混合谱系激酶域样假激酶参与小鼠高原脑水肿的初步研究  

Preliminary study on the involvement of mixed lineage kinase domain-like pseudokinase in high-altitude cerebral edema in mice

作　　者：孟欣宇 孟瀚 毛竹青 王砚锦 谈妍辰 王亚周 崔毅 杨清湖[1] MENG Xinyu;MENG Han;MAO Zhuqing;WANG Yanjin;TAN Yanchen;WANG Yazhou;CUI Yi;YANG Qinghu(School of Life Sciences&Research Center for Natural Peptide Drugs,Shaanxi Engineering&Technological Research Center for Conversation&Utilization of Regional Biological Resources,Yan'an University,Yan'an 716099,China;Department of Neurobiology,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;No.2 Cadet Regiment,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;No.5 Cadet Regiment,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;Dalian Rehabilitation and Recuperation Center of Joint Logistics Support Force of PLA,Dalian 116013,China)

机构地区：[1]延安大学生命科学学院多态资源药物研究中心,陕西省区域生物资源保育与利用工程技术研究中心,陕西延安716099 [2]空军军医大学基础医学院神经生物学教研室,陕西西安710032 [3]空军军医大学基础医学院学员二大队,陕西西安710032 [4]空军军医大学基础医学院学员五大队,陕西西安710032 [5]联勤保障部队大连康复疗养中心,辽宁大连116013

出　　处：《空军军医大学学报》2025年第4期521-525,共5页Journal of Air Force Medical University

基　　金：国家自然科学基金面上项目(82171346)。

摘　　要：目的探索程序性坏死关键分子混合谱系激酶域样假激酶(MLKL)在小鼠高原脑水肿(HACE)中的作用。方法使用成年雄性C57BL/6 WT小鼠和MLKL^(-/-)小鼠,将小鼠分为3组,分别为对照组、WT HACE组和MLKL^(-/-)HACE组,每组8只。将小鼠放进小型高低压氧舱,调整海拔至5500 m,暴露72 h,建立HACE模型。使用Western blotting检测MLKL蛋白的表达。采用脑含水量指数评估脑水肿发生的程度。采用伊文思蓝检测血脑屏障开放状态。通过免疫荧光染色检测MLKL及水通道蛋白4(AQP4)在血管的分布,免疫球蛋白IgG的渗透。应用新物体识别实验分析小鼠认知功能。结果小鼠在5500 m下暴露72 h可导致血脑屏障破坏,脑含水量升高(P<0.01),新物体识别能力下降(P<0.01),MLKL及pMLKL表达升高(P<0.05),pMLKL与AQP4、CD31免疫荧光共标结果成点状聚集体。MLKL^(-/-)小鼠在同等刺激下呈现出较低程度的HACE,表现为MLKL^(-/-)HACE组小鼠较WT HACE组小鼠脑水含量显著减少(P<0.05),血脑屏障开放程度降低,免疫球蛋白从血管中的渗出减少,新物体识别功能改善(P<0.05)。结论程序性坏死关键分子MLKL参与HACE的发生与发展,敲除MLKL可减轻HACE。Objective To explore the role of mixed lineage kinase domain-like pseudokinase(MLKL),a key molecule in necroptosis,in high-attitude cerebral edema(HACE)in mice.Methods Adult male C57BL/6 wild-type mice and MLKL knockout(MLKL^(-/-))mice were divided into three groups:control group,WT HACE group and MLKL^(-/-)HACE group,with 8 mice in each group.Mice were placed into a small high-altitude hypoxia chamber.The altitude was adjusted to 5500 m,and mice were exposed for 72 h to establish HACE model.The expression of MLKL was detected by Western blotting.Brain water content was adopted to assess the degree of cerebral edema.Evans blue was used to evaluate brain-blood barrier permeability.Immunofluorescence staining was used to examine the distribution of MLKL and aquaporin-4(AQP4)in blood vessels and the effusion of IgG.Novel object recognition test was used to analyze the cognitive function of mice.Results Exposure to 5500 m for 72 h led to blood-brain barrier disruption,increased brain water content(P<0.01),decreased novel object recognition ability(P<0.01),increased expression of MLKL and pMLKL(P<0.05),and dot aggregates in immunofluorescence co-labeling of pMLKL with AQP4 and CD31 in mice.MLKL^(-/-)mice showed a lower degree of HACE under the same stimulation,manifested as a significant reduction of brain water content in MLKL^(-/-)HACE group as compared to WT HACE group(P<0.05),a decrease of blood-brain barrier opening,a reduction of immunoglobulin leakage from blood vessels,and an improvement of novel object recognition.Conclusion MLKL,a key molecule of necroptosis,is involved in the occurrence and development of HACE,and knockout of MLKL can alleviate HACE.

关 键 词：高原脑水肿 MLKL 血脑屏障 程序性细胞坏死 细胞毒性水肿 小鼠 

分 类 号：R594.3[医药卫生—内科学]