丹参酚酸B通过Nrf2/HO-1对脑缺血/再灌注损伤的保护作用研究  

Protective Effects of Salvianolic Acid B in Treatment of Cerebral Ischemia/Reperfusion through Nrf2/HO-1 Pathways

作　　者：卫永鲲[1] 郭晓华[2] 王波平 王卫东[1] 靳小力 李健康[3] 马慧玲[2] WEI Yong-kun GUO Xiao-hua WANG Bo-ping WANG Wei-dong JIN Xiao-li LI Jian-kang MA Hui-ling(HanZhong 3201 hospital, Hanzhong, Shaanxi, 723099, China Hanzhong vocational and technical college, Hanzhong, Shaanxi, 723002, China Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Xi＇an, Shaanxi, 710032, China)

机构地区：[1]汉中3201医院,陕西汉中723099 [2]汉中职业技术学院,陕西汉中723002 [3]第四军医大学西京医院药剂科,陕西西安710032

出　　处：《现代生物医学进展》2016年第35期6817-6821,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81173514);西京医院助推计划基础研究探索项目(XJZT15M20)

摘　　要：目的:研究丹参酚酸B对脑缺血/再灌注(Cerebral ischemia/reperfusion,CI/R)损伤的保护作用及机制。方法:通过结扎颈总动脉缺血2 h再灌注48 h复制CI/R模型,将实验大鼠随机分为假手术组、模型组、丹参酚酸B组,每组10只,培养大脑皮层神经细胞,分别给予0,10,25,50 umol/L的丹参酚酸B。通过2,3,5-氯化三苯基四氮唑蓝(TTC)染法测定大鼠脑梗死面积,Western Blot检测大鼠Nrf2和HO-1蛋白表达水平以及细胞中Nrf2和HO-1蛋白表达水平。再通过细胞缺氧缺糖模型,检测不同浓度丹参酚酸B对于细胞死亡率及细胞内ROS水平以及转染Nrf2或HO-1 si RNA后细胞死亡率及细胞内ROS水平。结果:与模型组比较,丹参酚酸B组的大鼠脑梗死面积明显减小,脑组织中Nrf2和HO-1蛋白表达水平均明显增加(P<0.05)。大脑皮层细胞中,随着丹参酚酸B浓度增加,细胞HO-1蛋白及细胞核中Nrf2蛋白表达水平逐渐提高,而细胞质中Nrf2蛋白表达水平逐渐降低(P<0.05)。细胞缺糖缺氧条件下,与对照组相比,丹参酚酸B组均能够降低细胞的死亡率及细胞内ROS水平,敲除Nrf2或HO-1后,丹参酚酸B组的细胞死亡率与细胞内ROS水平均有明显减低(P<0.05)。结论:丹参酚酸B对大鼠CI/R具有保护作用,其作用机制可能通过Nrf2/HO-1减轻CI/R所造成的氧化应激损伤。Objective： To study the protective effects and the mechanismof Salvianolic acid B on the cerebral ischemia/reperfusion injury in rats. Methods： An in vivo model of cerebral ischemia 2 h followed by reperfusion was made by reversibly ligating common carotid artery, The male SD rats were randomly divided into control group, vehicle ＋ UR group and Salvianolic acid B＋UR groupwith ten in each group. The primary of rat cortical neurons were cultured and give the dose of 0, 10, 25, 50 μmol/L Salvianolic acid B, respectively. Through the stained with solution of 2,3,5-triphenyltetrazolium chloride to calculate the infarct volume of brains, use western blot to ana- lyze the expression of protein of Nrt2 and HO-1 in rats and the expression of cytoplasmic and nuclear protein of Nrt2 and total protein of HO-1 in neurons. According to the oxygen-glucose deprivation （OGD） model, detect the cell death and cellular ROS in different dose of Salvianolic acid B and the cell death and cellular ROS after Nrf2 or HO-1 siRNA transfection. Results： Compared with vehicle group, the infarct volumes was significantly decreased in Salvianolic acid B group, and the expression of protein of Nrf2 and HO-1 was increased in rat brains （P〈0.05）. In rat cortical neurons, the expression of protein HO-1 and nuclear protein of Nrt2 was increased gradually, the expression of cytoplasmic protein of Nrf2 was reduced gradually （P〈0.05）. In OGD conditions, compared with control group, the cell death and cellular ROS were decreased in Salvianolic acid B group, knockdown of Nrt2 or HO-1, the cell death and cellular ROS were also significantly decreased in Salvianolic acid B group（P〈0.05）. Conclusions： Salvianolic acid B has the neuroprotection against I/R injury, and the mechanism may be related to its antioxidant activities through Nrf2/HO-lpathways.

关 键 词：脑缺血/再灌注损伤 丹参酚酸B Nrf2/HO-1 氧化应激 

分 类 号：R285[医药卫生—中药学] R-33[医药卫生—中医学]