基于miR-124/SIRT1信号通路探讨二至丸对神经突触损伤小鼠的作用及机制  

Exploration of the Effect and Mechanism of Erzhi Pills on Synaptic Damage of Mice Based on miR-124/SIRT1 Signaling Pathway

作　　者：王紫琪 陈淑云 汪蕊 吕锦瀚 黄楚瑶 赵威[1] WANG Ziqi;CHEN Shuyun;WANG Rui;LYU Jinhan;HUANG Chuyao;ZHAO Wei(Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学科技创新中心,广东广州510405

出　　处：《中药新药与临床药理》2024年第12期1808-1815,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省自然科学基金面上项目(2019A1515011299)。

摘　　要：目的基于miR-124/SIRT1信号通路探讨二至丸对神经突触损伤小鼠的作用及机制。方法将C57BL/6J小鼠随机分为空白组、模型组、阴性病毒对照组、阳性药物组(白藜芦醇50mg·kg^(-1)·d^(-1))、二至丸组(3.5 g·kg^(-1)·d^(-1)),每组10只。采用脑室注射miR-124-UP腺相关病毒复制miR-124-UP小鼠模型。灌胃给药,每日1次,连续42 d。采用Morris水迷宫实验检测小鼠学习记忆能力;HE及Nissl染色法观察海马组织病理变化;检测血清丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-PX)水平;Western Blot法检测海马组织中SIRT1、BDNF、CREB、SYN、PSD95蛋白表达水平;qPCR法检测海马组织miR-124、SIRT1、BDNF、CREB、SYN、PSD95 mRNA表达水平。结果与空白组比较,模型组小鼠第5天的逃避潜伏期显著延长(P<0.01),穿越平台次数显著减少(P<0.001);小鼠海马CA3区神经细胞明显减少,细胞呈萎缩状态,细胞核浓染;血清MDA含量显著升高(P<0.001),GSH-PX酶活性显著下降(P<0.001);海马组织SIRT1、BDNF、CREB、SYN、PSD95蛋白及mRNA表达量均显著下降(P<0.05,P<0.01,P<0.001);海马组织miR-124 mRNA表达量显著升高(P<0.01)。与模型组比较,阳性药物组与二至丸组第5天的逃避潜伏期显著缩短(P<0.01),穿越平台次数显著增加(P<0.01,P<0.001);小鼠海马CA3区神经细胞数量增加,细胞形态趋于正常,萎缩细胞数量减少;血清MDA含量显著下降(P<0.001),GSH-PX酶活性显著升高(P<0.01,P<0.001);海马组织SIRT1、BDNF、CREB、PSD95蛋白表达量均显著升高(P<0.01,P<0.001);海马组织miR-124 mRNA表达量显著降低(P<0.01),SIRT1、BDNF、CREB、SYN、PSD95 mRNA表达量均显著升高(P<0.05,P<0.01,P<0.001);二至丸组小鼠海马组织SYN蛋白表达量显著升高(P<0.001)。结论二至丸能够提高miR-124-UP小鼠的学习记忆能力,增强抗氧化应激能力,改善神经元损伤,可能与其通过miR-124/SIRT1信号通路调控神经元可塑性有关。Objective To investigate the effect and mechanism of Erzhi Pills on synaptic damage of mice based on miR-124/SIRT1 signaling pathway.Methods C57BL/6J mice were randomly divided into blank group,model group,negative viral control group,positive drug group(resveratrol 50 mg·kg^(-1)·d^(-1)),and Erzhi Pills group(3.5 g·kg^(-1)·d^(-1)),with 10 mice in each group.A miR-124-UP mouse model was replicated using intraventricular injection of adenovirus.The corresponding drug was administered by gavage after modeling,once a day for 42 days.Morris water maze was used to detect the learning and memory ability of the mice.Pathological changes in hippocampal tissue were observed using HE and Nissl staining.The kits were used to detect the serum levels of malondialdehyde(MDA)and glutathione peroxidase(GSH-PX).Western Blot was used to determine the protein expression of SIRT1,BDNF,CREB,SYN and PSD95 in hippocampal tissues,and qPCR was used to determine the mRNA expression of miR-124,SIRT1,BDNF,CREB,SYN and PSD95 in hippocampal tissues.Results Compared with the blank group,escape latency at the 5th day of the model group was significantly prolonged(P<0.01),while the number of crossing platforms was significantly reduced(P<0.001).Nerve cells in the CA3 region of the hippocampus were markedly reduced,with atrophied cells and densely stained nuclei.Serum MDA content was markedly up-regulated(P<0.001),and GSH-PX enzyme activity appeared to be significantly down-regulated(P<0.001).The protein and mRNA expression of SIRT1,BDNF,CREB,SYN,and PSD95 in hippocampal tissue appeared significantly down-regulated(P<0.05,P<0.01,P<0.001),but the mRNA expression of miR-124 in hippocampal tissue was significantly up-regulated(P<0.01).Compared with the model group,escape latency at the 5th day of positive drug group and Erzhi Pills group was obviously shortened(P<0.01)and the number of crossing platforms was significantly increased(P<0.01,P<0.001).Nerve cells in the CA3 region of the hippocampus was increased,the cell morphology tended to be norm

关 键 词：二至丸 神经退行性改变 突触损伤 miR-124/SIRT1信号通路 神经元可塑性 小鼠 

分 类 号：R285.5[医药卫生—中药学]