Study On Anti-menopausal Osteoporosis Of Mongolian Prescription Tubson-2 Decoction Based On Network Pharmacology

Objective The Mongolian medicine decoction Tubson-2(Tubson-2 decoction,TBD)is often used in Mongolian medicine clinics for bone healing.Some scholars have used the Mongolian medicine Tubson-2 decoction to treat post-menopausal osteoporosis(Post-menopausal osteoporosis,PMOP)found that the curative effect is obviously,but the the potential substance basis and mechanism of action are still unclear.In this study,the network pharmacological analysis method is used to predict the key target of the Mongolian medicine Tubson-2 in the treatment of post-menopausal osteoporosis.At the same time,animal experiments are used to verify its anti-postmenopausal osteoporosis efficacy and mechanism of action,and the Mongolian medicine tower is preliminarily clarified.The mechanism of action of Tubson-2 against post-menopausal osteoporosis.In addition,from the perspective of metabonomics,the serum,urine,and fecal samples of the pharmacodynamic experiment part are analyzed,and the Mongolian medicine Tubson-2 treats post-menopausal osteoporosis caused by metabolite changes and its effect on metabolic pathways.The regulation function provides scientific basis for the clinical application of the Mongolian medicine Tubson-2.Method 1.Use multiple databases to enrich all the chemical components of Echinops latifolius Tausch and Eucommia ulmoides Oliv.in the Tubson-2,and establish it through Swiss and Superpred databases The compound target library of Tubson-2.Search the Drugbank database to establish a disease target library for post-menopausal osteoporosis,compare the compound target library with the disease target library,find a common target,use Cytoscape to perform topological analysis,and establish a corresponding association network model.KEGG pathway analysis was performed on the common targets through the DAVID database to predict the key target of Tubson-2 decoction against post-menopausal osteoporosis.2.Establish a model of osteoporosis in ovariectomized rats,set up a blank group,a model group,a sham operation group,and treated groups.The treated groups include the low,medium and high doses of the Tubson-2,the Gushukang group and the estrogen group.8weeks for administered,after the last administration,they entered the metabolic cage and fasted for 24 hours and collected urine and feces.Serum and left tibia samples were collected.X-ray was used to detect bone microstructure;ELISA was used to determine the expression of HSD11B1,CYP19A1,and VDR,the targets of anti-menopausal osteoporosis.3.Use UPLC-Q-Exactive MS to perform non-targeted metabolomics analysis on rat serum,urine,and fecal,and perform multivariate statistical analysis on the data through Simcap software.Using PCA,PLS-DA,OPLS-DA and other pattern recognition methods to screen out the VIP value>1 and P<0.05 differential metabolites as potential characteristic biomarkers.Then the potential biomarkers are enriched in pathways and their biological significance is analyzed.4.Use molecular docking technology to dock the compound directly connected to the key target to obtain the compound with the lowest binding energy,and analyze it.Results 1.A total of195 compounds were enriched,of which 62 active ingredients obtained 2251 targets,and Tubson-2 decoction had 581 disease-related targets.After integration,a total of 89 targets were obtained.According to topological analysis,the key targets of the Tubson-2 decoction for the treatment of post-menopausal osteoporosis are: VDR,CYP19A1,HSD11B1.2.The Tubson-2 extract can be used to treat post-menopausal osteoporosis.Increase the bone density,bone volume/total volume,volume fraction and number of bone trabeculae in ovariectomized rats,reduce the separation of bone trabeculae,and improve the bone microstructure of ovariectomized rats.The Mongolian medicine Tubson-2 decoction plays a role in treating post-menopausal osteoporosis by down-regulating the expression of VDR and CYP19A1 in the serum.3.After the treatment of the Mongolian medicine Tubson-2 decoction,through the analysis of serum metabolomics,we found that the Tubson-2 mainly regulates the metabolism of Vitamin B6;through the urine metabolomics analysis,we found that the Tubson-2 mainly regulate histidine metabolism;through fecal metabolomics analysis,we found that the Tubson-2 decoction mainly regulates the metabolism of Vitamin B6 to treat post-menopausal osteoporosis.4.There are 5 compounds in the Tubson-2 that are directly connected to the key target VDR,then docked them with VDR in turn.The results show that Helenalin,a component from Eucommia ulmoides Olive,has the lowest binding energy to the VDR.Conclusion: In this study,taking the Mongolian medicine Tubson-2 decoction as an example,with the molecular mechanism as the main line,firstly verify the efficacy of the Mongolian medicine Tubson-2 against post-menopausal osteoporosis,and then integrate network pharmacology and metabolomics,network pharmacology prediction started and the metabolism ended,and the key target VDR was found.Finally,molecular docking technology was used to find the potential active ingredient Helenalin in the Mongolian medicine Tubson-2,indicating that the Mongolian medicine Tubson-2 mainly regulates the content of VDR in the serum through Helenalin,thereby interfering with vitamin B6 metabolism disorders,and then treating post-menopausal osteoporosis.Above researchs enrich the molecular mechanism of the Mongolian medicine Tubson-2 against post-menopausal osteoporosis: a holistic view of vitamin B6 metabolism.Our research provides new ideas for the mechanism study,and provides a direction for the follow-up experimental research of the Mongolian medicine Tubson-2.It also has a certain guiding effect on the follow-up development of this prescription,and provides a new basis for guiding the clinical rational use of drugs.