摘要
目的评价瞬时受体电位香草酸4(TRPV4)在右美托咪定改善机械通气致脑损伤小鼠认知功能降低中的作用。方法清洁级健康雄性C57BL6小鼠90只,体质量20~25 g,8~12周龄,采用随机数字表法分为5组(n=18):对照组(C组)、机械通气组(V组)、HC-067047组(H组)、右美托咪定组(D组)和右美托咪定+GSK1016790A组(DG组)。C组不行机械通气,自主呼吸空气6 h;V组机械通气6 h;H组机械通气3和6 h时分别脑室内注射TRPV4阻断剂HC-06704710 mmol;D组和DG组机械通气前30 min腹腔注射右美托咪定50μg/kg;DG组机械通气前60 min脑室内注射TRPV4激动剂GSK1016790A 5μmol。每组随机取6只小鼠分别于机械通气前1 d、机械通气后3和7 d时行Morris水迷宫实验。机械通气后1 d,每组随机处死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。每组随机处死6只小鼠取海马组织,采用Western blot法检测TRPV4、丝氨酸-苏氨酸蛋白激酶(Akt)、p-Akt、Bcl-2、Bax和caspase-3的表达。结果与C组比较,V组和DG组机械通气后3和7 d逃避潜伏期延长,穿越原平台位置次数减少,海马TRPV4和caspase-3表达上调,p-Akt表达下调,Bcl-2/Bax比值降低,神经元凋亡指数升高(P<0.05);与V组比较,D组和H组机械通气后3和7 d逃避潜伏期缩短,穿越原平台位置次数增加,海马TRPV4和caspase-3表达下调,p-Akt表达上调,Bcl-2/Bax比值升高,神经元凋亡指数降低(P<0.05);与D组比较,DG组机械通气后3和7 d逃避潜伏期延长,穿越原平台位置次数减少,海马TRPV4和caspase-3表达上调,p-Akt表达下调,Bcl-2/Bax比值降低,神经元凋亡指数升高(P<0.05)。结论TRPV4参与了右美托咪定改善机械通气致脑损伤小鼠认知功能降低的过程,与其上调p-Akt表达,抑制海马神经元凋亡有关。
Objective To evaluate the role of transient receptor potential vanillic acid 4(TRPV4)in dexmedetomidine-induced improvement in cognitive function in mice with mechanical ventilator-caused brain injury.Methods Ninety clean-grade healthy male C57BL6 mice,weighing 20-25 g,aged 8-12 weeks,were divided into 5 groups(n=18 each)using a random number table method:control group(group C),mechanical ventilation group(group V),HC-067047 group(group H),dexmedetomidine group(group D),and dexmedetomidine+GSK1016790A group(group DG).In group C,the animals breathed air spontaneously for 6 h without mechanical ventilation.In group V,the animals were mechanically ventilated for 6 h.In group H,TRPV4 blocker HC-06704710 mmol was injected into the cerebral ventricle at 3 and 6 h of mechanical ventilation.In D and DG groups,dexmedetomidine 50μg/kg was intraperitoneally injected at 30 min before mechanical ventilation.In group DG,TRPV4 agonist GSK1016790A 5μmol was injected into the cerebral ventricle at 60 min before mechanical ventilation.Morris water maze test was performed on 6 mice in each group at 1 day before mechanical ventilation and 3 and 7 days after mechanical ventilation.Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation,and the brain tissue was taken for determination of the neuronal apoptosis in hippocampal CA1 area by TUNEL method,and the apoptosis index was calculated.Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation,and the hippocampal tissues were taken for determination of the expression of TRPV4,serine-threonine protein kinase(Akt),phosphorylated Akt(p-Akt),Bcl-2,Bax and caspase-3 by Western blot.Results Compared with group C,the escape latency was significantly prolonged and the number of crossing the original platform was reduced at 3 and 7 days after mechanical ventilation,the expression of TRPV4 and caspase-3 was up-regulated,the ratio of Bcl-2/Bax was decreased,and the apoptosis index of neurons was increased in g
作者
瞿敏
孙文波
姚忠岩
张秀青
张会武
陈磊
齐子龙
黄冬冬
黄平
Qu Min;Sun Wenbo;Yao Zhongyan;Zhang Xiuqing;Zhang Huiwu;Chen Lei;Qi Zilong;Huang Dongdong;Huang Ping(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;Department of Medical Diagnosis,Cangzhou Medical College,Cangzhou 061001,China;Department of Gynecology,Cangzhou Central Hospital,Cangzhou 061001,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2023年第2期186-191,共6页
Chinese Journal of Anesthesiology
关键词
TRPV阳离子通道
右美托咪啶
呼吸
人工
脑损伤
认知
TRPV cation channels
Dexmedetomidine
Respiration,artificial
Brain injuries
Cognition
