食欲素-A对高氧诱导幼鼠脑损伤后认知功能的影响  

Effect of orexin-A on cognitive function of neonatal rats after hyperoxia-induced brain injury

作　　者：付志杰 张乐[1] 李艳娜[1] 魏雨婷 庞志路[3] Fu Zhijie;Zhang Le;Li Yanna;Wei Yuting;Pang Zhilu(Department of Anesthesiology,Pain and Perioperative Medicine,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450002,China;Department of Anesthesiology,Second Affiliated Hospital of Harbin Medical University,Harbin 150000,China;Department of Anesthesiology,Henan Provincial People′s Hospital,Zhengzhou 450002,China)

机构地区：[1]郑州大学第一附属医院麻醉与围术期医学部,450002 [2]哈尔滨医科大学第二附属医院麻醉科,150000 [3]河南省人民医院麻醉科,郑州450002

出　　处：《中华实验外科杂志》2021年第7期1291-1294,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关项目(LHGJ20190205)。

摘　　要：目的:探讨食欲素-A(OA)对高氧所致幼鼠脑损伤后认知功能的影响及其机制。方法:清洁级Sprague-Dawley幼鼠63只,体重40~50 g,将其随机数字表法分为3组:高氧组(HO组,n=21),连续7 d吸入高浓度氧(>80%)并腹腔注射磷酸盐缓冲液(PBS)0.5 ml;高氧并腹腔注射食欲素A组(OA组,n=21),连续7 d吸入高浓度氧(>80%)并腹腔注射OA 30μg/kg[以PBS稀释成0.5 ml];对照组(C组,n=21),连续7 d吸入空气并腹腔注射PBS 0.5 ml。模型建立成功后第1天进行Morris水迷宫试验,连续6 d后测定海马组织湿/干重(W/D)比值,Real-time PCR法测定海马白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)和冷诱导RNA结合蛋白(CIRP)mRNA表达的水平,蛋白质印迹法(Western blot)检测海马IL-1β、TNF-α和CIRP的蛋白含量。行为学表现指标组间比采用重复测量设计的方差分析,其他指标组间比采用单因素方差分析。结果:HO组和OA组幼鼠平台所在象限滞留时间[(17.2±4.4)、(25.9±3.6)s]、平均游泳速度[(11.69±0.83)、(13.92±0.98)cm/s]均低于C组,差异有统计学意义(F=74.600、283.000,P<0.05),逃逸潜伏期[(48.4±8.6)、(38.6±8.1)、(33.9±7.4)、(19.6±5.7)、(22.9±5.5),(41.3±6.4)、(33.4±5.5)、(27.5±4.7)、(15.5±4.5)、(16.4±4.3)s]、海马W/D(5.94±0.36、5.27±0.33)、海马CIRP(3.35±0.63、1.96±0.30)、TNF-α(2.89±0.45、1.34±0.37)和IL-1β(2.66±0.53、1.73±0.45)mRNA表达及其蛋白含量[(0.86±0.23)、(0.61±0.19),(0.82±0.26)、(0.57±0.22),(0.78±0.18)、(0.52±0.15)pg/ml]均高于C组,差异有统计学意义(F=14.200、15.620、14.480、5.273、30.280、57.840、72.590、66.660、32.210、32.710、23.520、47.010,P<0.05);OA组幼鼠平台所在象限滞留时间、平均游泳速度均高于HO组(F=74.600、283.000,P<0.05),逃逸潜伏期、海马W/D比值、海马IL-1β、TNF-α和CIRP mRNA表达及其蛋白含量均低于HO组(F=14.200、15.620、14.480、5.273、30.280、57.840、72.590、66.660、32.210、32.710、23.520、47.010,P<0.05)。结论:OObjective To investigate the effect of orexin-A(OA)on cognitive function of neonatal rats after brain injury caused by hyperoxia and related molecular mechanisms.Methods STotally,63 clean Sprague-Dawley rats with body weight of 40 to 50 g were randomly divided into three groups according to the random number table method:hyperoxic group(HO group,n=21),given high concentration of oxygen(>80%)and intraperitoneal injection of 0.5 ml phosphate buffer saline(PBS)for 7 consecutive days;OA group[hyperoxia+intraperitoneal injection of 30μg/kg OA(diluted with PBS to 0.5 ml)for 7 consecutive days,n=21];control group(group C,n=21)given air inhalation and intraperitoneal injection of 0.5 ml PBS for consecutive 7 days.Morris water maze test was performed on the first day after the successful establishment of the model.After 6 consecutive days,the wet/dry weight ratio(W/D)of the hippocampus was determined.The mRNA expression levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and cold-induced RNA binding protein(CIRP)in the hippocampus were detected by Real-time polymerase chain reaction(PCR).The protein expression of IL-1β,TNF-αand CIRP in the hippocampus was detected by Western blotting.The intergroup ratio of behavioral performance indicators was analyzed by repeated measurement design,and the intergroup ratio of other indicators was analyzed by one-way ANOVA.Results The residence time in the quadrant of the platform[(17.2±4.4),(25.9±3.6)s]and the average swimming speed[(11.69±0.83),(13.92±0.98)cm/s]in HO and OA groups were significantly lower than those in group C(F=74.600,283.000,P<0.05).In HO and OA groups,the escape latency[(48.4±8.6),(38.6±8.1),(33.9±7.4),(19.6±5.7),(22.9±5.5),(41.3±6.4),(33.4±5.5),(27.5±4.7),(15.5±4.5),(16.4±4.3)s],W/D[(5.94±0.36),(5.27±0.33)]of the hippocampus,the mRNA expression level of hippocampal CIRP(3.35±0.63,1.96±0.30),TNF-α(2.89±0.45,1.34±0.37)and IL-1β(2.66±0.53,1.73±0.45)mRNA and the expression of proteins[(0.86±0.23),(0.61±0.19),(0.82±0.26),(0.57

关 键 词：食欲素-A 炎症 凋亡 脑损伤 认知功能 

分 类 号：R651.15[医药卫生—外科学]