氢对大鼠局灶性脑缺血再灌注时线粒体膜电位的影响  

Effect of hydrogen on mitochondrial membrane potential during focal cerebral ischemia-reperfusionin rats

作　　者：张鑫磊[1] 陈盼 夏裕宁 李雪梅[3] 谭永星[1] Zhang Xinlei, Chen Pan, Xia Yuning, Li Xuemei, Tan Yongxing,(1Department of lntensive Care Unit, Affiliated Hospital of Guilin Medical University, Guilin 541001, China; 2Department of Nursing, Affiliated Hospital of Guilin Medical University, Guilin 541001, China ; 3Graduate School of Guilin Medical University, Guilin 541001, China)

机构地区：[1]桂林医学院附属医院重症医学科,541001 [2]桂林医学院研究生院,541001 [3]桂林医学院附属医院重症护理部,541001

出　　处：《中华麻醉学杂志》2018年第5期598-601,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81260206）;广西壮族自治区自然科学基金（2013GXNSFAA019155,2015GXNSFAA139130）;广西壮族自治区卫生厅课题（Z2016390）

摘　　要：目的探讨氢对大鼠局灶性脑缺血再灌注时线粒体膜电位的影响。方法健康成年雄性SPF级sD大鼠84只，体重220～240g，采用随机数字表法分为3组（n=28）：假手术组（Sham组）、脑缺血再灌注组（I／R组）和氢气组（H，组）。I／R组和H，组采用大脑中动脉闭塞法制备局灶性脑缺血再灌注损伤模型。也组于再灌注即刻和再灌注12h时分别腹腔注射氢气10ml／kg。于再灌注24h时进行神经功能缺损评分；然后处死大鼠，取皮质区脑组织，光镜下观察病理学结果，采用TTC染色法测定脑梗死体积．TUNEL法测定神经细胞凋亡指数（AI）。JC-1法测定线粒体膜电位水平，Westernblot法测定Bcl-2及Bax的表达水平。结果与Sham组比较，I／R组和H，组神经功能缺损评分、脑梗死体积百分比和AI升高，线粒体膜电位水平降低，脑组织Bax表达上调，Bcl-2表达下调（P〈0．05）；与I／R组比较，H，组神经功能缺损评分、脑梗死体积百分比和AI降低，线粒体膜电位水平升高，脑组织Bax表达下调，Bcl-2表达上调（P〈0．05），脑组织病理学损伤减轻。结论氢减轻大鼠局灶性脑缺血再灌注损伤的机制可能与减少线粒体膜电位的耗散，抑制神经细胞凋亡有关。Objective To investigate the effect of hydrogen on mitoehondrial membrane potential during focal cerebral ischemiareperfusion （I/R） in rats. Methods Eightyfour healthy adult male SPF SpragneDawley rats, weighing 220240 g, were divided into 3 groups （n = 28 each） using a random num ber table： sham operation group （Sham group） , I/R group and hydrogen group （H2 group）. Focal cere bral I/R was produced by midcerebral artery occlusion in I/R and H2 groups. Hydrogen 10 ml/kg was intra peritoneally injected immediately after onset of reperfusion and at 12 h of reperfusion in group H2. Neurolog ical deficit was scored at 24 h of reperfusion. The rats were then sacrificed and brain tissues in cortex were removed for microscopic examination of the pathological changes and for determination of cerebral infarct size （by q^I＇C staining） , nerve cell apoptosis （by TUNEL） , mitochondrial membrane potential （by JC1 stai ning） and expression of Bcl2 and Bax （by Western blot）. Apoptotic index （AI） was calculated. Results Compared with Sham group, the neurological deficit score, percentage of cerebral infarct size and AI were significantly increased, the mitochondrial membrane potential was decreased, Bax expression in brain tissues was upregulated, and Bcl2 expression in brain tissues was downregulated in I/R and H2 groups（P〈0. 05）. Compared with I/R group, the neurological deficit score, percentage of cerebral infarct size and AI were significantly decreased, the mitochondrial membrane potential was increased, Bax expression in brain tissues was downregulated, and Bcl2 expression in brain tissues was upregulated （P〈0.05）, and the pathological changes of brain tissues were significantly attenuated in H2 group. Conclusion The mechanism by which hydrogen reduces focal cerebral I/R injury is related to decreasing dissipation of mito chondrial membrane potential and inhibiting nerve cell apoptosis in rats.

关 键 词：氢 再灌注损伤 脑 膜电位 线粒体 

分 类 号：R614[医药卫生—麻醉学]