新型硫化氢供体对癫痫大鼠模型海马Nrf2蛋白表达和SOD活性的影响  

Effects of a New Type of Hydrogen Sulfide Donor on the Expression of Nrf2 Protein and SOD Activity in the Hippocampus of Epileptic Rats Model

作　　者：张艺泷 黄宝怡 黄文幸 钟宇婷 詹芷晴 邓镇 汪鹏 刘国辉 朱晓琴[1,2] 苏涛 Zhang Yilong;Huang Baoyi;Huang Wenxing(Institute of Neuroscience and the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China;Department of Physiology,School of Basic Medical Sciences,Guangzhou Medical University Guangzhou 511436,China)

机构地区：[1]广州医科大学附属第二医院神经科学研究所,广州510260 [2]广州医科大学基础医学院生理学教研室,广州511436

出　　处：《华中科技大学学报（医学版）》2021年第6期700-704,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.82071444);广东省基础与应用基础基金资助项目(No.2020A1515010985);2021年广东省科技创新战略专项资金立项项目(No.pdjh2021b0419);2020年度广州医科大学大学生实验室开放项目;广州医科大学2020-2021大学生科技创新项目(No.2020A037)。

摘　　要：目的探究新型硫化氢(H_(2)S)供体对癫痫大鼠海马核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)蛋白表达和超氧化物歧化酶(superoxide dismutase,SOD)活性的影响。方法腹腔注射戊四氮(pentylenetetrazole,PTZ)制备急性大鼠癫痫模型,并经侧脑室给予不同的药物处理。将实验大鼠随机分为4组:对照组(Control组),致痫组(epilepsy,EP组),硫化氢干预组(H_(2)S+EP组),硫化氢干预+Nrf2蛋白抑制剂(鸦胆子苦醇,brusatol)组(H_(2)S+Brusatol+EP组)。行为学观察评估大鼠癫痫发作情况,BL-420系统监测各组大鼠脑电波的变化。分光光度法检测海马组织SOD活性变化,Western blot检测各组海马组织Nrf2蛋白的表达。结果行为学观察显示,与EP组相比,H_(2)S+EP组癫痫发作潜伏期延长,发作持续时间缩短,发作级别降低;与H_(2)S+EP组相比,H_(2)S+Brusatol+EP组大鼠癫痫发作潜伏期缩短,发作级别增加。脑电图结果显示,与EP组相比,H_(2)S+EP组癫痫发作波幅减小;与H_(2)S+EP组相比,H_(2)S+Brusatol+EP组大鼠癫痫发作波幅增大。分光光度法结果显示,与EP组相比,H_(2)S+EP组大鼠海马组织SOD活性显著增高(P<0.05);与H_(2)S+EP组相比,H_(2)S+Brusatol+EP组SOD活性显著降低(P<0.05)。Western blot结果显示,与EP组相比,H_(2)S+EP组海马组织Nrf2蛋白表达水平显著升高(P<0.05);与H_(2)S+EP组相比,H_(2)S+Brusatol+EP组Nrf2蛋白表达水平显著下降(P<0.05);与EP组相比,H_(2)S+Brusatol+EP组Nrf2蛋白表达无显著性差异(P>0.05)。结论H_(2)S可以抑制癫痫发作,作用机制可能与其上调Nrf2蛋白的表达,增高SOD活性,降低氧化应激水平有关。Objective To explore the effects of a novel hydrogen sulfide donor on the expression of nuclear factor erythroid 2-related factor 2(Nrf2)protein and the activity of superoxide dismutase(SOD)in the hippocampus of epileptic rats.Methods An acute rat epilepsy model was prepared by intraperitoneal injection of pentylenetetrazole(PTZ),and different drug treatments were given via the lateral ventricles.The experimental rats were randomly divided into four groups:control group,epilepsy group(EP group),hydrogen sulfide intervention group(H_(2)S+EP group),and hydrogen sulfide intervention+Nrf2 protein inhibitor(brusatol)group(H_(2)S+Brusatol+EP group).The epileptic seizures of rats were observed by behavioral studies,and the brain electrical changes of rats in each group were monitored by the BL-420 system.The changes of SOD activity in the hippocampus were detected spectrophotometrically,and the expression of Nrf2 protein in the hippocampus of each group was determined by Western blotting.Results Behavioral observations showed that compared with the EP group,the rats in the H_(2)S+EP group had a longer seizure latency,shorter seizure duration,and lower seizure level;compared with the H_(2)S+EP group,the rats in the H_(2)S+Brusatol+EP group had a shorter seizure latency and increased seizure level.The EEG results revealed that compared with the EP group,the H_(2)S+EP group showed a decrease in seizure wave amplitude and a reduction in the number of epileptiform waves;compared with the H_(2)S+EP group,the H_(2)S+Brusatol+EP group had a larger seizure wave amplitude and more epileptiform waves.Spectrophotometry showed that compared with the EP group,the SOD activity of the hippocampus in the H_(2)S+EP group was significantly increased(P<0.05);compared with the H_(2)S+EP group,the SOD activity of the H_(2)S+Brusatol+EP group was significantly decreased(P<0.05).The results of Western blotting showed that compared with the EP group,the expression of Nrf2 protein in the hippocampus of the H_(2)S+EP group was remarkably increase

关 键 词：硫化氢 癫痫 NRF2 超氧化物歧化酶 氧化应激 

分 类 号：R742.1[医药卫生—神经病学与精神病学]