术后谵妄小鼠外周血源性外泌体对BV2小胶质细胞的影响  

Effects of Peripheral Blood-derived Exosomes on BV2 Microglia Cells in Mice with Postoperative Delirium

作　　者：汤莎灵 张芸 冯蓓 杨晓霞[1] 高宇博[1] TANG Shaling;ZHANG Yun;FENG Bei;YANG Xiaoxia;GAO Yubo(Department of Anesthesia and Perioperative Medicine,General Hospital of Ningxia Medical University,First Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院麻醉与围术期医学科,宁夏医科大学第一临床医学院,银川750004

出　　处：《宁夏医科大学学报》2024年第4期360-365,共6页Journal of Ningxia Medical University

基　　金：宁夏留学回国人员创新创业项目。

摘　　要：目的 探讨术后谵妄(postoperative delirium,POD)小鼠外周血源性外泌体对BV2小胶质细胞的影响。方法 选择12~14月龄的C57BL/6J雄性小鼠16只,随机分为空白对照组(Con组)和POD组,采用莫里斯水迷宫和Y迷宫实验评价POD模型是否建立成功,随后采用纳米颗粒跟踪分析、透射电子显微镜和Western blot法对血清中的外泌体进行鉴定,用PKH26染料标记外泌体。将BV2细胞随机分为对照组(Nor组)、正常小鼠外周血源性外泌体组(Con-Exo组)和麻醉/手术小鼠外周血源性外泌体组(POD-Exo组),共孵育24 h后,采用CCK-8法检测各组细胞的活性,RT-qPCR和Western blot检测IL-1β、IL-6和TNF-α的相对mRNA和蛋白的表达。结果 与Con组相比,POD组小鼠的认知能力下降(P<0.05);共聚焦显微镜下观察到BV2细胞能够内吞被PKH26染料标记的外泌体;与Nor组和Con-Exo组比较,POD-Exo组BV2细胞活性下降(P<0.001),促炎因子IL-1β、IL-6和TNT-α的mRNA和蛋白表达均上升(P均<0.001)。结论 POD小鼠外周血源性Exo可诱导BV2细胞炎症反应。Objective To explore the effects of peripheral blood-derived exosomes on BV2 microglia cells in mice with postoperative delirium(POD).Methods Sixteen C57BL/6J male mice aged 12-14 months were used and randomly divided into control group(Con group)and postoperative delirium(POD)group.Morris water maze and Y maze experiments were used to evaluate whether the POD model was successfully established,and then nanoparticle tracking analysis,transmission electron microscopy,and western blot were used to identify exosomes in serum,and exosomes were labeled with PKH26 dye.The BV2 cells were randomly divided into three groups:normal group(Nor group),peripheral blood-derived exosomes from normal mice(Con-Exo group),and peripheral blood-derived exosomes from anesthetized/surgical mice(POD-Exo group),and were incubated for a total of 24 hours.Then the activity of the cells in each group was detected by the CCK-8 assay,and the IL-1β,IL-6,and TNF-αrelative mRNA and protein expression was detected by RT-qPCR and Western blot.Results Compared with the Con group,the cognitive ability of mice in the POD group was decreased(P<0.05).The ability of BV2 cells to endocytose exosomes labeled with PKH26 dye was observed under confocal microscopy.The activity of BV2 cells in the POD-Exo group was significantly decreased(P<0.001)compared with that of the Nor and Con-Exo groups and relative mRNA and protein expressions of the pro-inflammatory factors IL-1β,IL-6 and TNF-αincreased(P all<0.001).Conclusion Peripheral blood-derived exosomes in mice with POD can induce inflammatory responses in BV2 cells.

关 键 词：术后谵妄 BV2细胞 外泌体 炎性因子 

分 类 号：R619[医药卫生—外科学]