摘要
目的探讨水通道蛋白-9(AQP-9)在缺氧缺血性脑病(HIE)新生小鼠认知功能障碍中的作用及其机制。方法将90只昆明新生小鼠随机分为对照组、模型组和AQP-9组,每组30只。对照组小鼠正常喂养,不建立HIE模型,模型组和AQP-9组小鼠采用电凝法闭合颈总动脉建立HIE模型;建模成功后30 min,AQP-9组小鼠给予单次腹腔注射20μg AQP-9,对照组和模型组小鼠腹腔注射等量的生理盐水。分别于干预后第1、3、7天,每组选取10只小鼠颈椎脱臼处死,取海马组织。采用反转录聚合酶链反应法检测对照组和模型组小鼠海马组织中AQP-9 mRNA表达,免疫荧光法检测小鼠海马组织中AQP-9蛋白(对照组和模型组)和Ki-67蛋白(对照组、模型组和AQP-9组)表达,干预后第3~7天处死小鼠前进行Morris水迷宫实验检测3组小鼠空间学习记忆能力。结果干预后第1、3、7天对照组小鼠海马组织中AQP-9 mRNA和蛋白的相对表达量比较差异无统计学意义(P>0.05)。模型组小鼠干预后第3天时海马组织中AQP-9 mRNA和蛋白的相对表达量显著高于干预后第1、7天(P<0.05),模型组小鼠干预后第1天与第7天时海马组织中AQP-9 mRNA和蛋白的相对表达量比较差异无统计学意义(P>0.05);干预后第1、3、7天,模型组小鼠海马组织中AQP-9 mRNA和蛋白的相对表达量显著高于对照组(P<0.05)。对照组小鼠干预后第1、3、7天海马组织中Ki-67蛋白相对表达量比较差异无统计学意义(P>0.05);模型组和AQP-9组小鼠干预后第7天时海马组织中Ki-67蛋白相对表达量显著高于干预后第1、3天(P<0.05),模型组和AQP-9组小鼠干预后第1、3天时海马组织中Ki-67蛋白相对表达量比较差异无统计学意义(P>0.05);干预后第1、3、7天,模型组小鼠海马组织中Ki-67蛋白相对表达量显著高于对照组(P<0.05),AQP-9组小鼠海马组织中Ki-67蛋白相对表达量显著高于对照组和模型组(P<0.05)。干预后第3~7天,对照组和AQP-9组小鼠Morris水迷宫实验逃避潜伏期均逐渐缩短(P<0.05),而模型组小鼠Morris水迷宫实验逃避潜伏期无显著变化(P>0.05);干预后第3、4、5天,3组小鼠Morris水迷宫实验逃避潜伏期比较差异无统计学意义(P>0.05);干预后第6、7天时,模型组小鼠Morris水迷宫实验逃避潜伏期显著长于对照组(P<0.05);干预后第6天,AQP-9组与对照组小鼠Morris水迷宫实验逃避潜伏期比较差异无统计学意义(P>0.05);干预后第7天,AQP-9组小鼠Morris水迷宫实验逃避潜伏期显著短于模型组(P<0.05)。结论HIE小鼠海马组织中AQP-9表达上调,外源性AQP-9蛋白可以改善HIE小鼠的空间学习记忆能力,其机制可能与外源性AQP-9蛋白促进Ki-67表达有关。
Objective To investigate the role and mechanism of aquaporin-9(AQP-9)in cognitive dysfunction of neonatal mice with hypoxic-ischemic encephalopathy(HIE).Methods Ninety Kunming neonatal mice were randomly divided into control group,model group and AQP-9 group,with 30 mice in each group.The mice in the control group were fed normally,and the HIE model was not established.The mice in the model group and the AQP-9 group were used to establish HIE model by closing common carotid artery with electrocoagulation.Thirty minutes after the successful modeling,the mice in the AQP-9 group were given a single intraperitoneal injection with 20μg AQP-9 for intervention,and the mice in the control group and the model group were intraperitoneally injected with the same amount of normal saline.On the first,third and seventh day after the intervention,10 mice in each group were sacrificed by cervical dislocation,and the hippocampus were collected.The expression of AQP-9 mRNA in hippocampus of mice in the control group and model group was detected by reverse transcription polymerase chain reaction,and the expressions of AQP-9 protein(control group and model group)and Ki-67 protein(control group,model group and AQP-9 group)in hippocampus of mice were detected by immunofluorescence method.Before killing the mice,the spatial learning and memory ability of mice in the three groups was evaluated by the Morris water maze test from the third to seventh day after the intervention.Results There was no significant difference in the relative expressions of AQP-9 mRNA and protein in hippocampus of mice in the control group on the first,third and seventh day after the intervention(P>0.05).The relative expressions of AQP-9 mRNA and protein in hippocampus of mice in the model group on the third day was significantly higher than those on the first and seventh day after the intervention(P<0.05).There was no significant difference in the relative expressions of AQP-9 mRNA and protein in hippocampus of mice in the model group on the first day and the seventh day after intervention(P>0.05).On the first,third and seventh day after the intervention,the relative expressions of AQP-9 mRNA and protein in hippocampus of mice in the model group were significantly higher than those in the control group(P<0.05).There was no significant difference in the relative expression of Ki-67 protein in hippocampus of mice in the control group on the first,third and seventh day after intervention(P>0.05).The relative expression of Ki-67 protein in hippocampus of mice in the model group and AQP-9 group on the seventh day after intervention was significantly higher than that on the first and third day after intervention(P<0.05).There was no significant difference in the relative expression of Ki-67 protein in hippocampus of mice in the model group and AQP-9 group on the first and third day after intervention(P>0.05).On the first,third and seventh day after the intervention,the relative expression of Ki-67 protein in hippocampus of mice in the model group was significantly higher than that in the control group(P<0.05),and the relative expression of Ki-67 protein in hippocampus of mice in the AQP-9 group was significantly higher than that in the control group and model group(P<0.05).From the third day to the seventh day after the intervention,the escape latency of Morris water maze test of mice in the control group and AQP-9 group decreased gradually(P<0.05),but there was no significant change in the escape latency of Morris water maze test of mice in the model group(P>0.05).On the third,fourth and fifth day after the intervention,there was no significant difference in the escape latency of Morris water maze test of mice among the three groups(P>0.05).On the sixth and seventh day after the intervention,the escape latency of Morris water maze test of mice in the model group was significantly longer than that in the control group(P<0.05).There was no significant difference in the escape latency of Morris water maze test of mice between the AQP-9 group and the control group on the sixth day after the intervention(P>0.05).The escape latency of Morris water maze test of mice in the AQP-9 group was significantly shorter than that in the model group on the seventh day after the intervention(P<0.05).Conclusion The expression of AQP-9 is up-regulated in the hippocampus of HIE mice.Exogenous AQP-9 protein can improve the spatial learning and memory ability of HIE mice,and the mechanism may be related to exogenous AQP-9 protein promoting the expression of Ki-67.
作者
巴庆华
BA Qinghua(Department of Neurology,People's Hospital of Henan University of Chinese Medicine,People's Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China)
出处
《新乡医学院学报》
CAS
2022年第7期606-611,共6页
Journal of Xinxiang Medical University
