基于代谢网络技术的补阳还五汤对APP/PS1双转基因小鼠干预作用的研究  

Study on the intervention effect of Buyang Huanwu Decoction on APP/PS1 double transgenic mice based on metabolic network technology

作　　者：汪白云[1] 杨柳 刘国良[2] 朴成玉 李庆伟 薛慧 刘斌[2] WANG Baiyun;YANG Liu;LIU Guoliang;PIAO Chengyu;LI Qingwei;XUE Hui;LIU Bin(Department of Pharmacy, Jinzhou Central Hospital, Liaoning Province, Jinzhou 121000, China;Jiamusi College, Heilongjiang University of Chinese Medicine, Heilongjiang Province, Jiamusi 154007, China;Center for Drug Safety Evaluation, Heilongjiang University of Chinese Medicine, Heilongjiang Province, Harbin 150040, China)

机构地区：[1]辽宁省锦州市中心医院药剂科,辽宁锦州121000 [2]黑龙江中医药大学佳木斯学院,黑龙江佳木斯154007 [3]黑龙江中医药大学药物安全性评价中心,黑龙江哈尔滨150040

出　　处：《中国医药导报》2019年第8期13-17,共5页China Medical Herald

基　　金：黑龙江省自然科学基金项目(H201318);黑龙江省教育厅科学技术研究项目(12521518);黑龙江中医药大学新药研究基金项目(2017xy04)

摘　　要：目的基于代谢靶标角度揭示补阳还五汤对APP/PS1双转基因小鼠的调节机制。方法选取20只7月龄APP/PS1双转基因小鼠为研究对象,建立阿尔茨海默病模型。将APP/PS1双转基因小鼠随机分为2组,即模型组和补阳还五汤组(9.26 g/kg);另选取10只健康的BALB/c小鼠作为空白组。补阳还五汤组灌胃给予补阳还五汤,空白组及模型组给予生理盐水。连续灌胃35 d后进行Morris水迷宫实验,测定小鼠学习记忆能力。在此基础上,采用高通量液相色谱质谱联用技术对各组小鼠尿液进行全面分析,通过终端代谢产物回溯疾病发展过程中的关键代谢酶或代谢通路,聚焦APP/PS1双转基因小鼠的发病机制以及补阳还五汤的作用机制。结果 Morris水迷宫实验中,与空白组比较,模型组穿越平台次数明显减少(P <0.01),补阳还五汤组穿越平台的次数显著增加(P <0.01)、在目标象限的停留时间显著增加(P <0.01)。空间定位航行实验中,补阳还五汤组较模型组的逃避潜伏期明显缩短(P <0.01)。通过进一步尿液代谢轨迹分析发现,各组小鼠尿液聚类良好,空白组和模型组间分离明显,补阳还五汤组则处于两组间。通过分组贡献较大的差异离子分析发现其主要分布在鞘脂代谢和脂肪酸代谢。给予补阳还五汤后上述代谢通路明显呈回调趋势。经高分辨质谱鉴定获取了4个代谢异常的生物标志物,分别是花生四烯酸、神经胺、L-棕榈酰肉毒碱、甘油磷脂酰胆碱。通过组学数据处理平台Metaboanalyst 4.0的在线分析可知其主要涉及两种代谢通路,分别是鞘脂代谢和花生四烯酸代谢。结论补阳还五汤对阿尔茨海默病小鼠作用明显,可有效改善其学习记忆能为。调节氨基酸代谢及能量转化可能是其调节阿尔茨海默病的关键机制。Objective To reveal the regulation mechanism of Buyang Huanwu Decoction on APP/PS1 double transgenic mice based on metabolomics. Methods An animal model of Alzheimer′s disease was replicated in 20 APP/PS1 double transgenic mice. APP/PS1 double transgenic mice were randomly divided into two groups: model group and Buyang Huanwu Decoction group (9.26 g/kg). Ten healthy BALB/c mice were selected as control group. Buyang Huanwu Decoction group orally administrated with Buyang Huanwu Decoction, control group and model group were given the same volume of normal saline. Water maze behavior part tested the learning and memory ability of mice after 35 days′ intragastric administration. Then, the urine metabolites profiling analysis was conducted based on the high-throughput liquid chromatography mass spectrometry. The key metabolic enzymes or metabolic pathways in the course of disease development were traced by terminal metabolites, focusing on the pathogenesis of APP/PS1 double transgenic mice and the mechanism of Buyang Huanwu Decoction. Results In the Morris space exploration experiment, compared with the blank group, the number of platform crossing in the model group was significantly reduced (P < 0.05), the number of platform crossing in the Buyang Huanwu Decoction group was significantly increased (P < 0.05), and the residence time in the target quadrant was significantly increased (P < 0.05). In the space positioning navigation experiment, the escape latency of Buyang Huanwu Decoction group was significantly shorter than that of model group (P < 0.05). Through further analysis of urine metabolic trajectory, it was found that the urine of mice in each group was clustered well, and the separation between blank group and model group was obvious, while Buyang Huanwu Decoction group was located between the two groups. Through the contribution analysis of larger difference ion, it was found that lipid metabolism and lipid metabolism were mainly distributed in sphingolipids. The above metabolic pathways showed a sign

关 键 词：补阳还五汤 阿尔茨海默病 代谢组学 通路 机制 

分 类 号：R289.5[医药卫生—方剂学]