| Objective To explore the effect and mechanism of Tiaojing decoction in the treatment of premature ovarian insufficiency through animal experiment and network pharmacology.Methods(1)Animal experiment:Thirty-two 12 weeks old female SD rats without mating and with regular estrous cycle were randomly divided into four groups:blank group,model group,western medicine group and traditional Chinese medicine group.The rats in the blank group were intraperitoneally injected with sterile water for injection at the body weight of 0.4 mL/100 g,and the rats in the other groups were intraperitoneally injected with cisplatin solution at the concentration of 0.5 mg/mL for 7 consecutive days to establish the rat model of immune premature ovarian insufficiency.After modeling,the blank group and model group were given equal volume of normal saline by gavage;The western medicine group was given estradiol valerate(trade name:Bujiale)solution with a concentration of 1.25 mg/mL,0.12 mg/(kg·d);The traditional Chinese medicine group was given 1 g/100 g body weight Tiaojing decoction by gavage,twice a day,for 14 consecutive days.The serum levels of follicle stimulating hormone(FSH),anti-Müllerian hormone(AMH)were detected by Elisa.HE staining were used to observe the morphological changes of one side of rat ovary.(2)Network pharmacology:Kyoto encyclopedia of genes and genomes(KEGG)DRUG database,traditional Chinese medicine database and analysis platform(TCMSP)were used to mine the molecular structure of estradiol valerate and the potential active components and drug targets of Tiaojing decoction.POI related targets were screened through the human gene database(GeneCards),online mendelian inheritance in man(OMIM)and pharmacogenomics knowledgebase(PharmGKB)databases.The intersection target obtained by matching the drug targets with the disease targets were used to establish the traditional Chinese medicine component targets disease action network by using cytoscape 3.8.0 software.The protein interaction of the obtained intersection targets was analyzed by string website,and the enrichment analysis of gene ontology(GO)and KEGG was carried out by R software.Results(1)Animal experiments:1.Appearance changes:Compared with the model group,the rats in the western medicine group and the traditional Chinese medicine group gradually recovered their mental state with the increase of administration time,increased their weight rapidly and acted quickly,the recovery of traditional Chinese medicine group was better than that of western medicine group;2.Weight change:After 14 days of administration,the weight of rats in four groups increased in varying degrees,except that there was significant difference between the model group and the blank group(P<0.05),there was no significant difference between the western medicine group,the traditional Chinese medicine group and model group(P>0.05),and there was no significant difference between the two groups(P>0.05);3.Estrous cycle change:the estrous cycle of the blank group showed regular changes,and the estrous cycle of the other three groups lost normal regular changes after modeling,mainly manifested in the extension of estrous interval and pre estrous period,and the shortening or disappearance of estrous period.After administration for a period of time,the estrous cycle of rats in western medicine group and traditional Chinese medicine group gradually returned to regular changes,and the recovery of estrous cycle in model group was the worst;4.Morphological changes of ovarian tissue:the morphological structure of ovarian tissue in the blank group was clear,and follicles of different grades were visible.The endometrium was thick.The ovarian tissue in the model group atrophied and the endometrium was thin.The morphological changes of ovarian tissue in the western medicine group were significantly improved compared with those in the model group,and the endometrium was thickened.The morphological recovery of ovarian tissue in the Chinese medicine group was the best,the endometrium was significantly thickened,and the number of mature follicles was the most;5.Comparison of serum hormones:Compared with the blank group,there were no significant difference in serum FSH and AMH between the other three groups(P>0.05).Compared with the model group,there were no significant difference in serum FSH and AMH between the other three groups(P>0.05),and there was no statistical significance between the western medicine group and the traditional Chinese medicine group(P>0.05).(2)Network pharmacology:A total of 181 active components,2 873 potential targets and 4 597 POI targets were screened from Tiaojing decoction.192 key genes were screened by comparing the common targets of Tiaojing decoction and POI.The main active ingredients are quercetin,kaempferol,luteolin,β-sitosterol and stigmasterol,the core gene is hypoxia inducible factor 1 subunit alpha(HIF1A),mitogen activated protein kinase 1(MAPK1),JUN proto-oncogene(JUN),estrogen receptor 1(ESR1),MYC proto-oncogene(MYC),tumor protein p53(TP53),signal transducer and activator of transcription 3(STAT3).The biological processes are mainly the reaction of oxides and oxidative stress,the response of cells to chemical stress and material metabolism,which plays molecular functions such as transcription and binding.The biological pathways include interleukin-17(IL-17)signaling pathway,tumor necrosis factor(TNF)signaling pathway and phosphatidylinositol 3 kinase(PI3K)-protein kinase B(AKT)signaling pathway,mitogen-activated protein kinase(MAPK)signaling pathway,forkhead box transcription factor O(FoxO)signaling pathway,estrogen signaling pathway,Wnt signaling pathway,etc.The target protein of estradiol valerate was estrogen receptor,and KEGG pathway was estrogen signaling pathway.Molecular docking results showed that quercetin had the best binding degree with HIF1 A,and the binding value was 6.78.Conclusion(1)Tiaojing decoction can restore the morphological structure of ovary,thicken endometrium and improve ovarian reserve function in cisplatin induced POI rats;(2)Tiaojing decoction mainly plays a role in the treatment of POI through potential biological processes such as response to oxide and oxidative stress,response to chemical stress stimulation and material metabolism response,and with the help of multiple potential pathways such as immunity,signal transduction and hormones;(3)Compared with estradiol valerate,Tiaojing decoction through multiple components,targets and pathways to treating POI. |
