。收集颈动脉,采用免疫组化检测M1巨噬细胞数量,并使用ELISA分析AGEs水平。用AGEs处理RAW264.7细胞的条件培养基(CM)或AGEs刺激巨噬细胞来源Exos处理微血管内皮细胞系bEnd.3细胞,然后评估细胞屏障功能和线粒体功能。结果糖尿病小鼠颈动脉粥样硬化组织和AGEs处理的RAW264.7细胞中M1巨噬细胞数量增加。CM或Exos在体外诱导了血管内皮细胞的屏障功能障碍、活性氧(ROS)积累和线粒体功能障碍。此外,生物信息学分析表明miR-92a在AGEs刺激巨噬细胞来源Exos中上调。实验上,Exos通过转移miR-29a参与了bEnd.3细胞中CM诱导的屏障功能障碍、ROS积累和线粒体功能障碍。最后,一系列拯救实验进一步证实Exos通过miR-92a调节血管内皮细胞的屏障功能障碍和线粒体功能。结论糖尿病ApoE^(-/-)小鼠AGEs表达和M1巨噬细胞数量增加,并且AGEs刺激巨噬细胞来源Exos通过体外递送miR-92a诱导bEnd.3细胞的屏障功能和线粒体功能障碍。Objective To explore the mechanism of advanced glycation end products(AGEs)on diabetic endothelial cell damage based on monocyte-macrophage exosomes(Exos)/microRNA-92a(miR-92a).Methods Twenty apolipoprotein E-deficient(ApoE^(-/-))mice were randomly divided into two groups:injury group(n=10)and injury+STZ group(n=10).The injury+STZ group established a diabetes model induced by streptozotocin(STZ).All animals underwent partial left carotid artery(PLCA)ligation.The carotid arteries were collected,the number of M1 macrophages was detected by immunohistochemistry,and the level of AGEs was analyzed by ELISA.Microvascular endothelial cell line bEnd.3 cells were treated with conditioned medium(CM)of AGEs treated RAW264.7 cells or Exos derived from RAW264.7,followed by evaluations of the cell barrier function and mitochondrial function.Results There was an increased number of M1 macrophages in carotid atherosclerotic tissues of diabetic mice and in AGEs treated RAW264.7 cells.CM or Exos significantly induced barrier dysfunction,reactive oxygen species(ROS)accumulation and mitochondrial dysfunction in vascular endothelial cellsin vitro.In addition,bioinformatics analysis showed that miR-92a was up-regulated in Exos derived from macrophages stimulated by AGEs.Experimentally,Exos participated in CM-induced barrier dysfunction,ROS accumulation and mitochondrial dysfunction in bEnd.3 cells by transferring miR-92a.Finally,a series of rescue experiments further confirmed that Exos regulated the barrier dysfunction and mitochondrial function in vascular endothelial cells through miR-92a.Conclusion The expression of AGEs and the number of M1 macrophages in diabetic ApoE^(-/-)mice increase,and AGEs stimulates Exos from macrophages could impair the barrier function and mitochondrial function in vascular endothelial cells by delivering miR-92a in vitro.

关 键 词：巨噬细胞 外泌体 microRNA-92a 晚期糖基化终产物 糖尿病 内皮细胞损伤 

分 类 号：R725.6[医药卫生—儿科]