硒代胱氨酸减轻小鼠短暂局灶性脑缺血再灌注损伤的研究  

Effect of selenocysteine on transient focal cerebral ischemia/reperfusion injury in mice

作　　者：王琨 倪鸣岳 王琰 孙磊[1] 孔德焕[1] 倪庆宾[1] Wang Kun;Ni Mingyue;Wang Yan;Sun Lei;Kong Dehuan;M Qingbin(Postdoctoral Research Station of Taian Citv Central Hospital,Taian 271000,China;Department of Biochemistry and Molecular Biology,Anhui Medical University,Hefei 230022,China)

机构地区：[1]泰安市中心医院博士后科研工作站,271000 [2]安徽医科大学生物化学与分子生物学教研室,合肥230022

出　　处：《中华行为医学与脑科学杂志》2020年第10期880-885,共6页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：山东省自然科学基金项目(H0906);山东省博士后创新项目(202003063)。

摘　　要：目的:探讨硒代胱氨酸(SeC)对小鼠短暂局灶性脑缺血再灌注(transient focal cerebral ischemia/reperfusion,tFCI/R)损伤的神经保护作用及机制。方法:线栓阻塞昆明小鼠大脑中动脉1 h后拔除,制作tFCI/R模型。通过多普勒血流监测和神经功能缺损评分筛选手术合格小鼠40只,按照随机数字表法分为SeC治疗组和tFCI/R模型对照组;另选20只作为假手术组(Sham组,小鼠经历手术全程而不阻塞大脑中动脉)。SeC治疗组小鼠给予术后0 h、24 h、48 h各1次腹腔注射SeC溶液(2 mg/30 g);模型组和Sham组小鼠给予同样方式注射等量生理盐水。于术后24 h、48 h、72 h每组各随机抽取6只小鼠进行神经功能缺损评分及神经行为学测试;之后所有小鼠麻醉处死取脑,行氯代三苯基四氮唑(TTC)染色测脑梗死体积,湿-干重法测脑水肿程度变化,Tunel-DAPI染色观察神经元凋亡情况,Western blot检测活性caspase-3和PARP表达,并检测小鼠脑组织GSH、GSH-Px表达情况。结果:Sham组小鼠未见神经功能缺损表现,术后24 h、48 h、72 h的tFCI/R组Zea Longa评分[(3.67±0.52)分,(3.33±0.52)分,(2.17±0.41)分]均比Sham组[(0.50±0.55)分,(0.67±0.52)分,(0.33±0.52)分]显著提高(t=10.26,8.86,6.81,均P<0.05),而SeC治疗组评分[(2.50±0.55)分,(1.67±0.82)分,(0.83±0.75)分]则比tFCI/R组显著降低(t=3.74,4.19,3.84,均P<0.05)。行为学评测结果与Zea Longa评分一致。术后72 h TTC染色显示:与tFCI/R组[(24.69±2.25)%]比较,SeC治疗组[(11.89±1.64)%]脑梗死体积明显缩小,差异有统计学意义(t=10.28,P<0.05);Sham组无梗死灶形成。湿-干重测量显示:与Sham组相比,tFCI/R组左侧脑组织含水量[(85.87±1.36)%]明显增加(t=8.73,P<0.05),SeC治疗组左侧脑组织含水量[(81.06±1.07)%]与tFCI/R组比较下降,差异有统计学意义(t=6.22,P<0.05)。Tunel-DAPI染色、Western blot结果表明,SeC处理显著下调了caspase-3和PARP的裂解,从而抑制了神经元凋亡。脑组织GSH含量检测证实,Objective:To investigate the neuroprotective effect and mechanism of selenocysteine(SeC)on transient focal cerebral ischemia/reperfusion(tFCI/R)injury in mice.Methods:A tFCI/R model was established in Kunming mice after 1 hour of middle cerebral artery occlusion.Forty mice qualified for surgery were screened by doppler blood flow monitoring and neurological function defect score,and were randomly assigned to the SeC treatment group and the tFCI/R control group,while another 20 mice were selected as the Sham operation control group(underwent the whole operation without middle cerebral artery obstruction).Mice in SeC treatment group were given SeC solution intraperitoneal injection(2 mg/30 g)once at 0 h,24 h and 48 h after surgery,and mice in the tFCI/R group and Sham group were injected with the same amount of saline in the same manner.At 24 h,48 h and 72 h after surgery,6 mice were randomly selected from each group for neurological impairment score and neurobehavioral test.Then the volume of cerebral infarction was measured by TTC staining,the degree of cerebral edema was measured by wet-dry weight method,the apoptosis of neurons was observed by Tunel-DAPI staining,the activity of caspase-3 and PARP were detected by Western blot,and the expressions of GSH and GSH-PX in mouse brain tissues were detected.Results:The Zea Longa scores of tFCI/R group((3.67±0.52),(3.33±0.52),(2.17±0.41)points)at 24h,48h and 72h after surgery were significantly higher than that of Sham group((0.50±0.55),(0.67±0.52),(0.33±0.52))(t=10.26,8.86,6.81,all P<0.05).The scores of SeC treatment group((2.50±0.55),(1.67±0.82),(0.83±0.75))were significantly lower than that of tFCI/R group(t=3.79,4.19,3.84,all P<0.05).The behavioral assessment results were consistent with the Zea Longa score.TTC experiment 72 h after surgery showed that no infarction lesion was formed in Sham group.Compared with tFCI/R group((24.69±2.25)%),SeC treatment group((11.89±1.64)%)had significantly reduced cerebral infarction volume,and the difference was statist

关 键 词：硒代胱氨酸 短暂局灶性脑缺血再灌注 氧化应激 细胞凋亡 

分 类 号：R965[医药卫生—药理学]