Akt/GSK3β信号通路在黄芩苷改善机械通气致脑损伤小鼠认知功能中的作用  

Role of Akt/GSK3βsignal pathway in baicalin improving the cognitive function of mice with brain injury induced by mechanical ventilation

作　　者：瞿敏[1] 孙文波[1] 姚忠岩[1] 张秀青 张会武[2] 陈磊 齐子龙[1] Qu Min;Sun Wenbo;Yao Zhongyan;Zhang Xiuqing;Zhang Huiwu;Chen Lei;Qi Zilong(Department of Anesthesiology,Cangzhou Central Hospital,Cangzhou 061001,China;Pediatric Endoscopy Room of Cangzhou Central Hospital,Cangzhou 061001,China;Department of Pediatric Surgery,Cangzhou Central Hospital,Cangzhou 061001,China)

机构地区：[1]沧州市中心医院麻醉科,沧州061001 [2]沧州市中心医院小儿窥镜室,沧州061001 [3]沧州市中心医院儿外科,沧州061001

出　　处：《中华行为医学与脑科学杂志》2022年第12期1073-1079,共7页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：沧州市重点研发计划指导项目(204106029);河北省卫健委科研项目(20181528)。

摘　　要：目的探讨黄芩苷对机械通气致脑损伤小鼠认知功能的影响及其机制。方法C57BL6小鼠72只,体质量20~25 g,8~12周龄,采用随机数字表法分为对照组(C组)、机械通气组(V组)、黄芩苷组(B组)和黄芩苷+Akt抑制剂MK-2206组(BM组),每组18只。C组小鼠不行机械通气,自主呼吸空气6 h;V组小鼠给予机械通气6 h;B组与BM组小鼠机械通气前30 min腹腔注射黄芩苷100 mg/kg,BM组小鼠机械通气前60 min脑室内注射Akt抑制剂MK-2206300μg/kg。每组随机选取6只小鼠分别于机械通气前1 d、机械通气后3 d和7 d时采用Morris水迷宫实验检测小鼠的学习记忆能力。机械通气后1 d,每组处死6只小鼠,取脑组织,采用TUNEL法检测海马CA1区神经元凋亡情况,计算凋亡指数。于机械通气后1 d,每组处死6只小鼠,取海马,采用Western blot法检测caspase-3、caspase-9、Akt、p-Akt、GSK-3β和p-GSK-3β的蛋白表达。采用SPSS 22.0软件对数据行统计分析,多组间比较采用重复测量方差分析和单因素方差分析,进一步两两比较采用LSD-t检验。结果Morris水迷宫结果显示,四组小鼠时间×分组交互作用不显著(F=1.14,P>0.05),时间主效应和组间主效应均显著(F=47.36,59.65,均P<0.05)。机械通气后3 d、7 d时,V组小鼠逃避潜伏期高于C组(均P<0.05),穿越平台次数均低于C组(均P<0.05);机械通气后3 d、7 d时,B组小鼠逃避潜伏期均低于V组(P<0.05),穿越平台次数均高于V组(均P<0.05);BM组小鼠在机械通气后3 d,7 d逃避潜伏期均高于B组(均P<0.05),穿越原平台位置次数均低于B组(均P<0.05)。TUNEL和Western blot检测结果显示,四组小鼠海马神经元凋亡指数及凋亡相关蛋白caspase-3、caspase-9表达水平均差异有统计学意义(F=51.42,41.21,40.19,均P<0.05)。V组小鼠海马神经元凋亡指数[(40.6±3.9)%],海马caspase-3(4.93±0.92)、caspase-9(4.81±0.88)表达水平均高于C组[(13.7±1.4)%,1.87±0.27,1.71±0.25,均P<0.05],B组小鼠海马神经元凋亡指�Objective To investigate the effect of baicalin on cognitive function of mice with brain injury induced by mechanical ventilation and its mechanism.Methods Seventy two C57BL6 mice,weighing 20-25 g,aged 8-12 weeks,were randomly divided into control group(group C),mechanical ventilation group(group V),baicalin group(group B),baicalin+Akt inhibitor MK-2206 group(group BM)according to random number table method,with 18 in each group.Mice in group C did not have mechanical ventilation and breathed air independently for 6 hours.Mice in group V received mechanical ventilation for 6 hours.Mice in group B and group BM were intraperitoneally injected with baicalin 100 mg/kg 30 minutes before mechanical ventilation,and mice in group BM were injected intraventricular with Akt inhibitor MK-2206300μg/kg 60 minutes before mechanical ventilation.Six mice in each group were randomly selected to test their learning and memory abilities by Morris water maze test 1st day before mechanical ventilation and 3rd day and 7th day after mechanical ventilation.One day after mechanical ventilation,six mice in each group were killed,and the brain tissue was taken.TUNEL method was used to detect the neuronal apoptosis in hippocampal CA1 area,and the apoptosis index was calculated.One day after mechanical ventilation,six mice in each group were killed,and the hippocampus was taken,Western blot was used to detect the protein expressions of caspase-3,caspase-9,Akt,p-Akt,GSK-3βand p-GSK-3β.SPSS 22.0 software was used for statistical analysis of data,repeated measure ANOVA and one-way ANOVA were used for comparison between multiple groups.LSD-t test was used for further pairwise comparison.Results The results of water maze test showed that the time and group interaction of the four groups were not significant(F=1.14,P>0.05),the main effect of time and group were both significant(F=47.36,59.65,both P<0.05).At 3rd day and 7th day after mechanical ventilation,the escape latencies of mice in group V were higher than those in group C(both P<0.05),and

关 键 词：黄芩苷 机械通气 脑损伤 凋亡 Akt/GSK-3β信号通路 

分 类 号：R285.5[医药卫生—中药学]