Expression Of Sohlh1 And MiRNA-342-3p In Ovaries Of Perimenopausal Women

Objective:Perimenopause is a period of transition to menopause,a destructive process that can last for decades.Ovarian function is increasingly failing in perimenopausal women.Tissue-specific basic helix-loop-helix(bHLH)transcription factor proteins often play important roles in cell differentiation.The bHLH protein Sohlh1(Spermatogenesis and oogenesis specific basichelix-loop-helix transcription factor 1)is specifically expressed in oocytes and is required for early oocyte differentiation.It was found that miRNAs regulate target gene expression involved in follicle development,and miRNA-342-3p is expressed differentially in the plasma of patients with POI.By influencing transcription,post-transcriptional levels participate in regulating gene expression and affecting follicle function.In this project,the relationship and role of Sohlh1 and miRNA-342-3p and perimenopausal women in the study were taken to detect the levels of sex hormones in perimenopausal women,the differential expression of sohlhl gene regulation of transcriptional RNA and protein in the ovaries,and the differential expression of ovarian miRNA-342-3p in perimenopause women,so as to clarify the influence of Sohlh1 and miRNA-342-3p on the function of residual follicles of the ovaries in perimenopausal women.Mean:1.Patients who were treated in Shandong Provincial Hospital Affiliated to Shandong First Medical University were selected and divided into two groups according to age:perimenopausal women aged 48-53 as the experimental group;the control group were women of childbearing age aged 26-31.Collect 4 mL of fasting venous blood on the 3th day of menstruation,and instruct the patient to rest quietly for 15-30 minutes before blood collection.Blood samples were centrifuged at room temperature,and serum was collected and stored in a-80℃ refrigerator.Serum sex hormone levels were detected by ELISA,including anti-Müllerian hormone(AMH),follicle-stimulating hormone(FSH),luteinizing hormone(LH),and estradiol(E2).SPSS software was used to analyze the data2.Ovarian specimens were taken from patients undergoing ovarian surgery and divided into two groups according to age.After material sampling,fixation,dehydration,transparency,paraffin penetration,embedding,sectioning,hematoxylin and eosin staining,and sealing,the two groups were calculated respectively.The number of follicles in the whole section.3.Total RNA was extracted by Trizol method as a template for transcriptional synthesis of cDNA,the gene sequence of interest was retrieved through the GenBank database,primers were designed,fluorescence quantitative PCR amplification was performed,and the relative quantitative results of the target gene of each sample were calculated according to the original detection results of Real Time PCR and the relative quantitative calculation formula of 2-ΔΔCT method.4.Western blot method and immunohistochemistry detect the expression of Sohlh1 protein in ovarian tissue,and the obtained results are analyzed by image J image analysis software for protein gray value,and the expression of each histone is counted separately.Result:1.The endocrine data showed that the level of sex hormones in women perimenopausal women fluctuated sharply,FSH and LH were significantly increased,E2 and AMH were significantly reduced,The difference was statistically significant(P<0.05).menstrual patterns were irregular,and ovarian function decreased.HE staining of ovarian tissue slices shows a decrease in residual follicles,a decrease in oocytes,and an increase in ovarian fibrous tissue in perimenopausal women,The difference was statistically significant(P<0.05).2.The qPCR results showed that Sohlh1 was significantly reduced in mRNA expression in the ovaries of perimenopausal women,and miRNA-342-3p was significantly upregulated in the ovaries of perimenopausal women,The difference was statistically significant(P<0.05).3.In the Western-blot experiment,the Sohlh1 protein band was significantly downregulated compared to the control group,and protein grayscale analysis showed that The expression of Sohlh1 protein in the ovaries of perimenopausal women was reduced,The difference was statistically significant(P<0.05).4.Immunohistochemical experiments showed that the Sohlh1 protein is mainly expressed in the oocyte nucleus.It is expressed in large quantities in the nucleus of follicles,higher in granular cell layers and follicular membrane cells,and in small amounts in some interstitial cells.Compared with the control group,the expression of Sohlh1 protein in the follicles of perimenopausal women was significantly reduced,The difference was statistically significant(P<0.05).Conclusion:Serum sex hormone levels in perimenopausal women fluctuate violently,FSH and LH are generally elevated,E2 and AMH are reduced,and ovarian function is reduced,resulting in decreased reproductive function.In perimenopausal women,the residual follicles decreased,the ovarian reserve function decreased,and the Sohlh1 protein was expressed in the residual follicles of the ovaries of perimenopausal women,and the expression was reduced.The expression of miRNA-342-3p in the ovaries showed an upward trend,and according to the database,the target gene that miRNA-342-3p may be regulated,and it is speculated that Sohlh1 may be the target gene of miRNA-342-3p,which leads to follicle development,maturation disorders,and ultimately leads to decreased ovarian function.