Exploration on the Mechanism of Gufang Granules for the Treatment of Osteoporosis Based on Network Pharmacology,Molecular Dynamics Simulations and in Vitro Experimental Validation
10.19879/j.cnki.1005-5304.202408448
- VernacularTitle:基于网络药理学、分子动力学模拟和体外实验验证探讨骨方颗粒治疗骨质疏松症作用机制
- Author:
Xiaoqing CHEN
1
;
Yangling HUANG
;
Shanshan LI
;
Chunbo LIANG
;
Yunzhao GONG
;
Wei CHEN
Author Information
1. 辽宁中医药大学,辽宁 沈阳 110847
- Publication Type:Journal Article
- Keywords:
osteoporosis;
Gufang Granules;
network pharmacology;
molecular dynamics simulations;
osteoclast
- From:
Chinese Journal of Information on Traditional Chinese Medicine
2025;32(3):42-50
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the potential targets and mechanism of Gufang Granules in treating osteoporosis through network pharmacology,molecular dynamics simulations,and in vitro experiment validation.Methods The active components of Gufang Granules were obtained from the TCMSP database and literature,and their related targets were predicted using SwissTargetPrediction database.Core drug targets were selected through protein-protein interaction(PPI)network analysis and machine learning models,and the predictive performance of the models was assessed by drawing receiver operating characteristic(ROC)curves on independent validation datasets.Gene Set Enrichment Analysis(GSEA)was used to analyze the expression and pathways of core targets.Molecular dynamics(MD)simulations were applied to evaluate the structural stability and interactions of the compound-target complexes.Non-cytotoxic concentrations of Gufang Granules containing serum were determined by the CCK-8 assay.RAW264.7 cells were treated with low,medium,and high concentrations of drug containing serum,respectively.The number of osteoclasts was quantified using TRAP staining.The expression levels of relevant genes and proteins were analyzed through qRT-PCR and Western blot methods.Results A total of 251 potential active components and 1 078 related targets of Gufang Granules were identified.The high expressions of core targets SRC and TNF were mainly associated with osteoclast differentiation,MAPK signaling pathway and PI3K/Akt signaling pathway.MD simulations showed that the core active component Glabridin exhibited strong stability and interaction with the SRC and TNF target proteins.The number of TRAP positive cells in all concentration groups of Gufang Granules was significantly reduced compared to the RANKL group(P<0.01,P<0.001).The serum containing Gufang Granules significantly reduced the mRNA expression of NFATc1,CTSK,SRC and TNF-α,and also downregulated the protein expression of NFATc1,CTSK,p-SRC and TNF-α(P<0.05,P<0.01,P<0.001).Conclusion Gufang Granules may inhibit osteoclast differentiation by downregulating the expression of NFATc1,CTSK,p-SRC and TNF-α,thereby slowing the pathological progression of osteoporosis.
