Zhu Bei-Lei, Xie Chen-Long, Hu Ning-Ning, Zhu Xin-Bo, Liu Chun-Feng
Department of Neurology and Suzhou Clinical Research Center of Neurological Disease, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Department of Neurology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Behav Neurol. 2018 Aug 1;2018:5701719. doi: 10.1155/2018/5701719. eCollection 2018.
The aim of this study was to explore the role of DL-3-n-butylphthalide (NBP) in cerebral ischemia-reperfusion injury (CIRI) mice model. The involvement of extracellular signal-regulated kinase (ERK) signaling pathway was also investigated.
All mice were divided into five groups: sham-operated group, CIRI group, NBP pretreatment group, NBP treatment group, and NBP pretreatment + treatment group. The CIRI mice model was established by the use of the Pulsinelli four-vessel occlusion method. Pretreatment mice received NBP (90 mg/kg/d) three times a day within four days before reperfusion by gavage. Treatment mice received NBP (90 mg/kg/d) three times a day within five days after reperfusion by gavage. We detected the infarction area, the neurological severity, and the superoxide dismutase and malondialdehyde levels. Furthermore, we observed the expressions of GRASP65, phosphorylation of GRASP65 (pGRASP65), ERK, and phosphorylation of ERK (pERK) by the use of Western blotting.
The result showed that the ERK pathway was activated in response to CIRI. NBP decreases the expressions of pERK and pGRASP65 following CIRI. Additionally, NBP could decrease MDA and increase SOD level in brain tissues. Decreased infarct volume was also observed in the NBP group. Thereby, NBP inhibited the activation of the ERK pathway induced by CIRI and reduced the GRASP65 phosphorylation.
The current finding suggested that NBP protected the cerebrum from CIRI mediated by inhibiting the ERK signaling pathway and subsequently reducing GRASP65 phosphorylation.
本研究旨在探讨丁苯酞（NBP）在脑缺血再灌注损伤（CIRI）小鼠模型中的作用。同时也研究了细胞外信号调节激酶（ERK）信号通路的参与情况。
将所有小鼠分为五组：假手术组、CIRI组、NBP预处理组、NBP治疗组和NBP预处理+治疗组。采用Pulsinelli四血管闭塞法建立CIRI小鼠模型。预处理小鼠在再灌注前4天内每天经口灌胃给予NBP（90mg/kg/d）3次。治疗小鼠在再灌注后5天内每天经口灌胃给予NBP（90mg/kg/d）3次。我们检测了梗死面积、神经功能严重程度以及超氧化物歧化酶和丙二醛水平。此外，我们通过蛋白质印迹法观察了GRASP65、GRASP65磷酸化（pGRASP65）、ERK和ERK磷酸化（pERK）的表达。
结果显示，ERK通路在CIRI反应中被激活。NBP可降低CIRI后pERK和pGRASP65的表达。此外，NBP可降低脑组织中丙二醛水平并提高超氧化物歧化酶水平。NBP组梗死体积也减小。因此，NBP抑制了CIRI诱导的ERK通路激活并降低了GRASP65磷酸化。
目前的研究结果表明，NBP通过抑制ERK信号通路并随后降低GRASP65磷酸化来保护大脑免受CIRI损伤。
