胫骨骨折小鼠海马巨噬细胞极化与围术期神经认知障碍的关系  

Relationship between hippocampal macrophage polarization and perioperative neurocognitive disorders in mice with tibial fracture

作　　者：林亚男 李燕玲[1] 张浩然 王彬[1] 林旭[1] 宋巧玲 张晓楠 赵俊 毕燕琳[1] Lin Yanan;Li Yanling;Zhang Haoran;Wang Bin;Lin Xu;Song Qiaoling;Zhang Xiaonan;Zhao Jun;Bi Yanlin(Department of Anesthesiology,Qingdao Hospital,University of Health and Rehabilitation Sciences(Qingdao Municipal Hospital),Qingdao 266071,China;Department of Anesthesiology,Weifang Medical University,Weifang 261053,China;Innovation Center for Marine Drug Screening and Evaluation Qingdao National Laboratory for Marine Science and Technology,Key Laboratory of Marine Drugs,Qingdao 266100,China)

机构地区：[1]康复大学青岛医院(青岛市市立医院)东院麻醉科,青岛266071 [2]潍坊医学院麻醉学院,潍坊261053 [3]青岛海洋科学与技术试点国家实验室,海洋创新药物筛选与评价平台,青岛266100

出　　处：《中华麻醉学杂志》2023年第5期564-569,共6页Chinese Journal of Anesthesiology

摘　　要：目的:评价胫骨骨折小鼠海马巨噬细胞极化与围术期神经认知障碍(PND)的关系。方法:清洁级健康雄性C57/BL6小鼠45只,5~7月龄,采用随机数字表法分为3组(n=15):对照组(C组)、麻醉组(A组)和麻醉手术组(AS组)。C组不做任何处理。A组吸入2%异氟烷麻醉15 min。AS组吸入2%异氟烷麻醉下行胫骨骨折髓内钉固定术。于麻醉前/术前1 d、麻醉后/术后1、3、7 d时进行Morris水迷宫实验和旷场实验。各时点行为学实验结束后随机取5只小鼠,处死后取海马组织,采用qRT-PCR法检测海马TNF-α、IL-1β、IL-6、趋化因子2(CCL2)、CCL5、CCL8、一氧化氮合酶(iNOS)及精氨酸酶-1(Arg-1)的mRNA表达,采用Western blot法检测海马组织iNOS和Arg-1的表达水平,采用免疫荧光染色法测定海马区CD11b^(+)、CD45^(+)、CD86^(+)和CD206^(+)细胞百分比。结果:与C组相比,AS组术后逃避潜伏期延长,穿越平台次数减少,海马TNF-αmRNA、IL-6 mRNA、CCL5 mRNA、CCL8 mRNA、iNOS及其mRNA表达上调,Arg-1及其mRNA表达下调,海马CD11b^(+)CD45^(+)细胞百分比、CD11b^(+)CD86^(+)细胞百分比升高,CD11b^(+)CD206^(+)细胞百分比降低(P<0.05),A组上述指标差异无统计学意义(P>0.05);与A组相比,AS组术后逃避潜伏期延长,穿越平台次数减少,iNOS mRNA表达上调,CD11b^(+)CD45^(+)细胞百分比、CD11b^(+)CD86^(+)细胞百分比升高,CD11b^(+)CD206^(+)细胞百分比降低(P<0.05)。结论:胫骨骨折小鼠PND的发生可能与海马M1型巨噬细胞极化增加,且M2型巨噬细胞极化减少,进而引起中枢炎症反应有关。Objective To evaluate the relationship between hippocampal macrophage polarization and perioperative neurocognitive disorders in mice with tibial fractures.MethodsForty-five clean-grade healthy male C57/BL6 mice,aged 5-7 months,were divided into 3 groups(n=15 each)using the random number table method:control group(group C),anesthesia group(group A)and anesthesia surgery group(group AS).Group C received no treatment.Group A was anesthetized with isoflurane inhaled for 15 min.In AS group,intramedullary nail fixation of tibial fracture was performed under anesthesia through inhalation of 2%isoflurane.Morris water maze test and open field test were performed before anesthesia/on 1 day before surgery and after anesthesia/on 1,3 and 7 days after operation.Five mice were randomly selected after the behavioral experiments were completed at each time point,and hippocampal tissues were taken after the animals were sacrificed for determination of the expression of tumor necrosis factor-alpha(TNF-α),interleukin-1 beta(IL-1β),IL-6,chemokine(c-c motif)ligand 12(CCL2),CCL5,CCL8,iNOS and Arg-1 mRNA(by quantitative real-time polymerase chain reaction),expression of iNOS and Arg-1 proteins(by Western blot),and percentage of CD11b,CD45,CD86 and CD206 cells in hippcampal area(by immunofluorescence staining).ResultsCompared with group C,the escape latency was significantly prolonged after operation,the number of crossing the platform was reduced after operation,the expression of TNF-ɑ,IL-6,CCL5 and CCL8 mRNA and iNOS protein and mRNA was up-regulated,the expression of Arg-1 protein and mRNA was down-regulated,the percentages of CD11b^(+)CD45^(+)cells and CD11b^(+)CD86^(+)cells in the hippocampus were increased,and the percentages of CD11b^(+)CD206^(+)cells were decreased in AS group(P<0.05),and no significant change was found in the parameters mentioned above in group A(P>0.05).Compared with group A,the escape latency was significantly prolonged after surgery,the frequency of crossing the platform was reduced after surgery,the exp

关 键 词：巨噬细胞 神经认知障碍 骨折固定术 

分 类 号：R614[医药卫生—麻醉学]