依达拉奉抑制线粒体分裂减轻大鼠脑缺血再灌注损伤

Edaravone alleviates cerebral ischemia-reperfusion injury in rats by inhibiting mitochondrial division

目的探讨依达拉奉抑制线粒体分裂减轻大鼠脑缺血再灌注损伤。方法成年雄性SD大鼠90只,3月龄,体重250~300 g,将大鼠随机分为假手术组(S组)、脑缺血再灌注组(I/R组)和依达拉奉预先给药组(E组)。I/R组和E组分别于脑缺血前1 h腹腔注射等容量生理盐水和依达拉奉10 mg/kg。采用线栓法制备大鼠脑缺血再灌注损伤模型,缺血2 h,再灌注24 h。再灌注24 h时,采用Longa评分法行神经功能缺损评分(NDS),TUNEL法检测脑组织神经细胞凋亡情况,计算凋亡指数(AI),ELISA法检测丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,Western blot法检测动力相关蛋白1(Drp 1)及细胞色素C(Cyt C)蛋白的表达,RT-PCR法检测Drp 1及Cyt C mRNA的表达。结果与S组比较,I/R组与E组NDS、AI、MDA的含量明显增加,而SOD活性显著下降,Drp 1及Cyt C蛋白和Drp 1及Cyt C mRNA表达水平显著升高(P<0.05);与I/R组比较,E组NDS、AI、MDA的含量明显减少,而SOD活性显著升高,Drp 1及Cyt C蛋白和Drp 1及Cyt C mRNA表达水平显著下降(P<0.05)。结论依达拉奉通过减轻氧化应激,抑制线粒体分裂,减少细胞凋亡和坏死,改善脑缺血再灌注损伤。

Objective To investigate the inhibition of mitochondrial division by edaravone on cerebral ischemia-reperfusion injury in rats.Methods Ninety male SD rats,aged 3 months,weighing 250~300 g,were randomly divided into sham operation group(group S),ischemia-reperfusion group(group I/R)and edaravone treatment group(group E).Group E was given edaravone of 10 mg/kg by intraperitoneal injection 1 h before cerebral ischemia,Group I/R was given the equal volume of normal saline.The ischemia-reperfusion model was performed by thread occlusion of middle cerebral artery for 2 h,and then 24 h reperfusion.The Longa score was used to evaluate neurological deficit at 24 h of reperfusion,the TdT-mediated dUTP nick end labeling(TUNEL)was applied to observe the neuronal apoptosis,the apoptosis index(AI)was calculated.The enzyme linked immune sorbent assay(ELISA)was used to evaluate the level of superoxide dismutase(SOD)and malondialdehyde(MDA).Western blotting was used to analyze the protein expression of dynamin-related protein 1(Drp 1)and cytochrome C(Cyt C).Reverse transcription-polymerase chain reaction(RT-PCR)were used to detect the expression of Drp 1 and Cyt C.Results Compared with group S,the neurological deficit score,apoptosis index and MDA content of group I/R and group E were increased significantly,while SOD activity decreased significantly;Drp 1 and Cyt C protein,Drp 1 and Cyt C mRNA expression levels increased significantly(P<0.05);Compared with the group I/R,the neurological deficit score,apoptosis index and MDA content in group E were significantly decreased,while the activity of SOD was significantly increased;Drp 1 and Cyt C protein,Drp 1 and Cyt C mRNA expression levels decreased significantly(P<0.05).Conclusion Edaravone can alleviate oxidative stress,inhibit mitochondrion division,reduce apoptosis and necrosis,and improve cerebral ischemia-reperfusion injury.