Network pharmacology and experimental validation to explore the molecular mechanisms of Bushen Huoxue for the treatment of premature ovarian insufficiency

Bioengineered. 2021 Dec;12(2):10345-10362. doi: 10.1080/21655979.2021.1996317.

Abstract

Bushen Huoxue (BSHX) has been applied in clinical traditional Chinese medicine treatment, and has definitive clinical efficacy in the treatment of Premature Ovarian Insufficiency (POI) in China. However, little is known of the underlying mechanism of BSHX. The purpose of this paper is to study the pharmacological mechanisms of BSHX acting on POI based on a pharmacology and experimental validation. The pharmacological database of chinese medicine system and analysis platform (TCMSP) were used to search the effective active ingredients and potential action targets of BSHX. Drugbank, Online Mendelian Inheritance in Man (OMIM), Genecards, and Disgenet databases were used to obtain relevant targets of POI. Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and the visual network of protein-protein interaction network were constructed by FunRich3.1. Pymol software, and Auto Dock tools 1.5.6 were used for molecular docking. Murine model of POI was used to further investigate the mechanism of BSHX against on POI. Finally, 127 active compounds were collected from TCMSP database, and 215 active targets were identified. There were 1366 targets related to POI and 99 targets of BSHX for the treatment of POI. Quercetin, kaempferol, and stigmasterol were recognized as the most effective compounds corresponding to targets. The top three genes according to degree value are TP53, Akt1, and VEGFA. Further, the results of GO and KEGG enrichment analysis revealed that those core targets were mainly enriched on TRAIL and TGF-β receptor signaling. The results of molecular docking showed that stigmasterol had good binding ability to Akt1. Moreover, experimental validation suggests that BSHX significantly Increased the expression of TGF-β1 and Smad2/3, regulating the release of serum sex hormones, which include Follicular stimulating hormone (FSH), Estradiol (E2), and Antimullerin hormone (AMH).

Keywords: Bushen Huoxue; experimental validation; molecular docking; molecular mechanism; network pharmacology; premature ovarian insufficiency.

Plain language summary

HIGHLIGHTSBSHX treats POI by regulating TGF-β1 and Smad2/3 signaling pathways; Quercetin, kaempferol, and stigmasterol were the most effective compounds in BSHX.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Administration, Oral
  • Animals
  • Drugs, Chinese Herbal / administration & dosage
  • Drugs, Chinese Herbal / pharmacology
  • Drugs, Chinese Herbal / therapeutic use*
  • Female
  • Gene Expression Regulation
  • Gene Ontology
  • Mice, Inbred ICR
  • Molecular Docking Simulation
  • Network Pharmacology*
  • Primary Ovarian Insufficiency / drug therapy*
  • Primary Ovarian Insufficiency / genetics
  • Primary Ovarian Insufficiency / pathology
  • Protein Interaction Maps / drug effects
  • Reproducibility of Results
  • Smad Proteins / metabolism
  • Thermodynamics
  • Transforming Growth Factor beta1 / metabolism

Substances

  • Drugs, Chinese Herbal
  • Smad Proteins
  • Transforming Growth Factor beta1
  • bushen huoxue

Grant support

This study was supported by the National Natural Science Foundation of China (grant no. 82074487, 81903998), Science and Technology Support Program of Jiangsu Province (grant no. BE2019766). Graduate Student training innovation project in Jiangsu Province (grant no. SJCX21-0754).