Research On Zishen Huoxue Tiaojing Decoction And Femoston For POI With Kidney Deficiency And Blood Stasis Based On Clinical Investigation And Network Pharmacology

Objective:The aim of this paper is to examine the clinical efficacy of a modality combining a self-formulated traditional Chinese medicine,zishen huoxue tiaojing decoction,and Femoston for the treatment of premature ovarian insufficiency（POI）in patients with concurrent kidney deficiency and blood stasis.Furthermore,network pharmacology is also utilized to analyze the mechanism of the proposed recipe in the treatment of POI.In conclusion,it is shown that the integration of traditional Chinese and Western medicine is used to treat POI with kidney deficiency and blood stasis in this letter,which provides a preliminary clinical exploration and theoretical verification of the efficacy and safety of the combination therapy.Methods:1.From January 2022 through December 2022,we enrolled 64 POI patients who met the study criteria and are diagnosed with kidney deficiency and blood stasis type,from the gynecology outpatient department at the First Affiliated Hospital,Heilongjiang University of Chinese Medicine.Following a randomized controlled design,the patients were allocated into an experimental group（including 32 patients）and a control group（including 32patients）.The experimental group received the self-formulated traditional Chinese medicine,zishen huoxue tiaojing decoction,in combination with Femoston,while the control group was treated with Femoston alone for three menstrual cycles.It was observed and compared for changes in menstrual cycle,traditional Chinese medicine diagnostic scores,serum hormone levels（FSH,LH,E₂）,serum AMH levels,depression score,anxiety score,and Kupperman score,before and after treatment.Then,the corresponding clinical treatment data was collected,organized,analyzed,and finally,the conclusions were obtained.2.Utilize a network pharmacology approach to investigate the mechanism of zishen huoxue tiaojing prescription in treating POI.The TCMSP database was used to retrieve the active ingredients and target genes of the 10 herbs in the proposed formula,which were then imported into the Uni Prot database and standardized.The corresponding disease genes associated with POI were obtained in the Genecards,OMIM,PHARMGKB,and Dis Genet disease databases.Then,A Venn diagram was generated by intersecting the drug and disease genes,and a"drug-active ingredient-target"network was constructed by importing the overlapping genes into Cytoscape software.With the aid of the String database,the PPI network of the overlapping targets was given,and critical targets were identified via R language and Cytoscape software.Finally,GO and KEGG enrichment analysis of the related targets was performed by using the Metascape platform.Results:Clinical Observation:1.There existed no statistically significant differences（P>0.05）in age,disease duration,quantified menstrual condition scores,TCM diagnostic scores,serum FSH,LH,E₂and AMH levels,depression score,anxiety score,and Kupperman score between the two groups before treatment,indicating that the data were comparable.2.Following three menstrual cycles of treatment,both groups showed significant improvements in quantified menstrual condition scores,TCM diagnostic scores,serum FSH,LH,E₂levels,depression score,anxiety score,and Kupperman score（P<0.01）,while there was no significant improvement in serum AMH levels（P>0.05）.3.Following three menstrual cycles of treatment,for the experimental group,it was demonstrated that there was statistically significant superior therapeutic effects compared with the control group in terms of reducing quantified menstrual condition scores,TCM diagnostic scores,depression score,anxiety score,and Kupperman score（P<0.05）,as well as improving serum E₂levels and reducing serum FSH levels（P<0.05）.However,there was no statistically significant difference in improving serum LH levels and increasing serum AMH levels between the two groups（P>0.05）.4.According to the comprehensive efficacy,it was shown that the effective rate in the experimental group was 90.00%,while the counterpart in the control group was 73.33%,which was lower compared with the experimental group（P<0.01）.Network Pharmacology:After screening and standardization of 10 herbs in the self-formulated zishen huoxue tiaojing decoction in the TCMSP database,a total of 121 active ingredients and 127 related targets were obtained.Meanwhile,4309 potential disease targets were obtained from the database,among which 103 associated targets were identified by intersecting disease targets with drug targets.Then,the highly related active ingredients were analyzed and screened,including quercetin,kaempferol,jasminoidin,baicalin,β-sitosterol,stigmasterol,and so on.The main related genes were identified through PPI network analysis,which included ALB,ESR1,IL6,MYC,CTNNB1,EGFR,CASP3,etc.GO functional enrichment analysis showed that the biological processes mainly involved the response to hormones and the response of cells to nitrogen compounds and organic nitrogen compounds,etc,while the cellular components mainly related to the transcription initiation complex,lipid raft,membrane microdomain,and so forth.The molecular functions mainly involved transcription factor binding,DNA binding transcription factor binding,RNA polymerase II specific DNA binding transcription factor binding,and so on.KEGG pathway enrichment analysis exhibited that the target genes were mainly enriched in cancer pathways,chemical carcinogenesis-receptor activation,lipid and atherosclerosis,human cytomegalovirus infection,and the PI3K-Akt signaling pathway,AGE-RAGE signaling pathway in diabetic complications,and TNF signaling pathway.Conclusions:1.The combination therapy of the self-formulated zishen huoxue tiaojing decoction and Femoston was superior to using Femoston alone in improving menstrual condition,TCM diagnostic scores,depression scores,anxiety scores,and Kupperman scores.The efficacy was precise,safe,and without notable toxic side effects,which is worth promoting in clinical treatment.2.The self-formulated zishen huoxue tiaojing prescription combined with Femoston treatment showed significantly more efficacy than the control group in reducing serum FSH levels and increasing E₂levels.Both of them demonstrated equivalent efficacy in reducing serum LH levels,but had no significant effect on serum AMH levels.3.Based on network pharmacology analysis,the self-formulated zishen huoxue tiaojing recipe may treat POI through multi-component,multi-target,and multi-pathway modes,including anti-apoptosis,restoration of ovarian endocrine function,increase of blood supply,inhibition of inflammatory response,regulation of immune balance,and anti-oxidative stress.