丹酚酸A对缺血性脑卒中模型大鼠海马区蛋白影响的串联质谱标签蛋白组学分析  

Salvianolic acid A effects on hippocampal protein expression in ischemic stroke rats:a tandem mass tag-based proteomic analysis

作　　者：许雯珊 江萍利 刘雨露 丁妍怡 余燕 杨敏光 柳维林[2,3] 陈立典 Xu Wenshan;Jiang Pingli;Liu Yulu;Ding Yanyi;Yu Yan;Yang Minguang;Liu Weilin;Chen Lidian(College of Rehabilitation Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;Academy of Rehabilitation Industry,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;National&Local Joint Engineering Research Center for Rehabilitation Medical Technology,Fuzhou 350122,Fujian Province,China)

机构地区：[1]福建中医药大学康复医学院,福建福州350122 [2]福建中医药大学康复产业研究院,福建福州350122 [3]康复医疗技术国家地方联合工程研究中心,福建福州350122

出　　处：《中国组织工程研究》2022年第23期3714-3720,共7页Chinese Journal of Tissue Engineering Research

基　　金：中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目”(2060302),项目负责人:柳维林。

摘　　要：背景:丹酚酸A对缺血性脑卒中后缺血侧海马区蛋白表达的影响及其作用机制尚未明确。目的:观察丹酚酸A对缺血性脑卒中大鼠海马区蛋白差异表达的影响,探讨其可能的神经保护机制。方法:采用改良Longa方法制备缺血性脑卒中大鼠模型,造模后随机分为生理盐水组与丹酚酸A组(n=6)。丹酚酸A组于线栓拔出后尾静脉注射2 mg/kg丹酚酸A;生理盐水组注射等体积的生理盐水。采用改良神经功能缺损评分标准评价大鼠神经功能;T2加权成像检测脑梗死体积;旷场实验及Morris水迷宫测试观察各组大鼠的自主行为、学习记忆能力;串联质谱标签定量蛋白组学技术分析缺血性脑卒中模型大鼠缺血侧海马区蛋白的差异表达。结果与结论:①与生理盐水组相比,丹酚酸A组大鼠改良神经功能缺损评分显著降低;②T2加权成像显示,丹酚酸A组较生理盐水组脑梗死体积明显减少;③旷场实验结果发现,丹酚酸A组大鼠自主活动及自由探索行为表现更佳;④Morris水迷宫测试结果表明,丹酚酸A组大鼠在定向航行实验中逃避潜伏期明显缩短、穿越平台次数相较于生理盐水组显著增加;⑤缺血侧海马区差异蛋白质分析结果发现,丹酚酸A组与生理盐水组之间筛选出50个上调及3个下调差异蛋白;基因本体论分析表明它们主要参与binding,catalytic activity等分子功能;京都基因和基因组百科全书信号通路分析显示,它们主要涉及Complement and coagulation cascades,Staphylococus aureus infection等信号通路;⑥结果显示,丹酚酸A可能通过调控缺血性脑卒中大鼠缺血侧海马区差异蛋白、补体及凝血级联信号通路,对缺血性脑卒中大鼠发挥神经保护作用。BACKGROUND:It is unclear about the effect of salvianolic acid A on hippocampal protein expression after ischemic stroke and its mechanism.OBJECTIVE:To observe the effect of salvianolic acid A on differential protein expression in the hippocampus of ischemic stroke rats,and to explore its possible neuroprotective mechanism.METHODS:The modified Longa method was used to prepare a rat model of ischemic stroke.After modeling,the rats were randomly divided into normal saline group(n=6)and salvianolic acid A group(n=6).Salvianolic acid A(2 mg/kg)was injected through the tail vein after the cord plug was pulled out.The normal saline group was injected with the equal volume of normal saline.Neurological function of the rats was evaluated by the modified neurological deficit score.T2-weighted imaging was used to detect infarct volume.Open field test and Morris water maze test were used to observe the autonomous behavior,learning and memory ability of rats in each group.Tandem mass tag-based quantitative proteomics technique was used to analyze differential protein expression in the ischemic hippocampus of ischemic stroke model rats.RESULTS AND CONCLUSION:Compared with the normal saline group,the modified neurological deficit score of rats was significantly decreased in the salvianolic acid A group.T2-weighted imaging results showed that the cerebral infarction volume in the salvianolic acid A group was significantly lower than that in the normal saline group.Results from the open field test showed that the rats in the salvianolic acid A group had better performance of autonomous activity and free exploration.Morris water maze test results showed that in the directional navigation experiment,the escape latency was significantly shortened and the number of platform crossings was significantly increased in the salvianolic acid A group compared with the normal saline group.Analysis of differentially expressed proteins in the ischemic hippocampus revealed that 50 differentially expressed proteins were up-regulated and 3 differe

关 键 词：丹酚酸A 缺血性脑卒中 蛋白组学 串联质谱标签技术 神经保护 海马 差异蛋白 生物信息学 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]