PKA-CREB信号通路在七氟烷减轻CPB诱发大鼠认知功能障碍中的作用  

Role of PKA-CREB signaling pathway in sevoflurane-induced reduction of cognitive impairment caused by cardiopulmonary bypass in rats

作　　者：石磊[1] 李亚南[2] 刘祥[1] 陈岩[1] 张琦 Shi Lei;Li Yanan;Liu Xiang;Chen Yan;Zhang Qi(Department of Anesthesiology,Hebei Children′s Hospital,Shijiazhuang 050031,China;Department of Anesthesiology,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China)

机构地区：[1]河北省儿童医院麻醉科,石家庄050031 [2]河北医科大学第三医院麻醉科,石家庄050051

出　　处：《中华麻醉学杂志》2022年第2期166-170,共5页Chinese Journal of Anesthesiology

基　　金：2022年度河北省医学科学研究课题(20220712,20221174)。

摘　　要：目的:评价蛋白激酶A(PKA)-环磷酸腺苷反应元件结合蛋白(CREB)信号通路在七氟烷减轻CPB诱发大鼠认知功能障碍其中的作用。方法:健康雄性SD大鼠40只,4月龄,体重300~350 g,采用随机数字表法分为4组(n=10):对照组(C组)、CPB组、CPB+七氟烷组(CS组)和CPB+七氟烷+PKA抑制剂H89组(CSH组)。CSH组侧脑室注射H895μl后,CS组和CSH组大鼠暴露于2.4%七氟烷中1 h,然后CPB组、CS组和CSH组建立无血预充心脏不停跳CPB模型60 min。于CPB结束后2 d时采用旷场试验评估大鼠自主运动能力。于CPB结束后3 d时采用Morris水迷宫实验检测大鼠认知功能。水迷宫结束后断头取脑,分离海马组织,采用流式细胞术检测神经元凋亡率,Western blot法检测PKA、磷酸化CREB(p-CREB)和脑源性神经营养因子(BDNF)的表达。结果:4组旷场试验运动速度、路程及中心停留时间比较差异无统计学意义(P>0.05)。与C组比较,其余3组逃避潜伏期延长,穿越原平台次数减少,原平台象限停留时间缩短,海马神经元凋亡率升高,PKA、p-CREB和BDNF表达下调(P<0.05);与CPB组比较,CS组逃避潜伏期缩短,穿越原平台次数增加,原平台象限停留时间延长,海马神经元凋亡率降低,PKA、p-CREB和BDNF表达上调(P<0.05),CSH组上述指标差异无统计学意义(P>0.05);与CS组比较,CSH组逃避潜伏期延长,穿越原平台次数减少,原平台象限停留时间缩短,海马神经元凋亡率升高,PKA、p-CREB和BDNF表达下调(P<0.05)。结论:七氟烷可通过激活PKA-CREB信号通路,减轻海马神经元凋亡,从而减轻CPB诱发大鼠认知功能障碍。Objective To evaluate the role of protein kinase A(PKA)-cyclic adenosine monophosphate response element binding protein(CREB)signaling pathway in sevoflurane-induced reduction of cardiopulmonary bypass(CPB)-induced cognitive impairment in rats.Methods Forty healthy male Sprague-Dawley rats,aged 4 months,weighing 300-350 g,were divided into 4 groups(n=10 each)using a random number table method:control group(group C),group CPB,CPB+sevoflurane group(group CS)and CPB+sevoflurane+PKA inhibitor H89 group(group CSH).After H895μl was injected into the lateral ventricle in group CSH,the rats in group CS and group CSH were exposed to 2.4%sevoflurane for 1 h,and then the CPB model of beating heart without blood priming for 60 min was developed in CPB,CS and CSH groups.The autonomic movement ability was evaluated using the open field test at 2nd day after CPB.Morris water maze test was used to assess the cognitive function at 3rd day after CPB.The rats were sacrificed after the Morris water maze test,the brain was removed and the hippocampal tissues were isolated for determination of the apoptosis rate of hippocampal neurons(by flow cytometry)and expression of PKA,phosphorylated CREB(p-CREB)and brain-derived neurotrophic factor(BDNF)(by Western blot).Results There was no significant difference in movement speed,distance and time of staying at the central region among the four groups(P>0.05).Compared with group C,the escape latency was significantly prolonged,the number of crossing the original platform was reduced,the time of staying at the original platform quadrant was shortened,the apoptosis rate of hippocampal neurons was increased,and the expression of PKA,p-CREB and BDNF was down-regulated in the other three groups(P<0.05).Compared with group CPB,the escape latency was significantly shortened,the number of crossing the original platform was increased,the time of staying at the original platform quadrant was prolonged,the apoptosis rate of hippocampal neurons was decreased,and the expression of PKA,p-CREB and BDNF was

关 键 词：环AMP依赖性蛋白激酶类 CAMP反应元件结合蛋白质 麻醉药 吸入 心肺转流术 认知障碍 

分 类 号：R614[医药卫生—麻醉学]