太子参多糖-葛根淫羊藿总黄酮对小鼠脑神经蛋白质组学作用的比较

Comparison Study of the Neuroproteomic Effects of Polysaccharides of Radix Pseudostellariae and the Flavonoids of Radix Epimedii and Radix Pueraria on the Brain of Mice

目的观察太子参多糖(总多糖)、葛根淫羊藿总黄酮(总黄酮)及其复合物对小鼠脑蛋白质组学的影响,探讨其神经细胞网络调控的潜在机理。方法给正常小鼠分别灌胃太子参多糖(400 mg·kg^(-1))、葛根淫羊藿总黄酮(400 mg·kg^(-1))及其二者复合物(800 mg·kg^(-1)),连续给药7天,第7天最后1次给药后1 h,处死动物,取脑。提取蛋白,利用LC/MS-MS测试蛋白表达的变化。通过DAVID数据库分析变化蛋白的功能及信号通路,STRING数据库分析变化蛋白的相互作用及其网络关系。结果在所测试的5567个蛋白中,以上调蛋白占比较高,其中主要是线粒体的基因受到调控。总多糖、总黄酮及其复合物都上调的蛋白有8个,作用于线粒体的不同环节,提示该复合物抗抑郁作用的靶点主要在线粒体。太子参多糖的调控线粒体相关蛋白的作用揭示了其潜在的神经精神方面的药理作用。除此,还发现各有效部位调控蛋白的数量明显多于复合物,提示药物复合作用的复杂性。结论总多糖、总黄酮及其复合物可以作用于脑神经元,并且主要以线粒体为主要靶点。该研究为相关中药调控神经精神功能活动提供了重要的蛋白线索及其实验依据。

Objective To observe the proteomic profiling of mouse brain after oral administration of(total polysaccharides),the total flavonoids of Radix Epimedii and Radix Pueraria(total flavonoids)and their mixture complexes(complexes),and to explore the underlying mechanism of the protein network of neuron.Methods The normal mice were orally given total polysaccharides(400 mg·kg^(-1)),total flavonoids(400 mg·kg^(-1))and their complexes(800 mg·kg^(-1))for 7 consecutive days.On the 7th day one hour after the last administration,the mice were sacrificed and brains were isolated.After extracting the brain protein,the samples were detected useing LC/MS-MS observing the change of protein expression.Through the DAVID database,the function annotation and signaling pathway of the proteins are clustered,and the interaction and network relationship of the changed proteins were analyzed by the STRING database.Results Among the 5567 proteins tested,up-regulated proteins accounted for a relatively high proportion,mainly mitochondrial genes were regulated.Among them,there are 8 up-regulated proteins occured in total polysaccharides,total flavonoids and their complexes,which act on different sites of mitochondria,suggesting that the anti-depressant effect of the drug is mainly in the mitochondria.Notably,the role of total polysaccharides in regulating mitochondrial-related proteins reveals its potential neuropsychological pharmacological effects presented by our research.In addition,it was found that the number of alternated proteins in each effective site was significantly greater than that of the complex,suggesting the complexity of drug mixture.Conclusion Total polysaccharides,total flavonoids and their complexes can act on brain neurons,and mainly target mitochondria.This study provides important protein clues and experimental basis for the Chinese herbs involved in regulating neuropsychiatric function.