RhoA/ROCK2信号通路在多次七氟烷麻醉致新生大鼠远期认知功能障碍中的作用  

Role of RhoA/ROCK2 signaling pathway in multiple exposures to sevoflurane-induced long-term cognitive impairment in neonatal rats

作　　者：张琦 李亚南[2] 于家旭 尹春平[2] 赵娟[3] 侯志勇[4] 王秋筠[2] Zhang Qi;Li Yanan;Yu Jiaxu;Yin Chunping;Zhao Juan;Hou Zhiyong;Wang Qiujun(Department of Anesthesiology,Hebei Children′s Hospital,Shijiazhuang 050030,China;Department of Anesthesiology,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;Experimental Centre for Teaching of Hebei Medical University,Shijiazhuang 050011,China;Trauma Emergency Center,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050030 [2]河北医科大学第三医院麻醉科,石家庄050051 [3]河北医科大学教学实验中心,石家庄050011 [4]河北医科大学第三医院创伤急救中心,石家庄050051

出　　处：《中华麻醉学杂志》2022年第3期284-288,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(81771134);河北省自然科学基金面上项目(H2018206305)。

摘　　要：目的评价Rho亚家族蛋白A(RhoA)/Rho相关的卷曲连接蛋白激酶2(ROCK2)信号通路在多次七氟烷麻醉致新生大鼠远期认知功能障碍中的作用。方法SPF级健康新生SD大鼠60只,雌雄不拘,6日龄,体重12~20 g,采用随机数字表法分为3组(n=20):对照组(C组)、多次七氟烷麻醉组(S组)和RhoA/ROCK2信号通路抑制剂Y-27632组(Y组)。S组和Y组于出生后6、7和8 d时吸入3%七氟烷麻醉2 h;Y组于吸入七氟烷麻醉前腹腔注射Y-276325 mg/kg。于出生后35 d时行旷场实验评估自发活动能力;于出生后36 d时行Morris水迷宫实验评估认知功能。水迷宫实验结束后处死大鼠分离海马组织,采用流式细胞术检测神经元凋亡率和胞浆钙离子浓度([Ca^(2+)]_(i)),采用Western blot法测定磷酸化RhoA(p-RhoA)、ROCK2和裂解的caspase-3(cleaved-caspase-3)的表达水平,透射电镜下观察神经元超微结构。结果3组旷场实验运动速度、路程及旷场中心停留时间比较差异无统计学意义(P>0.05)。与C组比较,S组逃避潜伏期延长,穿越原平台次数减少,海马神经元凋亡率和[Ca^(2+)]_(i)升高,p-RhoA、ROCK2和cleaved-caspase-3表达上调(P<0.05),海马神经元发生病理学损伤。与S组比较,Y组逃避潜伏期缩短,穿越原平台次数增加,海马神经元凋亡率和[Ca^(2+)]_(i)降低,p-RhoA、ROCK2和cleaved-caspase-3表达下调(P<0.05),海马神经元病理学损伤减轻。结论多次七氟烷麻醉诱发新生大鼠远期认知功能障碍的机制与激活RhoA/ROCK2信号通路,诱发海马神经元凋亡有关。Objective To evaluate the role of RhoA/ROCK2 signaling pathway in multiple exposures to sevoflurane-induced long-term cognitive impairment in neonatal rats.Methods Sixty SPF healthy neonatal Sprague-Dawley rats of either sex,aged 6 days,weighing 12-20 g,were divided into 3 groups(n=20 each)using a random number table method:control group(group C),multiple exposures to sevoflurane group(group S)and RhoA/ROCK2 signaling pathway inhibitor Y-27632 group(group Y).Group S and group Y inhaled 3%sevoflurane for 2 h at days 6,7 and 8 after birth.In group Y,Y-276325 mg/kg was intraperitoneally injected before sevoflurane anesthesia.The spontaneous activity was evaluated by open field test on day 35 after birth.The cognitive function was detected by Morris water maze test at day 36 after birth.The rats were sacrificed after Morris water maze test,and the hippocampal tissues were isolated for determination of the apoptosis rate of hippocampal neurons and cytoplasmic calcium concentration([Ca^(2+)]_(i))(by flow cytometry)and expression of phosphorylated RhoA(p-RhoA),ROCK2 and cleaved-caspase-3(by Western blot)and for microscopic examination of the ultrastructure of hippocampal neurons(with a transmission electron microscope).Results There was no significant difference in movement speed,distance and time of stay in the open field center in the open field test among the three groups(P>0.05).Compared with group C,the escape latency was significantly prolonged,the number of crossing the original platform was reduced,the apoptosis rate of hippocampal neurons and[Ca^(2+)]_(i) were increased,the expression of p-RhoA,ROCK2 and cleaved-caspase-3 was up-regulated(P<0.05),and the pathological injury to hippocampal neurons was found in group S.Compared with group S,the escape latency was significantly shortened,the number of crossing the original platform was increased,the apoptosis rate of hippocampal neurons and[Ca^(2+)]_(i) were decreased,the expression of p-RhoA,ROCK2 and cleaved-caspase-3 was down-regulated(P<0.05),and the pathologi

关 键 词：RHO因子 RHO相关激酶类 麻醉药 吸入 婴儿 新生 认知功能障碍 

分 类 号：R614[医药卫生—麻醉学]