丹酚酸A通过Nrf2/HO-1途径减轻大鼠脑缺血再灌注损伤  

Salvianolic acid A alleviate the brain damage in rats after cerebral ischemia-reperfusion through Nrf2/HO-1 pathway

作　　者：张雯 宋俊科 闫蓉 何国荣 张雪 周启蒙 肖智勇[2] 周文霞[2] 杜冠华 

机构地区：[1]中国医学科学院、北京协和医学院药物研究所,北京市药物靶点研究与新药筛选重点实验室,北京100050 [2]军事医学科学院毒物药物研究所,抗毒药物与毒理学国家重点实验室,北京100850

出　　处：《药学学报》2016年第11期1717-1723,共7页Acta Pharmaceutica Sinica

基　　金：“重大新药创制”国家科技重大专项资助项目(2013ZX09508104,2013ZX09402203);抗毒药物与毒理学国家重点实验室开放课题资助项目(TMC201510);国家自然科学青年基金资助项目(81603100);北京协和医学院研究生创新基金资助项目(2015-1007-07)

摘　　要：研究丹酚酸A(SAA)减轻大鼠脑缺血再灌注损伤的机制。SD大鼠随机分组,线栓法制备大鼠大脑中动脉闭塞/再灌注(MCAO/R)模型,缺血1.5 h,再灌注24 h,对神经行为学缺损程度评分,TTC法测定大鼠脑梗死体积,Western blot测定脑组织胞核、胞浆Nrf2和全细胞HO-1蛋白含量。PC12细胞系制备缺糖缺氧复糖复氧(OGD/R)模型,缺糖缺氧6 h,复糖复氧24 h,MTT法检测细胞存活率,免疫荧光法测定Nrf2和HO-1表达,应用Nrf2 si RNA对SAA的作用机制进行研究。MCAO/R导致脑组织出现病理性损伤,OGD/R导致PC12细胞存活率显著降低。SAA(10和20 mg·kg^(-1))可使脑组织神经细胞损伤明显减轻,并且SAA(0.5和5μmol·L^(-1))能增加PC12细胞存活率。同时SAA能够促进脑组织及PC12胞核和胞浆中Nrf2蛋白表达,核转位率升高,HO-1蛋白表达增强。SAA具有抗脑缺血再灌注损伤的作用,其机制可能与激活Nrf2/HO-1信号途径,促进Nrf2合成和核转位,从而促进下游抗氧化蛋白HO-1的表达有关。The aim of present study is to investigate the protective effects and mechanism of salvianolic acid A （SAA） on cerebral ischemia-reperfusion injury in rats. The model was established with middle cerebral artery occlusion and reperfusion （MCAO/R） with ischemia for 1.5 h and reperfusion for 24 h in adult male SD rats. After the behavior assessment, TTC assay was used to calculate the infarct volume of rat brain; the distribution of Nrf2 in nuclear and cytoplasm and expression of HO-1 were detected by Western blot. The PC12 cells injury model was established with oxygen-glucose deprivation for 6 h and reintroduction for 24 h. Cell viability was determined with MTT assay, and the expression of Nrf2 and HO-1 were detected through immunofluorescence staining. The mechanisms were investigated in PC12 cells with Nrf2 knocking down by siRNA. SAA （10 and 20 mg·kg-1） significantly reduced the neuronal damage in MCAO/R model, and SAA （0.5 and 5 μmol·L^-1） increased cell viability in PC12 cells injury model. Meanwhile, the nuclear translocation of Nrf-2 and the expression of HO-1 were increased in PC12 cell and rats brain. SAA exhibited anti-cerebral ischemia-reperfusion effects. The mechanism may be related to activation of Nrf2/HO-1 signaling pathway, which promotes the synthesis and nuclear translocation of Nrf2 to enhance the expression of the antioxidant protein HO-1.

关 键 词：丹酚酸A 脑缺血再灌注 转录因子E2相关因子2 血红素氧合酶1 

分 类 号：R963[医药卫生—微生物与生化药学]