Analysis of mechanism of Shenwei Gubi Tang(参威骨痹汤)against osteoporosis based on the network pharma-cology approach,molecular docking techniques and animal experimentation
Objective:To explore the mechanism of Shenwei Gubi Tang(参威痹汤,SWGBT)against osteoporosis(OP).Methods:①Network pharmacology research.The ingredients in SWGBT were identified by using ultra-high performance liquid chromatography-qua-drupole-orbitrap mass spectrometry(UHPLC-QE-MS),and the key active ingredients and action targets of SWGBT against OP were prelimi-narily screened using the network pharmacology approach.②Validation by molecular docking.The binding effects between the key active in-gredients and the targets of SWGBT against OP determined by network pharmacology were validated by molecular docking techniques,and the combination with the best binding effect was screened.③Verification by animal experimentation.Sixty female C57BL/6JNifdc mice were selected and randomized into model group,low-dose SWGBT(L-SWGBT)group,medium-dose SWGBT(M-SWGBT)group,high-dose SWG-BT(H-SWGBT)group and sham-operated group.The mice in model group,L-SWGBT group,M-SWGBT group and H-SWGBT group were subjected to bilateral ovariectomy for inducing OP;while the ones in sham-operated group were merely removed an equal volume of peri-ovarian adipose tissues.After successful modeling,the mice in L-SWGBT group,M-SWGBT group and H-SWGBT group were intervened by intragastric administration with 14.95,29.9,59.8 g/(kg·d)SWGBT,respectively;while the ones in sham-operated group and model group with the same dose of normal saline.After 8-week drug intervention,the rats were sacrificed and their femurs were harvested for Micro-CT examination and observation on histopathological changes through HE staining and tartrate-resistant acid phosphatase(TRAP)staining.Fur-thermore,the mRNA expression levels of key target genes(determined by network pharmacology approach and molecular docking tech-niques)in tibia tissues were detected by using real-time quantitative PCR(RT-qPCR).Results:①The results of network pharmacology re-search showed that SRC,mitogen-activated protein kinase 3(MAPK3),MAPK1,phosphatidylinositol-3-kinase regulatory subunit 1(PI3KR1),and signal transducer and activator of transcription 3(STAT3)were the key targets of SWGBT against OP,and the schisandrin C,10-gingerol,butyl p-hydroxybenzoate,sesquiterpene lactone,and coumestrol were the key active ingredients of SWGBT against OP.②The results of molecular docking showed that the key active ingredients of SWGBT against OP had good binding ability to the key targets with the binding energy less than-5.0 kJ/mol,and the targets including SRC,MAPK3,and PI3KR1 exhibited better binding effects with the active ingredients including schisandrin C,10-gingerol,and butyl p-hydroxybenzoate.③The results of Micro-CT examination and histopathological observation showed that the SWGBT could improve the degree of OP in OP model mice,with the medium-dose SWGBT displaying better effects.The results of RT-qPCR showed that the mRNA levels of SRC and MAPK3 were lowly expressed in the tibia tissues of OP model mice in M-SWGBT group compared to model group,while,the mRNA expression level of P13KR1 was not significantly different from that of model group.Conclusion:The SWGBT may regulate bone metabolism for treatment of OP through adjusting the expression of SRC,PI3KR1 and MAPK3 via the ingredients such as schisandrin C,10-gingerol and butyl p-hydroxybenzoate.
osteoporosisShenwei Gubi Tangnetwork pharmacologymolecular docking simulationanimal experimentation
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