基于网络药理学探讨温胆汤治疗血管性痴呆作用靶点和作用机制研究

To Explore the Target and Mechanism of Wendan Decoction in the Treatment of Vascular Dementia Based on Network Pharmacology

目的基于中药网络药理学和分子生物学探讨温胆汤(WDT)治疗血管性痴呆(vascular dementia,VD)的作用靶点及分子机制。方法应用BATMAN-TCM数据库及TCMSP数据库对温胆汤的活性成分及作用靶点进行筛选,在GeneCards、OMIM数据库中筛选VD的作用靶点,在通过Venn温恩图工具获取温胆汤与VD的交集核心靶点,再应用Cytoscape 3.7.2软件构建化合物-靶点-疾病网络,应用R3.6.0软件对交集靶点进行基因本体(gene ontology,GO)功能富集和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析;高脂模型组18只,高脂饲料连续喂养13周,在第9周制备大鼠VD模型(采用改良的双侧颈总动脉分次结扎的方法)。手术造模结束后,在实验组存活的大鼠中展开行为学筛选,在筛选后的12只大鼠中,随机分为两组,高脂+VD模型组(以下称模型组)和高脂VD模型+温胆汤组(以下称中药组),每组6只,假手术组6只。假手术组(以下称正常组)以普通饲料喂养,第10周,中药组给予温胆汤(3.62 g/kg),每日灌胃给药1次,持续4周,模型组每日灌胃等体积生理盐水,持续4周;采用RT-qPCR法检测大鼠海马组织Bcl-2、Bax、Cyt-c、Caspase3、Caspase9 mRNA相对表达水平。结果筛选出WDT中145种活性成分有233个VD的潜在治疗靶点,其中有181个靶点之间存在的4133种蛋白蛋白相互作用关系,WDT对线粒体凋亡的调控是其防治VD的重要生物学途径,预测其核心调控靶点有Bcl-2、Bax、Cytc、Caspase3、Caspase9等,动物实验RT-qPCR验证了预测结果,证明WDT可能通过促进Bcl-2基因表达,抑制Bax、Cytc、CASP9、CASP3基因表达治疗VD。结论WDT活性化合物对线粒体凋亡相关的多个靶点和多条信号通路的调控是其防治VD的重要作用机制。

Objective Based on the network pharmacology and molecular biology of traditional Chinese medicine,to explore the target and molecular mechanism of Wendan Decoction(WDT)in the treatment of vascular dementia(VD).Methods Comprehensive application of traditional Chinese medicine database(TCMSP)and BATMAN-TCM analysis tool select Wendan Tang,the active ingredient and targets,related to the disease target database search GeneCards,OMIM screening of VD targets,and then screening of Wendan Tang and VD intersection of core targets,and using Cytoscape 3.7.1 software build compound-targets-disease network,target to make use of R packages GO function of enrichment and enrichment of KEGG pathway analysis;Eighteen rats in the high fat model group were fed with high fat diet for 13 consecutive weeks.At the 9 th week,the VD model of rats was prepared by the method of the modified bilateral common carotid artery ligation.After modeling,the surviving rats in the experimental group were screened by behavior,and the 12 rats after screening were randomly divided into two groups:high fat+VD model group(hereinafter referred to as model group)and high fat VD model+Wendan decoction group(hereinafter referred to as traditional Chinese medicine group),with 6 rats in each group and 6 rats in the sham operation group.The sham operation group(hereinafter referred to as the normal group)was fed with ordinary feed.At week 10,the TCM group was given Wendan Decoction(3.62 g/kg)once a day for 4 weeks,while the model group was given the same volume of normal saline daily for 4 weeks.RT-qPCR was used to detect the mRNA relative expression levels of Bcl-2,Bax,Cyt-c,Caspase3 and Caspase9 in rat hippocampus.Results select 145 kinds of active ingredients in the WDT has 233 potential therapeutic targets for VD,there are 181 targets between 4133 egg albumin interaction relations,WDT of mitochondrial apoptosis regulation is one of the important biological pathways,its prevention and treatment of VD predict its core control targets of Bcl-2 and Bax,Cytc,Caspase3 and Caspase9,RT-qPCR of animal experiments verify the predicted results and the WDT to justify the possible by promoting Bcl-2 gene expression,inhibition of Bax,Cytc,Caspase3 and Caspase9 gene expression in the treatment of VD.Conclusions The regulation of WDT active compounds on multiple targets and multiple signaling pathways related to mitochondrial apoptosis is an important mechanism of WDT active compounds in the prevention and treatment of VD.