摘要
目的评估抑制泛素羧基末端水解酶L1(UCHL1)对小鼠脑缺血/再灌注损伤的影响。方法将雄性BALB/c小鼠随机分为假手术(sham)组、缺血/再灌注(I/R)组、UCHL1小干扰RNA(siRNA)组和scramble si RNA(control)组,每组10只小鼠。采用小鼠大脑中动脉闭塞(MCAO)60 min后再灌注24 h建立I/R模型。其中si RNA组和control组在MCAO前24 h将10μl UCHL1 si RNA或scramble si RNA通过侧脑室注入脑内。采用RTPCR和Western blotting方法检测各组小鼠脑组织中UCHL1表达;采用2,3,5-氯化三苯基四氮唑(TTC)染色法评估各组小鼠脑梗死体积和水肿率;采用神经行为学评分法评估各组小鼠神经症状评分。结果与sham组相比,I/R组缺血半影区UCHL1 m RNA和蛋白水平显著增高(P<0.05);而si RNA组UCHL1蛋白和mRNA表达明显降低(P<0.05);与此同时,I/R组小鼠脑梗死体积、水肿率及神经行为学损伤明显增加,而si RNA组小鼠脑梗死体积和水肿率及神经行为学损伤进一步加重(P<0.05)。结论抑制UCHL1可加重小鼠脑缺血/再灌注损伤。提示,MCAO后诱导UCHL1表达对小鼠脑缺血/再灌注损伤具有保护作用。
Objective To evaluate the effect of inhibition of ubiquitin carboxy terminal hydrolase L1(UCHL1) on cerebral ischemia/reperfusion injury in mice.Methods Male BALB/c mice were randomly divided into sham group,ischemia/reperfusion(I/R)group,UCHL1 small interfering RNA(si RNA)group and scramble si RNA(control)group,10 mice in each group.I/R model was established by reperfusion 24 hours after middle cerebral artery occlusion(MCAO)60 minutes.In the si RNA group and control group,10μl UCHL1 siRNA or scramble si RNA was injected into the brain through the lateral ventricle 24 hours before MCAO.The expression of UCHL1 was detected by RT-PCR and Western blotting;the volume of cerebral infarction and the rate of edema were assessed by 2,3,5-triphenyl tetrazolium chloride(TTC)staining;and the score of neurological symptoms was assessed by neurobehavioral scoring.Results Compared with the sham group,the level of UCHL1 m RNA and protein in ischemic penumbra of I/R group were significantly higher(P<0.05),while the expression of UCHL1 protein and m RNA in si RNA group were significantly lower(P<0.05);at the same time,the volume of cerebral infarction,edema rate and neurobehavioral damage in I/R group increased significantly,while the volume and edema rate of cerebral infarction and neurobehavioral damage in si RNA group further increased(P<0.05).Conclusion Inhibition of UCHL1 can aggravate the cerebral ischemia/reperfusion injury in mice,suggesting that the induction of UCHL1 after MCAO has a protective effect on the cerebral ischemia/reperfusion injury in mice.
作者
彭志锋
李晨旭
马国英
杨靖辉
PENG Zhi-feng;LI Chen-xu;MA Guo-ying;YANG Jing-hui(Department of Physiology,Medical College of Shanxi Datong University,Shanxi Datong 037009,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2021年第4期543-547,共5页
Acta Anatomica Sinica
基金
山西省基础研究项目(2015021178)
山西省大同大学博士科研启动经费(2014-B-01)。
