基于海马区乙酰化蛋白组学探讨原络通经针法对AD模型小鼠记忆损伤的影响  

To investigate the effect of Yuanluo Tongjing acupuncture on memory impairment of AD model mice based on hippocampal acetylated proteomics

作　　者：邢菁[1] 徐佳[1] 史珊怡[1] 陈奥[1] 郭文海[1] XING Jing;XU Jia;SHI Shan-Yi(Prevention and Treatment Center of The Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin 150001,Heilongjiang,China)

机构地区：[1]黑龙江中医药大学附属第二医院治未病中心,黑龙江哈尔滨150001

出　　处：《吉林医学》2025年第1期4-9,共6页Jilin Medical Journal

基　　金：黑龙江省中医药科研项目[项目编号:ZHY2023-027];孙申田青年人才基金[项目编号:2021KY-16];黑龙江省中医药学会青年人才托举工程项目[项目编号:2022-QNRC1-26]。

摘　　要：目的:基于海马区乙酰化蛋白组学探讨原络通经针法对阿尔茨海默病(AD)模型小鼠记忆损伤的影响。方法:按照随机数字表法将30只健康雄性昆明种小鼠分为空白组、模型组和针刺组各10只,模型组和针刺组小鼠腹腔注射东莨菪碱溶液[100 mg/(kg·d)],空白组腹腔注射等量的生理盐水,持续注射7d造模。针刺组小鼠采用原络通经针法进行针刺(1次/d),空白组和模型组进行相同时间和程度的捉抓,连续干预30 d。采用Morris水迷宫行为学实验和新物体识别实验检测小鼠记忆能力,采用HE染色观察小鼠海马组织病理损伤情况,采用液相色谱-串联质谱法(LC MS/MS)检测小鼠海马组织中乙酰化修饰蛋白,并对其进行生物学信息分析。结果:与模型组比较,针刺组的逃避潜伏期、逃逸路径长度均明显缩短,第2象限停留时间明显增长,通过原平台位置次数明显增多,识别指数明显增高,差异有统计学意义(P<0.05)。与模型组比较,针刺组小鼠海马组织的病理损伤明显减轻,凋亡细胞明显减少。LC MS/MS分析显示,与空白组及模型组比较,针刺组乙酰化修饰整体水平增高。通过乙酰化蛋白相互作用网络分析筛选出5个关键枢纽蛋白为Hspa8、Rab7a、Nsf、Ezr、Cfl1,模型组中该5个关键枢纽蛋白无乙酰化修饰,而针刺组该5个关键枢纽蛋白发生乙酰化修饰。结论:原络通经针法能有效改善AD模型小鼠记忆损伤,减轻海马区病理损害,可能与海马区关键枢纽蛋白乙酰化修饰有关。Objective To investigate the effect of Yuanluo Tongjing acupuncture on memory impairment of Alzheimer′s disease(AD)model mice based on hippocampal acetylated proteomics.Method 30 cases of healthy male Kunming breed mice were divided into blank group,model group and acupuncture group with 10 mice in each group according to random number table method.Mice were intraperitoneally injected with scopolamine solution[100 mg/(kg·d)]in model group and acupuncture group,while mice were intraperitoneally injected with the same amount of normal saline in blank group,continuous injection for 7 days to moldel.The mice were needled with the original collateral-Tongjing acupuncture method(once/d)in the acupuncture group,the mice were caught at the same time and degree in the blank group and the model group were,and the intervention lasted for 30 days.Morris water maze behavior test and new object recognition test were used to detect the memory ability of mice.HE staining was used to observe the pathological injury of mice hippocampus.The liquid chromatography-tandem mass spectrometry(LC MS/MS)was used to determine acetylated modified proteins from mouse hippocampal tissues and analyze their biological information.Results Compared with the model group,the escape latency period and escape path length of the acupuncture group were significantly shortened,the residence time in the second quadrant was significantly increased,the number of times through the original platform position was significantly increased,and the recognition index was significantly increased in the acupuncture group(P<0.05).Compared with the model group,the pathological damage and apoptosis of hippocampus in the acupuncture group were significantly reduced in the acupuncture group.LC MS/MS analysis showed that compared with blank group and model group,the overall level of acetylation modification increased in the acupuncture group.Five key hub proteins were identified by acetylation protein interaction network analysis as Hspa8,Rab7a,Nsf,Ezr and Cfl1.The five

关 键 词：原络通经针法 阿尔茨海默病 小鼠 乙酰化蛋白组学 记忆损伤 

分 类 号：R749[医药卫生—神经病学与精神病学]