多囊卵巢综合征模型小鼠卵巢颗粒细胞内关键差异表达基因的挖掘OA

In order to identify key differentially expressed genes(DEGs)in ovarian granulosa cells(GCs)of mice with polycystic ovary syndrome(PCOS)model and to preliminarily investigate their regulatory roles in PCOS pathogenesis,36 three-week-old female ICR mice were randomly divided into the control and dehydroepiandrosterone(DHEA)groups,with 18 mice in each group.The control group was subcutaneously injected with 0.1 mL of a sesame oil and ethanol mixture(10%ethanol and 90%sesame oil)daily,while the DHEA group was subcutaneously injected with 0.1 mL of a DHEA suspension(DHEA[6 mg/100 g]dissolved in a 10%ethanol and 90%sesame oil solution)daily for 20 consecutive days.After modeling,both groups of mice were assessed for estrous cycle,morphological changes,and serum hormone levels.The GCs were randomly collected from three mice for transcriptome sequencing(RNA-Seq),and DEGs were identified and verified by RT-qPCR.The results showed that in the DHEA-treated group,the estrous cycle of ICR mice was disrupted,with prolonged estrus and a reduced duration of estrous periods,and the absence of proestrus and metestrus,indicating an incomplete and irregular estrous cycle.The number of cystic follicles increased,and corpora lutea were not observed,resulting in a polycystic ovary morphology.The mass concentration of testosterone,progesterone,the activity of luteinizing hormone,and the ratio of luteinizing hormone to follicle-stimulating hormone were significantly higher in the DHEA group compared to the control group(P<0.05 or P<0.01),while the activity of follicle-stimulating hormone was significantly lower(P<0.05).A total of 161 DEGs were identified.Four DEGs(ICAM1,MRO,NR4A1,and VCAM1)that had been confirmed to be related to PCOS were selected for RT-qPCR validation.The FPKM value of ICAM1,VCAM1,MRO,and NR4A1 were significantly higher in the DHEA group compared to the control group(P<0.01),and the expression trends were consistent with the RNA-Seq,proving the reliability of the sequencing results.Subsequently,five candidate genes,COMP,CMTM1,CAR4,IRF7,and MXRA8,which had not been reported in the literature,were selected for verification.The relative mRNA expression levels of COMP,CMTM1,IRF7,and MXRA8 in the GCs were significantly higher in the DHEA group compared to the control group(P<0.01),while the relative mRNA expression of CAR4 was significantly lower(P<0.01),and the expression trends were consistent with the RNA-Seq.In conclusion,this study had successfully constructed PCOS mouse model;five new DEGs with potential research value were identified.These genes might be involved in the regulation of PCOS pathogenesis by mediating inflammatory responses,ferroptosis,or insulin resistance,providing new targets and directions for future studies on the molecular mechanisms of PCOS.