Title:Methodological Verification-based Screening of the Representative Ingredients
for Traditional Chinese Medicine: Taking Astragalus as an Example
for Interfering with Cervical Cancer
Volume: 18
Issue: 5
Author(s): Hao Sun*, Dan Wang, Mengjin Xu, Yi Gao and Fan Li
Affiliation:
- Department of Pharmacy, Women's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China
Keywords:
Astragalus, cervical cancer, representative ingredients, disease network clusters, methodological verification, in vitro cell experiments.
Abstract:
Background: The screening of effective ingredients is the bridge between the research of
efficacy and the mechanism of traditional Chinese medicine. Although promising virtual screening has
emerged as an attractive alternative, an ideal strategy is still urgently required due to the characteristics
of multi-ingredients and multi-targets of traditional Chinese medicine.
Objective: The aim of the study was to develop a methodological verification-based novel screening
strategy capable of comprehensively assessing the ability of compounds to perturb disease networks,
thereby identifying representative ingredients of traditional Chinese medicine interventions in complex
diseases.
Methods: In this article, we take astragalus interfering with cervical cancer as an example. First, a multifunctional
clustering disease network model was constructed; second, the several drugs and their decoys
were used for molecular docking with disease network clusters for methodological verification
and determining the best scoring criteria. Third, the representative ingredients of astragalus were
screened according to the best scoring criteria. Finally, the effects of the representative ingredients on
cervical cancer SiHa cells were evaluated by CCK-8 assay, flow cytometry, and western blot analysis.
Results: Three representative ingredients of astragalus were betulinic acid, hederagenin and methylnissolin,
which perturbed the apoptosis, stabilization of p53, and G1/S transition cluster as a whole, respectively.
CCK-8 assay showed that the IC50 value of betulinic acid, hederagenin and methylnissolin
at 48 h was 28.84, 101.90, and 187.40 μM, respectively. Flow cytometry showed that these three representative
ingredients could significantly induce early apoptosis and cell cycle arrest. Western blot analysis
showed that betulinic acid treatment significantly increased p53 expression, while hederagenin and
methylnissolin did not.
Conclusion: This study has provided new ideas for the screening of effective ingredients in traditional
Chinese medicine, and established a foundation for elucidating the overall mechanism of action of traditional
Chinese medicine.
