Investigation Of The Mechanisms Of Quanduzhong Capsule In Osteoporosis Based On The Integration Strategy Of Network Pharmacology And Metabolomics

Osteoporosis(OP)is a metabolic disease with disturbances in bone formation and bone resorption,which usually occurs in the elderly.The number of people suffering from osteoporosis will rise progressively as the population aging becoming a severe problem in the future,and the stress of medical and pharmaceutical care will rise also.Quanduzhong Capsule(QDZC)is a modern traditional Chinese medicine preparation composed of the cortex powder of Eucommia ulmoides Oliv.(Duzhong)and its extract,which is recorded on the Pharmacopoeia of the People’s Republic of China 2015 and 2020 Edition.It plays a role in reducing blood pressure,nourishing the liver and kidney,and reinforcing the muscles and bones and applies to anti-hypertension and anti-osteoporosis clinically.But the mechanism of QDZC on OP is not yet clear.The study was focused on QDZC and integrated the techniche of network pharmacology and metabolomics to investigate the mechanisms of QDZC in osteoporosis based on the identification of effective components of QDZC comprehensively.1.Identification of chemical components of QDZC86 components were identified by Ultra performance liquid chromatography combined with quadrupole time-of-flight mass through(UPLC-Q-TOF-MS)comparing the retention time,UV absorption,exact molecular weight,MS fragment,reference standards and references.There are 31 lignans,13 iridoids,12 organic acid,7 phenylpropanoids,2 aldehydes,1 alcohol,1 coumarin,1 sesquiterpene,1 sugar,1 triterpene,9 other compounds and 7 unknow compounds.2.Network pharmacology of compounds in QDZC1374 targets related with osteoporosis were found in diseases database such as Drugbank,TTD,etc.334 targets related with compounds in QDZC were found in some Chinese medicine such as ETCM,HERB,etc.STRING database and Cytoscape software were applied to create Protein-protein interaction.9 compounds named Eucommiol II,Asiatic acid,(+)1-hydroxypinoresinol,isochlorogenic acid A,aucubin,neochlorogenic acid,pinoresinol,catalpol,and chlorogenic acid and 19 core targets such as MYB,TNFRSF1 A,NOS2,SMAD3,SMAD2 were filtered by Cyto CNA.14 core pathways which degree value was greater than10 were screen out from 143 pathways which were enriched by DAVID database,including lipid and atherosclerosis,PI3K-AKT signaling pathway,endocrine resistance,MAPK signaling pathway,and Fox O signaling pathway.When biding the compounds’ targets and pathway together on one map,we found that most compounds target on estrogen,PI3K-AKT,MAPK,Fox O,TNF,Wnt,IL-17 signaling pathway.3.Metabolomics study of OP of QDZCThe study of metabolomics was established on the ovariectomy osteoporosis rat model and investigated the index of body weight(BW),bone mineral density(BMD),and the expression of AKT,p-AKT,ERK,p-ERK,and ER in serum of Sham,Model,Model+QDZC(1000mg/kg/day),Model+Alendronate Sodium Tablet(6 mg/kg/week).And the difference of serum metabolites in each group was analyzed by UPLC-Q-TOF-MS at 4,8 and 12 weeks.The BW of group QDZC at week 8 and 12 was significantly lower than the group model while the BMD of group QDZC at week 8 was lower than group model but not significantly.The week 4 and 8 result of Elisa test on AKT,p-AKT,ERK,p-ERK indicated that QDZC may regulate and control the PI3K-AKT and MAPK signaling pathway to play anti-osteoporosis roles.Through metabolomics of serum,25 metabolites were filtered out which related with the arachidonic acid,biosynthesis of unsaturated fatty acid,steroid biosynthesis,primary bile acid biosynthesis,and glycerophospholipid metabolism.Combined with literatures,the mechanism of QDZC for anti-osteoporosis may be associated with inhibiting inflammation,osteoclast generation and bone resorption,activating the PI3K-AKT and ERK signaling pathway,increasing the synthesis of calcitriol to raise calcium absorption.In conclusion,the study combines UPLC-Q-TOF-MS to identify compounds in QDZC,network pharmacology to analyze the core effective components and targets and metabolism to analyze the metabolites which QDZC may affect.It tentatively explored the antiosteoporosis mechanism of QDZC and provided basis for the clinical application of QDZC.