Sun Jing, Li Yun-Zi, Ding Yin-Hui, Wang Jin, Geng Ji, Yang Huan, Ren Jie, Tang Jin-Yan, Gao Jing
Neurobiology Laboratory, School of Pharmacy, Jiangsu University, No. 301 Xuefu Road, Zhenjiang 212013, P R China.
Neurobiology Laboratory, School of Pharmacy, Jiangsu University, No. 301 Xuefu Road, Zhenjiang 212013, P R China.
Brain Res. 2014 Nov 17;1589:126-39. doi: 10.1016/j.brainres.2014.09.039. Epub 2014 Sep 22.
Oxidative stress and mitochondrial dysfunction are frequently implicated in the pathology of secondary neuronal damage following cerebral ischemia/reperfusion. Recent evidence suggests that gallic acid (GA) reverses oxidative stress in rat model of streptozotocin-induced dementia, but the roles and mechanisms of GA on cerebral ischemia/reperfusion injury remain unknown. Here we investigated the potential roles and mechanisms of GA in hypoxia/reoxygenation induced by sodium hydrosulfite (Na2S2O4) in vitro and cerebral ischemia/reperfusion induced by middle cerebral artery occlusion (MCAO) in vivo. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, 5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethylbenzimidazol carbocyanine iodide (JC-1), Dichlorofluorescin diacetate (DCF-DA) and MitoSOX fluorescent assay, Clark-type oxygen electrode, firefly luciferase assay, and calcium-induced mitochondrial swelling were conducted to detect cell death, mitochondrial membrane potential (MMP), intracellular and mitochondrial reactive oxygen species (ROS), oxygen consumption, ATP level, and mitochondrial permeability transition pore (MPTP) viability. We firstly find that modulation of the mitochondrial dysfunction is an important mechanism by GA attenuating hypoxia/reoxygenation insult. To further assess the effects of GA on cerebral ischemia/reperfusion injury, 2, 3, 5-triphenyl-tetrazolium chloride (TTC) staining, dUTP nick-end labeling (TUNEL) assay, and Cytochrome C (Cyt C) release were performed in MCAO rats. The results support that GA is useful against cerebral ischemia/reperfusion injury as a potential protective agent.
氧化应激和线粒体功能障碍常与脑缺血/再灌注后继发性神经元损伤的病理过程有关。最近的证据表明，没食子酸（GA）可逆转链脲佐菌素诱导的痴呆大鼠模型中的氧化应激，但GA在脑缺血/再灌注损伤中的作用和机制仍不清楚。在此，我们研究了GA在体外连二亚硫酸钠（Na2S2O4）诱导的缺氧/复氧及体内大脑中动脉闭塞（MCAO）诱导的脑缺血/再灌注中的潜在作用和机制。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法、5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑碳氰碘化物（JC-1）、二氯荧光素二乙酸酯（DCF-DA）和MitoSOX荧光法、Clark型氧电极、萤火虫荧光素酶法以及钙诱导的线粒体肿胀实验来检测细胞死亡、线粒体膜电位（MMP）、细胞内和线粒体活性氧（ROS）、氧消耗、ATP水平以及线粒体通透性转换孔（MPTP）活力。我们首先发现，调节线粒体功能障碍是GA减轻缺氧/复氧损伤的重要机制。为进一步评估GA对脑缺血/再灌注损伤的影响，对MCAO大鼠进行了2,3,5-三苯基氯化四氮唑（TTC）染色、脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）法检测及细胞色素C（Cyt C）释放检测。结果支持GA作为一种潜在的保护剂对脑缺血/再灌注损伤有效。
