N-乙酰血清素对缺氧缺血性脑病乳鼠的神经保护作用及对水通道蛋白-4表达的影响  

Neuroprotective effect of N-acetylserotonin on hypoxic-ischemic encephalopathy and its influence on aquaporin-4in neonatal rats

作　　者：罗兴 方成志[1] 曾红兰 张丙宏[1] LUO Xing;FANG Chengzhi;ZENG Honglan;ZHANG Binghong(Department of Neonatology,Renmin Hospital of Wuhan University,Wuhan 430000,China)

机构地区：[1]武汉大学人民医院新生儿科,武汉430000

出　　处：《中华实用诊断与治疗杂志》2020年第11期1081-1085,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：中央引导地方科技发展专项资金[鄂财教发(2016)109];湖北省自然科学基金(WJ2019M184)。

摘　　要：目的制备缺氧缺血性脑病(hypoxic-ischemic encephalopathy,HIE)乳鼠模型,探讨N-乙酰血清素(N-acetylserotonin,NAS)对HIE乳鼠的神经保护作用及水通道蛋白-4(aquaporin-4,AQP-4)表达的影响。方法SD乳鼠60只随机分为假手术组、HIE组和NAS组各20只。HIE组、NAS组结扎左侧颈总动脉后用混合气体(体积分数8%O2和体积分数92%N2)缺氧2h制备HIE模型;假手术组仅分离左侧颈总动脉,既不结扎也不进行缺氧处理。造模后12、24、48h,NAS组腹腔注射体积分数5%NAS溶液10mg/kg,假手术组和HIE组分别注射等体积PBS溶液。造模后48h处死大鼠,取脑组织观察脑水肿情况并检测脑含水量,TTC染色检测脑梗死面积率,免疫荧光染色观察海马CA1区神经元特异核蛋白(neuron-specific nuclear protein,NeuN)、脑皮质APQ-4表达,Western blot法检测AQP-4蛋白相对表达量。结果假手术组未见脑水肿和脑梗死灶,海马CA1区神经元排列整齐、分布均匀,皮层AQP-4低表达;HIE组出现明显脑水肿和脑梗死灶,海马CA1区神经元稀疏、排列紊乱,皮层AQP-4高表达并与肿胀的星形胶质细胞共定位。HIE组、NAS组脑组织含水量[(88.40±2.38)%、(85.30±0.81)%]、AQP-4蛋白相对表达量(0.33±0.03、0.17±0.03)高于假手术组[(80.38±1.72)%、0.10±0.02],脑梗死面积率[(0.61±0.03)%、(0.36±0.05)%]大于假手术组(0),海马CA1区NeuN阳性神经元数目[(220.4±49.1)、(568.0±132.8)个/mm2]少于假手术组[(1160.0±224.1)个/mm2](P<0.05);NAS组脑梗死面积率小于HIE组,脑组织含水量、AQP-4蛋白相对表达量低于HIE组,海马CA1区NeuN阳性神经元数目多于HIE组(P<0.05)。结论NAS可通过抑制AQP-4的表达而减轻HIE乳鼠模型神经损伤。Objective To prepare neonatal rat models with hypoxic-ischemic encephalopathy(HIE)and to investigate the neuroprotective effect of N-acetylserotonin(NAS)on HIE neonatal rats and its influence on the expression of aquaporin-4(AQP-4).Methods Sixty SD neonatal rats were randomly divided into sham-operation group,HIE group and NAS group,with 20rats in each group.In HIE group and NAS group,the left common carotid artery was ligated and hypoxia was performed with mixed gas(8%O2and 92%N2)for 2hto prepare HIE models,while in sham-operation group,only the left common carotid artery was isolated without ligation or hypoxia treatment.NAS group was injected intraperitoneally with 10mg/kg 5%NAS in 12,24and 48hafter modeling,and sham-operation group and HIE group were injected with the same volume of PBS.The rats were sacrificed 48hafter modeling,and the brain tissue was taken to observe the brain edema and detect the brain water content.The percentage of cerebral infarction area was detected by TTC staining,and the expressions of neuron-specific nuclear protein(NeuN)in the hippocampal CA1region and AQP-4 in the cerebral cortex were detected by immunofluorescence staining.Western blot was used to detect the relative expression of AQP-4.Results In sham-operation group,no brain edema or cerebral infarction was found,the neurons in the hippocampal CA1region were neatly arranged and evenly distributed,and AQP-4was lowly expressed in the cortex.In HIE group,obvious brain edema and cerebral infarction were found,the neurons in the hippocampal CA1region were sparse and disordered,and AQP-4was overexpressed in the cortex and co-located with swollen astrocytes.The water contents in the brain tissue((88.40±2.38)%,(85.30±0.81)%)and the relative expressions of AQP-4protein(0.33±0.03,0.17±0.03)in HIE and NAS groups were higher than those in sham-operation group((80.38±1.72)%,0.10±0.02),the percentage of cerebral infarction area was larger in HIE group((0.61±0.03)%)and NAS group((0.36±0.05)%)than that in sham-operation group(0),and

关 键 词：缺氧缺血性脑病 脑水肿 N-乙酰血清素 水通道蛋白-4 乳鼠 

分 类 号：R742[医药卫生—神经病学与精神病学]