AMPA受体对丙泊酚诱导新生大鼠海马线粒体损伤的调控作用  

Regulation of AMPA receptor on propofol induced hippocampal mitochondrial injury in neonatal rats

作　　者：王晨旭[1] 周琰[1] 谢克亮[1] 元元[1] 王国林[1] 于泳浩[1] Wang Chenxu;Zhou Yan;Xie Keliang;Yuan Yuan;Wang Guolin;Yu Yonghao(Department of Anesthesiology,Tianjin Medical University General Hospital,Tianjin Research Institute of Anesthesiology,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院麻醉科,天津市麻醉学研究所,天津300052

出　　处：《中华危重病急救医学》2023年第2期189-194,共6页Chinese Critical Care Medicine

基　　金：天津市科技计划项目(20JCQNJC01050);天津市教委科研计划项目(2019KJ201)。

摘　　要：目的探讨丙泊酚是否对新生大鼠海马线粒体造成损伤,以及兴奋性氨基酸受体AMPA受体的调控作用。方法将48只7日龄SD大鼠按随机数字表法分为对照组、丙泊酚组、丙泊酚+AMPA受体激动剂AMPA组(丙泊酚+AMPA组)和丙泊酚+AMPA受体抑制剂CNQX组(丙泊酚+CNQX组),每组12只。丙泊酚各组大鼠腹腔注射30 mg/kg丙泊酚,对照组注射生理盐水3 mg/kg;各组均于首次给药后每隔20 min给予首剂量的1/2,每日3次,连续注射3 d。丙泊酚+AMPA组和丙泊酚+CNQX组分别于首次给药后经左侧脑室注射1 g/L的AMPA或CNQX 5μL。各组给药3 d后处死大鼠取脑组织,采用蛋白质免疫印迹试验(Western blotting)检测海马AMPA受体谷氨酸受体(GluR1、GluR2)亚单位的总蛋白(T)及膜蛋白(M)表达量,并计算二者比值(M/T);采用Western blotting检测线粒体动力相关蛋白-1(DRP-1)、磷酸化DRP-1(p-DRP-1)和线粒体融合蛋白2(Mfn2)表达;同时测定海马三磷酸腺苷(ATP)含量及ATP相关酶活性。结果与对照组相比,丙泊酚处理后大鼠海马GluR1的表达及其M/T比值均明显升高,而GluR2的表达及其M/T比值则明显降低,ATP含量及ATP相关酶活性明显下降,同时DRP-1表达及其磷酸化水平明显升高,而Mfn2表达明显下降,说明反复多次腹腔注射30 mg/kg丙泊酚可导致神经细胞线粒体损伤。与丙泊酚组比较,加入AMPA激动剂后,GluR1表达及其M/T比值进一步升高〔T-GluR1蛋白(T-GluR1/β-actin):2.41±0.29比1.72±0.11,M-GluR1蛋白(M-GluR1/β-actin):1.18±0.15比0.79±0.09,M/T比值:0.78±0.12比0.46±0.08,均P<0.01〕;GluR2表达明显升高〔T-GluR2蛋白(T-GluR2/β-actin):0.65±0.13比0.30±0.14,P<0.01;M-GluR2蛋白(M-GluR2/β-actin):0.17±0.05比0.13±0.07,P>0.05〕,但其M/T比值进一步降低(0.27±0.10比0.41±0.08,P<0.05);ATP相关酶活性进一步降低,ATP含量进一步下降(μmol/g:0.32±0.07比0.70±0.10,P<0.01);线粒体DRP-1表达及其磷酸化水平进一步升高〔DRP-1蛋白(DRP-1/GAPDH):2.7Objective To investigate whether propofol can cause injury to hippocampal mitochondria in neonatal rats and the regulation of excitatory amino acid receptor AMPA receptor.Methods Forty-eight Sprague-Dawley(SD)rats aged 7 days were randomly divided into control group,propofol group,propofol+AMPA receptor agonist AMPA group(propofol+AMPA group)and propofol+AMPA receptor inhibitor CNQX group(propofol+CNQX group),with 12 rats in each group.The rats in the propofol groups were intraperitoneally injected with 30 mg/kg propofol,while in control group with 3 mg/kg normal saline.Each group was given 1/2 of the first dose every 20 minutes after the first administration,three times a day,for three consecutive days.The rats in the propofol+AMPA group and the propofol+CNQX group were injected with 1 g/L AMPA or CNQX 5μL through left ventricle after the first administration.Three days after administration,the rats were sacrificed to obtain brain tissue.Western blotting was used to determine the expression of AMPA receptor glutamate receptors(GluR1,GluR2)subunit totally(T)and on membrane(M)in hippocampus.The expression of dynamin-related protein-1(DRP-1)and phosphorylated-DRP-1(p-DRP-1)and mitofusin 2(Mfn2)related to mitochondrial fission and fusion were determined.The adenosine triphosphate(ATP)content and ATPase activity were determined.Results Compared with the control group,GluR1 expression and its M/T ratio were significantly increased after treatment of propofol,GluR2 expression and its M/T ratio were significantly decreased,the ATP content and ATP-related enzyme activity were decreased significantly,while the expression of DRP-1 and its phosphorylation was significantly increased,and the expression of Mfn2 was significantly decreased.The changes indicated that repeated intraperitoneal injection of 30 mg/kg propofol leading to the injury of mitochondria in neural cells.Compared with the propofol group,the GluR1 expression and its M/T ratio further increased after AMPA agonist administration[T-GluR1 protein(T-GluR1/β-act

关 键 词：丙泊酚 AMPA受体 线粒体 新生大鼠 

分 类 号：R614[医药卫生—麻醉学]