五味子醇甲诱导缺血半影区神经元自噬提高脑卒中后神经保护  

Schisandrin A Improves Neuroprotection by Induction of Neuronal Autophagy in Ischemic Penumbra after Stroke

作　　者：臧瑞 杨继苹 郭涛 褚鑫 张雪玉 武煜明 邓仪昊 ZANG Rui;YANG Ji-Ping;GUO Tao;CHU Xin;ZHANG Xue-Yu;WU Yu-Ming;DENG Yi-Hao(Department of Human Anatomy and Histology Embryology,Basic Medical School,Yunnan University of Chinese Medicine,Kunming 650500,China;Department of Human Anatomy,Medicine School,Kunming University of Science and Technology,Kunming 650500,China)

机构地区：[1]云南中医药大学基础医学院人体解剖与组织胚胎学教研室,昆明650500 [2]昆明理工大学医学院解剖学教研室,昆明650500

出　　处：《中国生物化学与分子生物学报》2020年第5期544-551,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.81660383,No.81860411,No.81960418);云南省科技计划项目(No.2017FF117(-018),No.2017FF116(-022))资助。

摘　　要：五味子醇甲(Schisandra A,Sch A)是五味子中具有生物活性的木脂素化合物,其神经保护作用已在多种神经系统疾病动物模型中得到验证。然而,五味子醇甲是否能通过影响大鼠脑缺血半影区神经元自噬活性对脑缺血再灌注大鼠模型产生神经保护作用,尚缺乏系统研究。为探究Sch A对大鼠脑卒中后神经损伤及缺血半影区神经元自噬活性的影响,本研究将90只SD雄性大鼠随机分为假手术(Sham)组、模型组(MCAO)、Sch A低剂量组(40μg/kg)、Sch A中剂量组(80μg/kg)、Sch A高剂量组(160μg/kg),每组18只。线栓法制备大鼠大脑中动脉梗塞(middle cerebral artery occlusion,MCAO)模型,脑缺血持续90 min后进行再灌注,立即侧脑室给药,1/d,连续给药7 d。各组分别取6只大鼠进行神经功能评分后,取脑进行TTC染色检测脑梗死体积。另有6只大鼠取缺血半影区脑组织,通过Western印迹检测自噬相关蛋白质Beclin1、LC3-Ⅱ的表达水平。剩余6只大鼠脑组织用于免疫荧光双标,对Sch A改变的自噬活性进行细胞表达定位。研究结果显示,MCAO组大鼠脑梗死体积及神经功能损伤评分均显著高于Sham组(P<0.05),且Beclin1及LC3-Ⅱ的表达显著增加(P<0.05)。各Sch A治疗组大鼠脑梗死体积较未给药组显著减少(P<0.05),神经功能损伤得到明显改善(P<0.05)。同时,Sch A给药组Beclin1及LC3蛋白质表达水平明显升高(P<0.05),且免疫荧光双标显示该自噬活性改变主要呈现于神经元。以上结果表明,Sch A可显著减轻大鼠脑缺血再灌注损伤,该神经保护作用与其提高缺血半影区神经元自噬活性密切相关。Schisandra A(Sch A)is a bioactive lignan compound in Schisandra,whose neuroprotective effect has been demonstrated in animal models of various neurological diseases.However,there is still no systematic study showing whether Sch A produces neuroprotective effects on the cerebral ischemia reperfusion rat model by affecting neuronal autophagy activity in the ischemic penumbra of the rat brain.Therefore we investigated the efficacy of schisandra A(Sch A)on neural injury and autophagic activity of neurons in ischemic penumbra of rats with cerebral stroke.90 Male Sprague-Dawley rates were randomly divided into five groups:the Sham group,the middle cerebral artery occlusion(MCAO)model group,the Sch A low-dose group(40μg/kg),the Sch A medium-dose group(80μg/kg)and the Sch A high-dose group(160μg/kg).The rat model(MCAO)was prepared according to the modified ZeaLonga method.The cerebral ischemia was persistent for 90 min and then prepared for reperfusion.The Sch A was administrated by lateral ventricle immediately after onset of reperfusion once daily for seven days.Six rats in each group were randomly selected to perform neurological score,the infarction volume was then measured by TTC staining assays.The brain tissues in the penumbra from six rats in each group were gained to detect the expressions of autophagy-related protein of Beclin1 and LC3-Ⅱby western blotting in ischemic penumbra.The brain tissues from the rest six rats were collected to detect the LC3 expression and its cellular localization by immunofluorescence,and the result was represented by percentage of LC3-NeuN-positive cells.The results showed that the infarct volume and neurological score in the MCAO group were significantly higher than those in the sham group.Meanwhile,the expression levels of LC3-Ⅱand Beclin1 were markedly promoted.The infarct volume in each Sch A administration group was dramatically attenuated,compared with that in the non-administration group.Furthermore,the neurofunctional deficit was significantly alleviated,with the expre

关 键 词：五味子醇甲 脑缺血再灌注 自噬 神经保护 

分 类 号：R392[医药卫生—免疫学]