基于动物实验和网络药理学探讨复方伤痛胶囊治疗股骨干骨折的作用机制

Mechanism of Fufang Shangtong Capsule in the treatment of femoral shaft fracture based on animal experiment and network pharmacology

关键词:  股骨干骨折  复方伤痛胶囊  网络药理学

Key words:  femoral shaft fracture  Fufang Shangtong Capsule  network pharmacology

摘要:

[目的] 基于动物实验和网络药理学技术探讨复方伤痛胶囊治疗股骨干骨折的潜在作用机制。[方法] 构建股骨干骨折大鼠模型并观察复方伤痛胶囊干预后的微计算机断层扫描（Micro-CT）结果、生物力学分析结果、血清白细胞介素-6（IL-6）水平、血清肿瘤坏死因子-α（TNF-α）水平和血管内皮生长因子（VEGF） mRNA阳性细胞数量。通过中药系统药理学数据库与分析平台（TCMSP）数据库检索出药物活性成分和作用靶点。在在线人类孟德尔遗传数据库（OMIM）和GeneCards数据库中检索与股骨干骨折相关的疾病靶点，将其与药物靶点取交集，作为复方伤痛胶囊治疗股骨干骨折的作用靶点。利用Cytoscape软件构建“药物-成分-作用靶点”网络图，在String数据库中得到靶点蛋白质相互作用（PPI）网络关系，使用Metascape数据库对作用靶点进行生物信息学分析。[结果] 动物实验结果表明，复方伤痛胶囊治疗组大鼠在Micro-CT扫描下显示骨折端恢复良好，生物力学检测结果显示治疗组大鼠股骨干最大载荷高于对照组。复方伤痛胶囊可下调股骨干骨折大鼠IL-6、TNF-α的表达，上调VEGF的表达。在网络药理学研究结果中获取复方伤痛胶囊209种成分及60个治疗股骨干骨折的靶点，PPI网络中自由度靠前的5个靶点是白蛋白（ALB）、蛋白激酶Ｂα（AKT1）、IL-6、TP53和VEGFA，基因本体（GO）富集分析共富集到452个GO条目，京都基因组百科全书（KEGG）富集分析得到与治疗股骨干骨折相关的通路有23条。[结论] 本研究初步阐明了复方伤痛胶囊多成分、多靶点、多途径治疗股骨干骨折的作用机制，为其进一步开发利用提供依据。

Abstract:

[Objective] To explore the potential mechanism of Fufang Shangtong Capsule in the treatment of femoral shaft fracture based on animal experiment and network pharmacology. [Methods] The rat model of femoral shaft fracture was established,and the Micro-CT results,biomechanical analysis results,serum IL-6 levels,serum TNF-α levels and the number of VEGF mRNA positive cells were observed after taking Fufang Shangtong Capsule. The active ingredients and targets of drugs of Fufang Shangtong Capsule were screened by TCMSP database. The related targets of Femoral Shaft Fracture were searched by Genecards and OMIM databases,and their intersection with the targets of drugs were selected as potential targets of Fufang Shangtong Capsule for the treatment of Femoral Shaft Fracture. The visual "drug-ingredient-target" network was constructed by Cytoscape. The protein-protein interaction(PPI) network was obtained by the String database. The bioinformatics analysis were carried out for these potential targets by the Metascape database. [Results] Animal experiment indicated that the rats in the Fufang Shangtong Capsule treatment group recovered well under Micro-CT scanning,and the biomechanical test results showed that the maximum load of the femoral shaft in the treatment group was higher than that in the control group. Fufang Shangtong Capsule can reduce the expression of IL-6 and TNF-α,and increase the expression of VEGF in rats with femoral shaft fracture. The 209 active ingredients and 60 potential targets for the treatment of femoral shaft fracture were obtained from the Fufang Shangtong Capsule in the research results of network pharmacology. The top 5 target genes with degree in the PPI network were ALB,AKT1,IL-6,TP53 and VEGFA. Gene Ontology(GO) enrichment analysis enriched a total of 452 GO entries,and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis yielded 23 pathways associated with the treatment of femoral stem fractures. [Conclusion] This study preliminarily elucidates the mechanism of Fufang Shangtong Capsule in the treatment of femoral shaft fracture with multi-components,multi-targets and multi-pathways to provide a basis for its further development and utilization.