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目的:参附注射液是传统中药红参与附子的复方制剂,临床主适应症为充血性心力衰竭、缺血性脑卒中等。以缺血性脑卒中为导向,采用系统生物学和实验研究方法,对参附注射液调节生物学网络的分子机制做一阐释。方法:以参附注射液的28个成分出发,分别从TCMGene DIT数据库系统和Agilent literature search系统中挖掘参附注射液中成分作用的蛋白质数据,并辅助以Pharmmapper反向对接靶标,构建参附注射液多成分-蛋白网络,在Genecards,BIND,Bio GRID,Int Act,Mint等数据库中挖掘蛋白之间关联,建立蛋白相互作用网络。提取显著性差异蛋白子网络一个,并对其中通路以Western blot蛋白印迹方法加以验证。结果:参附注射液成分与55个蛋白有连接,构成53个无孤立结点的蛋白相互作用网络。应用Cluster One模块对蛋白相互作用网络富集分析,提取显著性差异P<0.05的子网络1个,子网络中含有关键蛋白15个,经Biocart信号通路映射,涉及NF-κB信号通路,AKT信号通路,Toll样受体信号通路,MAPK信号通路等。采取Western blot方法对富集指数P值最小的NF-κB信号通路进行验证,发现参附注射液在缺血1 h再灌注24 h的MCAO模型上对NF-κB p65和IκB-α的磷酸化表达有明显下调作用。结论:采用计算机系统生物学和实验验证方式初步阐释了参附注射液主要化合物防治疾病的分子机制,并以分子生物学实验方式对所预测的信号通路进行验证,为中药复方的系统研究提供参考。
OBJECTIVE: Shenfu injection is a traditional Chinese medicine red involved in the aconite compound preparation, the main clinical indications for congestive heart failure, ischemic stroke. To ischemic stroke-oriented, using the system biology and experimental research methods, the molecular mechanism of Shenfu injection regulating biological networks to make an explanation. METHODS: Based on the 28 components of Shenfu injection, the protein data of the components of Shenfu injection were excised from the TCMGene DIT database system and the Agilent literature search system, respectively, and the reverse docking of the target with the Pharmmapper was performed. Liquid multi-component-protein network to discover the protein-protein interaction network in Genecards, BIND, Bio GRID, Int Act, Mint and other databases. One of the distinct protein sub-networks was extracted, and the pathway was verified by Western blot. Results: The composition of Shenfu injection was connected with 55 proteins, forming 53 protein-protein interaction networks without isolated nodes. Using Cluster One module to enrich and analyze the protein interaction network, one sub-network with significant difference P <0.05 was extracted and 15 key proteins in the sub-network were obtained. The Biocart signal pathways involved in NF-κB signal pathway, AKT signal Pathway, Toll-like receptor signaling pathway, MAPK signaling pathway and the like. Western blot was used to verify the NF-κB signaling pathway with the lowest enrichment index (P value). The results showed that Shenfu injection on NF-κB p65 and IκB-α phosphorylation Significantly down-regulated expression. CONCLUSION: The molecular mechanism of Shenfu injection’s main compounds in prevention and treatment of diseases was preliminary elucidated by means of computer system biology and experimental verification. The predicted signal pathways were verified by molecular biological experiments to provide reference for the systematic study of traditional Chinese medicine compounds .