Screening And Primary Validation Of Key Genes And Pathways In Age-Associated Ovarian Aging Based On GEO Database

【Objective】Ovarian aging leads to reproductive and endocrine dysfunction,causing the disorder of multiple organs and even declined quality of offspring’s health.Age is the main factor that causes ovarian aging.However,few studies have investigated the changes in gene expression profile in the age-associated ovarian aging process.Here,we applied integrated bioinformatics to screen,identify,and validate the critical pathogenic genes involved in age-associated ovarian aging and uncover potential molecular mechanisms and potential biomarkers,providing potential targets for treating age-associated ovarian aging.【Methods】The data of age-associated ovarian aging was screened and the expression profiles of GSE84078 were downloaded from the Gene Expression Omnibus(GEO)database to compare the mRNA expression levels of ovaries from young and old mice.The expression profiles of GSE84078 included the data from ovarian samples of 10 normal C57BL/6 mice,including old(21–22 months old,ovarian failure period)and young(5–6 months old,reproductive bloom period)ovaries.Differentially expressed genes(DEGs)were filtered.Functional enrichment analysis was performed.The hub genes were identified by constructing a protein–protein interaction(PPI)network and consequently verified using qPCR and immunohistochemistry.The immune cell infiltration in the age-associated ovarian aging process was evaluated by applying CIBERSORT,and a correlation analysis between hub genes and immune cell type was also performed.【Results】First,we filtered 931 DEGs,including 876 upregulated and 55 downregulated genes.Functional enrichment analysis showed that biological functions of DEGs were primarily immune response regulation,cell–cell adhesion,and phagosome pathway.The most closely related genes among DEGs were Tyrobp,Rac2,Cd14,Zap70,Lcp2,Itgb2,H2-Ab1,and Fcer1g.Tyrobp,Rac2,Cd14,Zap70,Lcp2,Itgb2,H2-Ab1 and Fcer1g were verified by qPCR;Lcp2,H2-Ab1,Itgb2 and Cd14 were verified by immunohistochemistry.The results of the immune cell infiltration and the correlation analysis between hub genes and immune cell type suggested that plasma cells and na(?)ve CD4~+T cells may participate in age-associated ovarian aging.The hub genes were positively correlated with memory B cells,plasma cells,M1 macrophages,Th17 cells,and immature dendritic cells.【Conclusions】In conclusion,this study indicates that hub genes such as Tyrobp,Rac2,Cd14,Zap70,Lcp2,Itgb2,H2-Ab1 and Fcer1g participate in the process of age-associated ovarian aging by regulating the immune system.Screening for DEGs and pathways in age-associated ovarian aging using bioinformatic analysis could provide potential clues for researchers to unveil the molecular mechanism underlying age-associated ovarian aging and provide effective molecular targets for the treatment of age-associated ovarian aging.