桃叶鸦葱黄酮调控神经元自噬和凋亡减轻大鼠脑缺血损伤  

The flavonoids of Scorzonera sinensis lipsch regulate neuronal autophagy and apoptosis to reduce cerebral ischemia injury in rats

作　　者：褚建波 王征 娄阳云 苏钠 唐洗敏 郭涛 CHU Jianbo;WANG Zheng;LOU Yangyun;SU Na;TANG Ximin;GUO Tao(College of Pharmacy,Hangzhou Medical College,Hangzhou 310053,China;School of Medicine,Kunming University of Science and Technology,Kunming 650500,China;School of Basic Medical Sciences,Dali University,Dali 671003,China)

机构地区：[1]杭州医学院药学院,杭州310053 [2]昆明理工大学医学院,昆明650500 [3]大理大学基础医学院,大理671003

出　　处：《生命的化学》2022年第5期1026-1034,共9页Chemistry of Life

基　　金：云南省省级大学生创新创业训练计划项目(2021106740080);昆明理工大学医学院课外学术科技创新基金项目(2022210019)。

摘　　要：桃叶鸦葱(Scorzonera sinensis lipsch,SSL)黄酮在中国已广泛用作脑卒中的辅助治疗,但其潜在的药理及生化机制尚未完全了解,本研究旨在探讨SSL是否通过调节缺血性脑卒中半影区神经元的自噬和凋亡从而具有神经保护作用。构建SD大鼠大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)脑缺血再灌注模型,将大鼠随机分为5组:假手术组(Sham)、模型组(MCAO)、腹腔给药治疗组(MCAO+SSL)、侧脑室注射自噬抑制剂3-甲基腺嘌呤(3-methyladenine)干预组(MCAO+3-MA)、给药加干预组(MCAO+SSL+3-MA),每组6只。缺血再灌注损伤7 d后,通过蛋白免疫印迹法和实时荧光定量PCR(qRT-PCR)分别检测自噬蛋白Beclin-1、LC3在各组中的表达,双重免疫荧光评估脑缺血半影区自噬标记物LC3和凋亡标记物TUNEL在神经元中的阳性细胞百分比,并检测神经功能缺损评分和梗死体积。结果显示,SSL治疗可显著提高MCAO神经元中的自噬表达并降低细胞凋亡水平,脑梗死体积和神经功能缺损明显减轻,但是SSL的这些作用可以被自噬抑制剂3-MA抵消(P<0.05)。上述结果表明,SSL可以赋予针对缺血性脑卒中的神经保护作用,这种生物学功能与神经元中的自噬增强和凋亡减少有关。Scorzonera sinensis lipsch(SSL)flavonoids have been widely used as an adjuvant treatment for stroke in China,but its potential pharmacological and biochemical mechanisms are not yet fully understood.This study aims to investigate whether SSL has a protective effect by regulating the autophagy of neurons in the penumbra of ischemic stroke.The sprague-dawley(SD)rat cerebral ischemia-reperfusion model was prepared by middle cerebral artery occlusion(MCAO),and the rats were randomly divided into 5 groups:sham operation group(Sham),model group(MCAO),abdominal cavity administration treatment group(MCAO+SSL),lateral ventricle autophagy inhibitor intervention group(MCAO+3-MA),administration plus intervention group(MCAO+SSL+3-MA),with 6 rats in each group.7 days after ischemia-reperfusion injury,the autophagy protein and autophagy genes of Beclin-1 and LC3 were detected by Western blot(WB)and quantitative real-time PCR(qRT-PCR),respectively.Double immunofluorescence to evaluate the percentage of positive cells in neurons of the autophagy marker LC3 and the apoptosis marker TUNEL in the penumbra of cerebral ischemia,and the neurological deficit score and infarct volume were detected.The results showed that SSL treatment could significantly increase the expression of autophagy and decrease the level of apoptosis in MCAO neurons,and significantly reduce cerebral infarction volume and neurological deficits,but these effects can be offset by the autophagy inhibitor 3-MA(P<0.05).The above results suggest that SSL can confer neuroprotective effects against ischemic stroke.This biological function is related to increased autophagy and decreased apoptosis in neurons.

关 键 词：脑缺血 桃叶鸦葱 黄酮 自噬 凋亡 半影区 神经保护 

分 类 号：R285.5[医药卫生—中药学]