人参皂苷Rg1对阿尔茨海默病大鼠模型脑片神经元自噬小体相关蛋白表达的影响  

Effects of ginsenoside Rg1 on the expression of neuronal autophagosome-related proteins in brain slices of a rat model of Alzheimer's disease

作　　者：贾健 张怡权[1,3] 乾坤 李玺 Jia Jian;Zhang Yi;Quan Qiankun;Li Xi(Department of Geriatrics,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710000,China;Department of Rehabilitation Medicine,Baoji Central Hospital,Baoji 721008,China;Department of Geriatrics,Xianyang Central Hospital,Xianyang 712000,China)

机构地区：[1]西安交通大学第二附属医院老年病科,710000 [2]宝鸡市中心医院康复医学科,721008 [3]咸阳市中心医院老年病科,712000

出　　处：《中华老年医学杂志》2022年第1期71-75,共5页Chinese Journal of Geriatrics

基　　金：陕西省自然科学基金(2017JQ8039)。

摘　　要：目的探讨人参皂苷Rg1对阿尔茨海默病(AD)大鼠模型脑片神经元自噬小体相关蛋白表达的影响及其分子机制。方法选取6周龄SD大鼠断头取脑制备海马脑片,将海马脑片随机分为空白对照组、模型组及低、中、高浓度Rg1组,每组10张脑片。模型组人工脑脊液中加入Aβ1-42(终浓度5μmol/L)处理2 h造模,低、中、高浓度Rg1组加入Aβ1-42(终浓度5μmol/L)作用2 h造模后分别加入Rg1致终浓度分别为60μmol/L、120μmol/L、240μmol/L作用3 h;空白对照组不给予任何干预药物。干预结束后采用苏木精-伊红(HE)染色观察各组海马脑片病理学改变情况;采用透射电镜检测各组海马脑片自噬小体情况;采用Western blot检测各组脑片自噬相关蛋白P62、LC3-Ⅱ/LC3-Ⅰ表达水平以及Aβ1-42和Shank蛋白表达水平。结果HE染色结果显示,模型组海马神经元排列紊乱,存在神经元死亡和缺失,Rg1各组海马神经元排列及缺失情况较模型组改善,以高浓度Rg1组改善最明显;透射电镜结果显示,模型组脑片自噬小体较空白对照组明显增多,Rg1各组脑片中自噬小体均较模型组减少。蛋白质印迹结果显示,与空白对照组比较,模型组脑片Shank1、P62和LC3-Ⅰ蛋白水平减少(均P<0.05),Aβ1-42、LC3-Ⅱ蛋白水平升高(均P<0.05);与模型组比较,各Rg1组脑片Shank1、P62和LC3-Ⅰ蛋白表达水平均升高(均P<0.05)、Aβ1-42、LC3-Ⅱ蛋白水平均降低(均P<0.05),其中以高浓度Rg1组最明显。结论人参皂苷Rg1可能通过上调AD大鼠模型海马脑片Shank1、P62、LC3-Ⅰ蛋白表达,抑制自噬作用,发挥大脑神经元的保护作用。Objective To explore the effects and molecular mechanisms of ginsenoside Rg1 on the expression of neuronal autophagosome-related proteins in a rat model of Alzheimer's disease(AD).Methods Six-week-old SD rats were decapitated to prepare hippocampal brain slices.The slices were randomly divided into the blank control group,the model group,the low-concentration,medium-concentration and high-concentration Rg1 groups,with 10 in each group.In the model group,Aβ1-42(final concentration:5μmol/L)was added into an artificial cerebrospinal fluid(CSF)for 2 h treatment.The low-concentration,medium-concentration and high-concentration Rg1 groups were treated with Aβ1-42(final concentration:5μmol/L)for 2 h,and then treated with Rg1(final concentrations:60μmol/L,120μmol/L,240μmol/L,respectively)for 3 h.The blank control group was not given any intervention drugs.At the end of intervention,histological changes of hippocampal brain slices in each group were examined via hematoxylin-eosin(HE)staining.Autophagosomes in hippocampal brain slices of each group were detected using transmission electron microscopy.The expression levels of autophagy-related proteins(P62,LC3-Ⅱ/LC3-Ⅰ),Aβ1-42and shank protein in hippocampal brain slices of each group were detected with Western blot.Results The results of HE staining showed that the arrangement of hippocampal neurons were disordered in the model group,with death and depletion of neurons.The arrangement and depletion of hippocampal neurons in each Rg1 group were less severe compared with the model group,with most significant improvement seen in the high-concentration Rg1 group.The results of transmission electron microscopy showed that the number of autophagosomes in brain slices in the model group was significantly higher than that in the blank control group,while each Rg1 group had fewer autophagosomes than the model group.The results of Western blot showed that,compared with the blank control group,levels of Shank1,P62 and LC3-Ⅰproteins in brain slices were decreased(all P<0.

关 键 词：阿尔茨海默病 人参皂苷类 自噬 

分 类 号：R-332[医药卫生] R285.5