乙酰谷酰胺对脑外伤小鼠脑水肿程度及Omi/HtrA2信号通路的影响  

The effect of acetylglutamin on brain edema and mitochondrial Omi/HtrA2 signaling pathway in mice with traumatic brain injury

作　　者：罗明 黄英[1] 王旭辉[1] 杨超 Luo Ming;Huang Ying;Wang Xuhui(Chongming Branch,Xinhua Hospital,School of Medicine,Shanghai Jiao Tong University,Shanghai 202150)

机构地区：[1]上海交通大学医学院附属新华医院崇明分院,202150

出　　处：《卒中与神经疾病》2021年第6期605-609,617,共6页Stroke and Nervous Diseases

基　　金：上海市卫生和计划生育委员会科研计划项目(编号为2018Y0016)。

摘　　要：目的探讨乙酰谷酰胺对脑外伤小鼠脑水肿程度及Omi/HtrA2信号通路的影响。方法应用C57BL/6小鼠进行实验,随机分为治疗组、模型组和假手术组,每组10只小鼠;模型组和治疗组小鼠麻醉后选择中线旁开2 mm处和右侧冠状缝后1 mm作为撞击点,将其头皮切开,钻1个5 mm直径的骨孔,应用Feeney自由落体损伤装置,找25 cm处应用40 g重击锤自由落体撞击,打击深度为5 mm,将鼠皮缝合;假手术组小鼠仅将其头皮切开,并不进行自由落体击打;造模30 min后治疗组小鼠腹腔注射乙酰谷酰胺,其余2组大鼠注射1 mL生理盐水,连续治疗3 d;第3 d小鼠深度麻醉后断头处死,并将小鼠的海马组织和大脑组织进行分离;对受损侧大脑组织含水量进行检测;对各组小鼠的海马组织的细胞凋亡情况进行检测;应用免疫印迹(Western blot,WB)对小鼠海马组织中的聚腺苷二磷酸核糖聚合酶(Polyadenosine diphosphate ribose polymerase,PARP)、前体半胱氨酸天冬氨酸蛋白酶9(Precursor caspase 9,Pro-caspase-9),Pro-caspase-3,X染色体连锁的调亡抑制蛋白(X-linked apoptosis suppressor protein,XIAP)以及Omi/TtrA2蛋白水平进行检测;应用四肽荧光底物法对海马组织中Caspase-9和Caspase-3蛋白活性进行检测。结果小鼠颅脑损伤后损伤侧脑组织的含水量明显升高;与模型组的(82.34±0.28%)比较,治疗组的脑组织含水量(78.28±0.78%)有所降低(P<0.05);假手术组的脑组织含水量为(77.37±0.49%),与治疗组的脑组织含水量相当(P>0.05);模型组的海马神经元凋亡率明显高于假手术组(P<0.05),治疗组小鼠的海马神经元凋亡率要低于模型组(P<0.05);模型组海马组织PARP,Pro-caspase-9,Pro-caspase-3以及Omi/TtrA2水平较假手术组高(P<0.05),模型组海马组织的XIAP蛋白水平相对于假手术组有所降低(P<0.05);治疗组小鼠海马组织PARP,Pro-caspase-9,Pro-caspase-3以及Omi/TtrA2水平相对于模型组有所降低,XIAP蛋白水平有所升高(P<0.0Objective To study the effect of Acetylglutamine on brain edemaand Omi/HtrA2 signaling pathway in mice with traumatic brain injury.Methods C57 BL/6 mice wererandomly divided into treatment group,model group and sham,and 10 micefor each group.Theimpact pointwhere 2 mm side of the midline and 1 mm behind the right coronal suture wasdetermined.On the mice of model group and treatment group,their scalp was incised and abone holewith 5 mm diameter was drilledafter anesthesia.The Feeney free fall injury device was used to make the traumatic brain injury model.A hammer weighted 40 g fell verticallyfrom the heightof 25 cm,which made a 5 mm deepinjury,followed by suturing the scalp.The mice in the sham group only had their scalp incised,without injury on brain.The mice in the treatment group were injected intraperitoneally with acetylglutamine at 30 minutes after injury,while the mice in other two groups were injected with 1 mL 0.9%salineonce a day for 3 days.On the third day,the mice were decapitated after anesthesia,and the hippocampus and brain tissues were separated.The water content of the injured brain was tested;the apoptotic neuronsweredetected in the hippocampus.The proteins of polyadenosine diphosphate ribose polymerase(PARP),Precursor caspase(Pro-caspase)-9,Pro-caspase-3,X-linked apoptosis suppressor protein(XIAP)and Omi/TtrA2 were detected by western bolt,the activity of proteins Caspase-9 and Caspase-3 in hippocampus was detected by tetrapeptide fluorescent substrate method.Results After the brain injury in mice,the water content of the injured brain(82.34±0.28%)increased significantly in the model group,compared with thesham group(77.37±0.49%).But compared with model mice,it is decreased in the treatment group(78.28±0.78%)(P<0.05).There wasno significant difference in water content of the injured brain between the treatment group and the sham group(P>0.05).The percentage of apoptotic neuronsin hippocampus of model mice weresignificantly morethan that in sham mice(P<0.05),while itsin treatment group was si

关 键 词：乙酰谷酰胺 创伤性脑损伤 脑水肿 Omi/HtrA2信号通路 细胞凋亡 

分 类 号：R651.15[医药卫生—外科学]