Network-based Pharmacological Analysis Of Core Herbs And Mechanisms Of Action For Improving Reduced Ovarian Reserve Function

OBJECTIVE: A preliminary investigation of the drug combination pattern of TCM for DOR treatment by means of data mining technology,a search for core drugs to improve DOR,and an in-depth analysis of the mechanism of action of core drugs for DOR treatment by means of network pharmacology.METHODS: The literature on the use of Chinese medicine to improve DOR in CNKI,Wan Fang,Sinomed and Pub Med databases was used as the base information,and prescriptions that met the nadir criteria were standardized and incorporated into Excel 2019 to obtain the Chinese medicine prescription datasets.The TCM Inheritance support system V3.0 was also used to conduct statistical and correlation rule analysis on the frequency of drug use,medicinal properties and flavours,the attribution of meridians and efficacy to explore the core drugs for improving DOR.The TCMSP and Gene Cards analysis platforms were then used to obtain the active ingredients and targets of the core drugs Shu Dihuang,Tu Sizi and Dang Gui as well as the relevant DOR disease targets.The two targets were extracted into the Venny2.1online tool to obtain the relevant intersection targets.Cytoscape 3.8.2 was used to visualise the intersecting targets and drug components,and to map the drug-chemical component-target network.The STRING platform was used to construct PPI networks and perform topological analysis to screen the important targets of Shu Dihuang,Tu Sizi and Dang Gui for the treatment of DOR.Finally,the Metascape database was used to perform GO and KEGG enrichment analysis on the relevant intersecting targets.RESULTS:1.A total of 231 articles covering 275 prescriptions and 213 herbal medicines were classified in this study,with a total frequency of 3347.Drug use was most common in the category of tonifying deficiency(59%)and invigorating blood circulation and resolving blood stasis(12%).The properties of the medicines were mainly warm(42%)and flat(30%),the taste of the medicines was mainly sweet(67%),bitter(40%)and pungent(29%),and the meridians of the medicines were mainly liver(29%)and kidney(23%).The commonly used drugs for improving DOR were Shu Dihuang,Tu Sizi,Dang Gui,Shan Yao,Shan Zhuyu,Gou Qizi and Bai Shao.After association rule analysis,74 drug combinations with high association for treating DOR were obtained,among which Tu Sizi-Dang Gui,Shu Dihuang-Dang Gui and Shu Dihuang-Tu Sizi were the core drug combinations.2.With the help of the TCMSP platform,13 effective chemical components were finally screened,involving Beta-sitosterol,Quercetin,Stigmasterol and sesamin,etc,while 219 corresponding targets were obtained.The results of PPI topology analysis showed that the active ingredients of Tu Sizi and Dang Gui played key roles in the treatment of DOR,and quercetin,kaempferol and beta-sitosterol were the main ingredients for the treatment of DOR,while JUN,TP53,AKT1,TNF,MAP,etc.were the main ingredients for the treatment of DOR,The results of GO functional enrichment analysis showed that Shu Dihuang,Tu Sizi and Dang Gui were able to improve ovarian reserve and treat DOR by responding to hormonal and exogenous stimuli,reducing cell population proliferation and regulating apoptotic signaling pathways,The results of KEGG pathway enrichment analysis showed that Shu Dihuang,Tu Sizi and Dang Gui were able to treat DOR by regulating Cancer,AGE-RAGE and PI3K/Akt signaling pathways.CONCLUSION:1.In the treatment of DOR,Chinese medicine selects medicines that are warm and flat,with sweet,bitter and pungent properties,mainly belonging to the liver and kidney meridians,and mostly using medicines that nourish the liver and kidney and invigorate blood circulation to remove blood stasis.Therefore,tonifying liver-kidney and promoting blood circulation therapy is the basic treatment for this disease.The use of Shu Dihuang,Tu Sizi and Dang Gui in combination with each other can provide reference for clinical optimization of prescription.2.The active ingredients in Shu Dihuang,Tu Sizi and Dang Gui can improve Ovarian reserve and exert therapeutic effects by regulating several target genes and participating in the regulation of corresponding biological processes and signalling pathways.3.Quercetin,beta-sitosterol and kaempferol are the core components of the drug,which can act on JUN,TP53,AKT1,TNF,MAPK1,IL6 and other key target genes,participating in the response to hormonal and exogenous stimuli,reducing cell population proliferation,regulating apoptotic signalling pathways and other links to play a role.4.Cancer,AGE-RAGE,PI3K/AKT and other signalling pathways are the pathways significantly associated with DOR,and such pathways mainly interfere with follicular growth and development.It is suggested that Shu Dihuang,Tu Sizi and Dang Gui focus on the treatment of DOR by positively regulating the development of follicles and improving Ovarian function.