Preliminary Study On The Molecular Mechanism Of Morinda Officinalis On Polycystic Ovarian Syndrome

Objective: Through the effective integration of multiple datasets in bioinformatics,the signal pathways and efficient target molecules related to polycystic ovary syndrome(PCOS)were deeply explored.Animal experiments were conducted to preliminarily explore the impact of Morinda officinalis on the occurrence and development of PCOS.Furthermore,network pharmacology was used to analyze the gene targets and signal pathways of Morinda officinalis on PCOS,in order to discover the potential mechanism and key target molecules of Morinda officinalis on PCOS,Provide new ideas for the research of Haiba Morinda in the treatment of PCOS.Provide new targets and pathways for the early diagnosis and clinical treatment of PCOS,and also provide more effective methods for the clinical diagnosis and treatment of PCOS.Method:1.Screening key target molecules for polycystic ovary syndrome based on bioinformatics and preliminary validation(1)Using the GEO2 R platform to analyze PCOS microarray related data obtained from the NCBI database and m RNAs microarray screening of GSE106724(8 patients and 4 normal individuals)data,P<0.05 was compared with screening differentially expressed genes in PCOS patients using 1 as the standard(2)Obtain the PPI network diagram and related data of differentially expressed gene cross targets through the STRING database,and construct the network diagram using Cytoscape 3.9.0(3)Perform gene functional analysis on differentially expressed genes through R4.0.5 platform,provide KEGG analysis,and use pathview to draw a signal pathway map with the maximum GeneRatio value.(4)Use RT qPCR technology to verify the relative m RNA expression levels of predicted key genes in follicular fluid samples from patients with polycystic ovary syndrome and normal human follicular fluid samples.2.Based on network pharmacology,preliminary exploration of the molecular mechanism of Morinda officinalis in the treatment of polycystic ovary syndrome(1)Animal experiment: SD female rats were randomly divided into three groups:normal control group(n=5),PCOS model group(n=5),and Morinda officinalis gavage group(n=5).HE staining and electron microscopy were used to observe the histological morphology of the ovaries.(2)Retrieve the effective ingredients of Morinda officinalis through TCMSP,TCMID,NPASS,and BATman traditional Chinese medicine platforms,and then set screening conditions based on the pharmacokinetic parameters of the ingredients.Select active ingredients that meet the drug value(DL)≥ 0.18 and oral efficacy(OB)≥ 30%,and obtain chemical components with certain biological activity in Morinda officinalis.(3)Potential gene targets for chemical components identified through TCMSP,NPASS,Swiss Target Prediction,and TCM-ID platforms.(4)Gene targets significantly associated with polycystic ovary syndrome were identified through the Dis Ge NET,OMIM,and DRUGBANK platforms.(5)By comparing the potential gene targets of drug action retrieved from different platforms(2)and(3)and their correlation with polycystic ovary syndrome,common gene targets were selected and Venny2.1 was used to draw Venn map.(6)Import the selected common gene targets into the STRING database platform,use Cytoscape software for visual analysis,and establish a protein protein interaction network(PPI)from all directions,multiple angles,and deep levels to screen key gene targets.(7)Through DAVID online database,the selected key gene targets are annotated with Gene Ontology(GO)functions to clarify the biological attributes of key gene targets.(8)The main pathways for identifying key gene targets were identified through cluster induction analysis using the Kyoto Encyclopedia of Genes and Genomes(KEGG).Result:1.Screening key target molecules for polycystic ovary syndromeBioinformatics analysis found that in cases of polycystic ovary syndrome,the expression levels of differentially expressed genes extracted from the GEO database were upregulated by 310 genes and downregulated by 297 genes.The PPI network analysis results indicate that ALPL,SPP1,FKBP5,and AKAP5 are key target molecules for polycystic ovary syndrome.GO functional enrichment analysis indicates that these genes are involved in cell response to interferon γ Such as chemokine receptor binding,MHC class II protein complexes,and endoplasmic reticulum membrane components.KEGG enrichment analysis showed that these genes were mainly involved in asthma,rheumatoid arthritis,allograft rejection,type I diabetes and other signaling pathways.The RT-q PCR experiment results showed that ALPL was low expressed in patients with polycystic ovary syndrome,while SPP1,FKBP5,and AKAP5 were high expressed.2.The potential molecular mechanism of the effect of Morinda officinalis on polycystic ovary syndromeMorphological changes in the ovaries of rats: In the normal control group,follicles at all levels of development can be seen in the ovaries,with mostly 8-9 layers of granulosa cells and a structurally normal corpus luteum;In the PCOS model group,the number of granulosa cell layers in the ovaries of rats decreased,with cystic expansion of follicles and an increase in atresia follicles.No oocytes or corona radiata were observed;The proportion of follicles at all levels in Morinda officinalis gavage group was normal,and the number of granulosa cell layers increased.Network pharmacology analysis found that Morinda officinalis contains 17 components,including daidzein,p-hydroxycinnamic acid and hispidulin,which have good ADME properties.17 effective active ingredients can target 10 gene targets related to polycystic ovary syndrome.After constructing the PPI network diagram,seven key gene targets were selected,namely PIK3 CD,MTOR,CDC42,SLC9A1,ODC1,EIF2AK3,and CXCR4.These seven key gene targets are involved in a variety of signal pathways,including the regulation of actin cytoskeleton,migration of leukocytes through endothelial cell migration,thyroid hormone,Erb B,chemokines,VEGF,etc.Conclusion: Bioinformatics analysis indicates that effective target molecules ALPL,SPP1,FKBP5,and S are involved in the occurrence and development of polycystic ovary syndrome.The RT q PCR experiment also confirmed this result,indicating that differentially expressed genes in polycystic ovary syndrome are involved in biological processes and signaling pathways related to immune and inflammatory responses.Animal experiments have preliminarily verified the impact of Morinda officinalis on the occurrence and development of PCOS.Network pharmacology analysis shows that Morinda officinalis plays a role in polycystic ovary syndrome through gene targets PIK3 CD,MTOR,CDC42,SLC9A1,ODC1,EIF2AK3,and CXCR4.The screened active ingredients,gene targets,and signaling pathways are all related to inflammatory response.Therefore,Haiba Morinda officinalis may mainly have an anti-inflammatory effect on polycystic ovary syndrome.