高压氧对缺氧缺血性脑损伤新生大鼠脑神经元的保护作用及其对ERK1/2信号通路的影响  

Protective effects of hyperbaric oxygen on neurons in neonatal rats with hypoxic-ischemic brain damage and its impact on the extracellular signal-regulated protein kinase 1/2 signaling pathway

作　　者：郭晋伟[1] 夏莉[1] 栗红[1] 阴怀清[1] Guo Jinwei;Xia Li;Li Hong;Yin Huaiqing(Department of Pediatrics,The First Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学第一医院儿科,太原030001

出　　处：《中华航海医学与高气压医学杂志》2024年第6期733-738,共6页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：山西省卫生健康委科研计划项目(2022027)。

摘　　要：目的探究高压氧(HBO)处理对缺氧缺血性脑损伤(HIBD)新生大鼠脑神经元的保护作用及其对胞外信号相关蛋白激酶1/2(ERK 1/2)信号通路的影响。方法按随机数字表法将60只7 d龄健康SD大鼠分为假手术组、模型组、干预组、阻断剂组,每组15只。参考Rice-Vannucci法构建新生大鼠HIBD模型。假手术组大鼠暴露颈动脉后直接缝合;模型组大鼠造模后不给予任何治疗措施;干预组大鼠造模结束1 h后给予HBO干预;阻断剂组大鼠在每次HBO干预之前,尾静脉注射0.05 mg/kg ERK1/2阻断剂U0126。干预结束后每组随机处死10只大鼠取材,采用TTC染色测定脑组织梗死体积;HE染色观察脑组织病理形态改变;TUNEL染色观察并统计脑组织神经元细胞凋亡率;Western blotting实验测定脑组织ERK1/2、p-ERK1/2、Bcl-2、Bax蛋白的表达;采用逆转录-聚合酶链式反应(RT-PCR)测定脑组织ERK1/2 mRNA、Bcl-2 mRNA、Bax mRNA水平。4组剩余大鼠继续饲养14 d后,采用Morris水迷宫实验检测大鼠的学习记忆功能。结果与假手术组比较,其他3组大鼠脑组织梗死体积比及神经元细胞凋亡率明显增大(P<0.05);与模型组比较,干预组大鼠脑组织梗死体积比及神经元细胞凋亡率明显减小(P<0.05);与干预组比较,阻断剂组大鼠脑梗死体积比及神经元细胞凋亡率明显增大(P<0.05)。假手术组大鼠脑神经元细胞形态正常,整齐密集,组织结构完整;模型组大鼠脑神经元细胞数量减少,排列紊乱,形成空洞,可见胞质空泡,细胞核固缩;干预组大鼠可见脑组织神经元局部损伤,但程度较模型组轻;阻断剂组大鼠脑组织神经元损伤较干预组严重,与模型组接近。与假手术组比较,干预组大鼠脑组织p-ERK1/2蛋白表达水平明显升高(P<0.05),其他3组大鼠脑组织Bcl-2蛋白表达水平明显降低,Bax蛋白表达水平明显升高(P<0.05);与模型组比较,干预组大鼠脑组织p-ERK1/2和Bcl-2蛋白表达水平明显升高,Bax�Objective To explore the protective effects of hyperbaric oxygen(HBO)on neurons in neonatal rats with hypoxic-ischemic brain damage(HIBD)and its impact on the extracellular signal-regulated protein kinase 1/2(ERK1/2)signaling pathway.Methods Sixty 7-day-old healthy SD rats were divided in accordance with a random number table into a sham operation group,a model group,an intervention group,and a blocker group,with 15 rats in each group.A neonatal rat HIBD model was constructed with reference to the Rice-Vannucci model.In the sham operation group,the carotid arteries of rats were exposed and then directly sutured;in the model group,no treatment was given to the rats after modeling;in the intervention group,HBO intervention was given to the rats one hour after modeling;in the blocker group,before each HBO intervention,0.05 mg/kg ERK1/2 blocker U0126 was injected into the cauda vein of rats.After the intervention,10 rats in each group were randomly selected and then euthanized for sampling.The infarct volume of brain tissue was measured through TTC staining;the pathological morphological changes of brain tissue were observed by HE staining;the apoptosis rate of neuronal cells in brain tissue was observed and counted by TUNEL staining;the expressions of ERK1/2,p-ERK1/2,Bcl-2,and Bax proteins in brain tissue were determined by Western Blot assay;the levels of ERK1/2 mRNA,Bcl-2 mRNA,and Bax mRNA in brain tissue were determined by reverse transcription polymerase chain reaction(RT-PCR).The remaining rats in the 4 groups were fed for another 14 days,and the learning and memory abilities of the rats were detected by Morris water maze experiment.Results Compared with the sham operation group,the cerebral infarct volume ratio and neuronal cell apoptosis rates of the other three groups of rats were significantly increased(P<0.05);compared with the model group,the intervention group rats showed a significant decrease in the cerebral infarct volume ratio and neuronal cell apoptosis rates(P<0.05);compared with the intervention g

关 键 词：高压氧 缺氧缺血性脑损伤 胞外信号相关蛋白激酶1/2 神经元保护 凋亡 

分 类 号：R651.15[医药卫生—外科学]