抑制NKCC1/AQP4通路对改善高原脑水肿大鼠神经损伤的作用  

Effects of Inhibiting the NKCC1/AQP4 Pathway on Neurological Injury Improvement in a Rat Model of High-Altitude Cerebral Edema

作　　者：耿华丽 李百川 宋旭[3] 夏逸林 周向阳 高静 陈蕾[1,2] GENG Huali;LI Baichuan;SONG Xu;XIA Yilin;ZHOU Xiangyang;GAO Jing;CHEN Lei(High Altitude Medicine Key Laboratory of Sichuan Province,West China Hospital,Sichuan University,Chengdu 610041,China;Laboratory of Neurology and Comorbidities,West China Hospital,Sichuan University,Chengdu 610041,China;College of Life Sciences,Sichuan University,Chengdu 610065,China)

机构地区：[1]四川大学华西医院高原医学研究院,成都610041 [2]四川大学华西医院神经和共病研究室,成都610041 [3]四川大学生命科学学院,成都610065

出　　处：《四川大学学报(医学版)》2025年第1期156-165,共10页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅四川省中央引导地方科技发展专项(No.2023ZYD0075);四川大学华西医院横向合作项目(No.HX-H2307204)资助。

摘　　要：目的探究高原脑水肿(high-altitude cerebral edema,HACE)的发病机制,开发新型治疗策略。方法选取6周龄SD雄性大鼠,置于低压舱内,并选定7000 m海拔处理3 d为造模条件,通过测量脑含水量、血脑屏障(blood-brain barrier,BBB)破坏程度、脑组织尼式染色,评估高原脑水肿大鼠模型的建立是否成功。将实验动物分为4组,每组28只。空白对照组于常压常氧模拟500 m海拔3 d;模型组(HACE组)、布美他尼组(阳性对照组)、XH-6003药物组置于7000 m海拔,于尾静脉分别注射生理盐水、布美他尼和新型钠钾氯协同转运蛋白-1(Na-K-2Cl cotransporter 1,NKCC1)抑制剂XH-6003,每日2次,共3 d。3 d后出舱检测。主要结局指标为脑含水量、血脑屏障通透性、脑组织形态学变化、水通道蛋白4(aquaporin-4,AQP4)和NKCC1表达量;次要结局指标为行为学、凋亡和氧化应激指标。结果成功建立HACE大鼠模型。与对照组相比,模型组脑含水量增加(P<0.0001),血脑屏障破坏(P<0.0001),出现学习记忆下降(P<0.001)和焦虑抑郁情绪(P<0.01);qPCR结果显示模型组大鼠脑组织NKCC1和AQP4表达明显增加(P<0.01),病理结果显示模型组大鼠海马出现神经元和神经胶质细胞损伤(P<0.01)。予以NKCC1抑制剂XH-6003后,脑含水量、BBB损伤、神经元和胶质细胞损伤均有逆转(P<0.05),大鼠脑组织NKCC1和AQP4表达下降(P<0.01),凋亡相关蛋白被抑制,氧化应激指标中仅还原型谷胱甘肽(glutathione,GSH)有改善(P<0.001)。XH-6003治疗后的大鼠仅在新物体探索时间上有一定的功能改善,其他行为学结果均呈现阴性。结论HACE伴有NKCC1/AQP4通路的激活,抑制该通路可减轻脑水肿和BBB破坏,改善神经元和神经胶质细胞损伤。XH-6003在细胞分子层面具有治疗脑水肿的潜力,但其对HACE相关行为障碍的改善效果尚需进一步探究。Objective To investigate the pathogenesis of high-altitude cerebral edema(HACE)and develop new therapeutic strategies.Methods Male Sprague-Dawley(SD)rats of 6 weeks old were selected and placed in a hypobaric chamber.The rats were exposed to the high-altitude environment of 7000 m above sea level for 3 days for HACE modeling.Whether the HACE model was successfully established in the rats was evaluated by measuring brain water content,the degree of disruption to the blood-brain barrier(BBB),and brain tissue Nissl staining.The experimental animals were divided into four groups,with 28 rats in each group.The blank control group was exposed to a normobaric and normoxic environment simulating the conditions at 500 m above sea level for 3 d.The other groups,including a model group(the HACE group),a bumetanide group(the positive control group),and a XH-6003 treatment group,were placed at an altitude of 7000 m above sea level and were injected with normal saline,bumetanide,and XH-6003,a new type of Na-K-2Cl cotransporter 1(NKCC1)inhibitor,via the tail vein,respectively,twice daily for 3 d.The experimental animals were taken out of the hypobaric chamber for testing after 3 d.The primary outcome measures included brain water content,BBB permeability,changes in brain tissue morphology,and the expression levels of aquaporin-4(AQP4)and NKCC1.The secondary outcome measures included behavioral changes,apoptosis,and oxidative stress markers.Results The HACE rat model was successfully established.The model group exhibited increased brain water content(P<0.0001),BBB disruption(P<0.0001),impairment in learning skills and memory(P<0.001),and anxiety/depression-like behaviors(P<0.01).qPCR results showed significantly increased expression of NKCC1 and AQP4 in the brain tissue of the model group(P<0.01).Pathology examination revealed neuronal and glial cell damage in the hippocampus of the model group(P<0.01).Treatment with XH-6003,the NKCC1 inhibitor,reversed brain water content,BBB disruption,and neuronal and glial cell damage to a c

关 键 词：高原脑水肿 神经损伤 XH-6003 NKCC1 

分 类 号：R594.3[医药卫生—内科学]