基于动物实验和网络药理学的肺力咳合剂止咳、祛痰、平喘的作用研究

Research on function of Feilike Mixture in stopping cough, eliminating phlegm and relieving asthma based on animal experiments and network pharmacology

通过动物实验观察肺力咳合剂止咳、祛痰和平喘的药理作用,并结合网络药理学探讨其作用机制。采用豚鼠枸橼酸引咳实验观察止咳作用;小鼠气管酚红排泌实验、脂多糖致大鼠黏液高分泌实验观察祛痰作用;豚鼠磷酸组胺引喘实验观察平喘作用。采用TCMSP、TCMID、TCMIP、BATMAN-TCM数据库及文献查找肺力咳合剂各中药化学成分,并通过ADMETlab 2.0进行潜在活性成分的筛选。利用STITCH、SwissTargetPrediction和TCMSP进行成分靶点预测。通过SymMap数据库查询咳、痰、喘相关症状群靶点。将药物靶点与症状群靶点取交集后,分别利用OECloud工具进行GO生物过程分析和KEGG通路富集分析。结果显示,肺力咳合剂0.43~1.74 g·kg^(-1)可显著降低豚鼠3 min内咳嗽次数(P<0.05,P<0.01),6~12 g·kg^(-1)可增加小鼠气管酚红排泌量(P<0.01),2~8 g·kg^(-1)可抑制杯状细胞的数量(P<0.05,P<0.01);7~11.2 g·kg^(-1)可明显延长引喘潜伏期(P<0.05)。通过网络药理学分析,肺力咳合剂共获得115个潜在活性化学成分、910个靶点。肺力咳合剂止咳、祛痰、平喘交集靶点分别为27、12、7个,GO生物过程和KEGG通路结果发现三者存在共性和特性,其中细胞因子-细胞因子受体相互作用、感染性疾病相关信号通路是三者共同KEGG通路。通过实验验证和网络药理学分析均证明肺力咳合剂具有较好的止咳、祛痰和平喘作用。

This study observed the pharmacological effects of Feilike Mixture(FLKM) in stopping cough, eliminating phlegm, and relieving asthma through animal experiments, and explored its mechanism using network pharmacology. The antitussive effect was detected by citric acid-induced guinea pig cough model, the expectorant effect by mouse phenol red excretion experiment and lipopolysaccharide-induced mucus hypersecretion rat model, and the antiasthmatic effect by histamine phosphate-induced guinea pig asthma model. The chemical components of FLKM were collected by TCMSP, TCMID, TCMIP, and BATMAN-TCM databases and literature search, and the potential active components were screened through ADMETlab 2.0. The targets of FLKM were obtained by STITCH, SwissTargetPrediction, and TCMSP, and the symptom targets of cough, phlegm, and asthma were acquired through SymMap database. After taking the intersection of FLKM targets and symptom targets, this study used the OECloud tool to perform Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. Results demonstrated that FLKM 0.43-1.74 g·kgreduced the number of coughs in guinea pigs within 3 min(P<0.05, P<0.01), and FLKM 6-12 g·kgincreased the tracheal phenol red excretion in mice(P<0.01). Moreover, FLKM 2-8 g·kginhibited the number of goblet cells(P<0.05, P<0.01), and FLKM 7-11.2 g·kgprolonged the incubation period of asthma(P<0.05). A total of 115 potential active components and 910 targets of FLKM were obtained through network pharmacological analysis. FLKM had 27, 12, and 7 targets for stopping cough, eliminating phlegm, and relieving asthma, respectively. The GO and KEGG enrichment analysis found that there were commonalities and characteristics, among which cytokine-cytokine receptor interaction and infectious disease-related signaling pathway were shared. FLKM has a good effect of stopping cough, eliminating phlegm, and relieving asthma through animal experiments and network pharmacology.