二至丸对去卵巢所致AD小鼠海马蛋白质组学的影响

Effect of Erzhiwan on Hippocampus Proteomics of Mice of Ovariectomized-induced AD

目的：应用蛋白质组学方法探究二至丸对去卵巢所致阿尔茨海默病（Alzheimer’s disease，AD）小鼠的生物学基础的影响。方法：小鼠去卵巢后随机分为4组，分别为模型组、阳性药组、二至丸高、低剂量组，每组14只小鼠，另设14只为假手术组，术后6周开始给药，二至丸高、低剂量组分别用对应药物灌胃给药（2，1g·kg^-1）；阳性药组隔天皮下注射苯甲酸雌二醇0．02mg；假手术组与模型组灌胃等容量的蒸馏水，每日1次，持续给药32d。取小鼠海马组织，提取蛋白，Nan01．ESI液相．质谱联用系统检测蛋白，Protein Discovery软件鉴定蛋白，SIEVE软件对海马蛋白进行相对定量定性分析。PANTHER ClassificationSystem对差异蛋白进行基因本体（GO）分析，京都基因与基因组百科全书（KEGG）富集信号通路。结果：与模型组相比较，正常组、二至丸高、低剂量组小鼠共有80多个差异表达蛋白，通过GO分析分析发现这些蛋白可分为微管蛋白、热休克蛋白、能量代谢相关蛋白和脑保护相关蛋白等与AD相关的蛋白；KEGG分析发现以上差异蛋白共涉及20条信号通路，以上差异蛋白可能是二至丸防治AD的靶点蛋白。结论：二至丸可能通过促进能量代谢与囊泡运输，同时减少邶的生成与神经细胞的氧化损伤来达到治疗AD的作用。

Objective ： To explore the effect of Erzhiwan on the biological basis of mice of ovariectomized- induced Alzheimer＇s disease （AD） by proteomic method. Method： Ovariectomized mice were randomly divided into model group, positive control group, high-dose Erzhiwan group and low-dose Erzhiwan group, with 14 mice in each group. Another 14 mice were included in the sham operation group. Six weeks later after the operation, the mice in the sham operation group and the model group were given the same volume of distilled water, high and low-dose Erzhiwan groups were given the corresponding drugs by gavage, with the drug concentration of 2, 1 g＇kg-~, and the positive control group was injected with estradiol benzoate 0.02 mg every other day. The drug administration lasted for 32 days. Proteins were extracted from the mice＇s hippocampus, and detected by Nanol-ESI liquid-mass spectrometry system, and then identified by Protein Discovery software; and a quantitative analysis was made for the hippocampal proteins by SIEVE software. PANTHER Classification System was used for gene ontology （GO） analysis of differential proteins, and Kyoto Encyclopedia of Genes and Genomes （KEGG） was used to enrich signal pathways. Result： Compared with the model group, there were more than 80 differentially expressed proteins in the normal group and the high and low-dose Erzhiwan groups. Through the GO analysis, these proteins could be divided into tubulin, heat shock protein, energy metabolism-related protein, brain protection-related proteins and other AD-related proteins. KEGG analysis found that the above differential proteins involved 20 signal pathways. These differential proteins may be the target for Erzhiwan in preventing and treating AD. Conclusion： Erzhiwan can promote the energy metabolism and vesicle transport, while reducing the production of A- and the oxidative neurons, so as to achieve the therapeutic effect damage ofneurons, so as to achieve the therapeutic effect of AD.