Cell and Tissue Journal

Abstract Introduction: Numerous studies on microRNAs (miRNAs) have indicated that abnormal interactions between miRNAs and mRNAs contribute to the development and worsening of endometriosis. miR-223 influences genes associated with various biological functions such as signaling pathways, gene expression, cellular development, proliferation, angiogenesis, and programmed cell death. Alterations in genes have been demonstrated to be significant in the progression of endometriosis. The p53 gene, recognized as a tumor suppressor, plays a vital role in inhibiting cancer by regulating cell growth and division. Recent investigations suggest a potential connection between the p53 gene and endometriosis. In addition, Forkhead box protein O1 (FOXO1) acts as a cell-specific core transcription factor essential for effective endometrial remodeling throughout the menstrual cycle and may play an important role in the onset of endometriosis.
Aims: This study aimed to explore the expression levels of miR-223-3p and its target genes, TP53 and FOXO1, in cases of ovarian endometriosis through bioinformatic analysis and experimental verification.
Materials and methods: Gene expression data were obtained from the GSE105765 dataset in the Gene Expression Omnibus (GEO) database, which encompasses both normal (eutopic) and ectopic tissue samples from endometriosis patients. Differentially, expressed genes were pinpointed using the "limma" package in R software, with the results visualized as a heat map. Among the various expressed miRNAs, miR-223-3p was chosen for further investigation. Subsequently, the target genes of miR-223-3p were explored utilizing the miRDB database, leading to the identification of FOXO1 and TP53 as target genes. In the experimental component of this study, 40 ectopic tissue samples, 40 eutopic tissue samples from individuals with endometriosis, and 40 normal endometrial samples (control group) were analyzed. Following the extraction of total RNA and synthesis of cDNA, the expression levels of miR-223-3p, TP53, and FOXO1 were assessed using Real-Time PCR.
Results: The average relative expression of miR-223-3p in the ectopic group was measured at 4.13±0.72, while in the eutopic group it was 1.10±0.10, and in the control group it was 1.00±0.09. The relative expression of miR-223-3p was significantly higher in the ectopic group compared to both the eutopic and control groups (p<0.0001), whereas no significant differences were found between the control and eutopic groups (p=0.54). The average expression of FOXO1 in the ectopic group (0.47±0.12) was significantly lower than that in the eutopic group (1.02±0.06) and the control group (1.001±0.11). One-way ANOVA revealed a significant reduction in FOXO1 expression in the ectopic group when compared to the eutopic group and the control group (p<0.0001). However, there was no significant difference in FOXO1 expression between the eutopic and control groups (p=0.53). The relative expression level of the TP53 gene was analyzed in the ectopic group (0.335±0.14), the eutopic group (1.05±0.09), and the control group (1.01±0.12). Statistical evaluations showed that TP53 gene expression was significantly lower in the ectopic group compared to both the eutopic and control groups (p<0.0001). Conversely, no significant difference was observed in TP53 expression between the eutopic and control groups (p=0.28).
Conclusion: The findings indicate that miR-223-3p expression in ectopic samples was significantly elevated compared to both the eutopic and the control samples, while the expression of the FOXO1 and TP53 genes in the ectopic group was notably reduced relative to the eutopic and control groups. These results suggest that the expressions of miR-223-3p, TP53, and FOXO1 genes are changed in the endometrium of individuals with endometriosis, highlighting their potential roles in the disease's pathogenesis and their therapeutic implications.

Abstract Aims: In this study, the effects of Vitamin D3 on total protein concentration (TPC) and MBP expression in the corpus callosum extracts of Cuprizone induced demyelinated mouse has been investigated.
 Material and Methods: The mice were treated by Cuprizone for five weeks in order to induce demyelination. Then, the mice were divided into 3 groups. The first group was injected intraperitoneally (IP) by vitamin D3 in the amount of 5 µg/kg/daily body weight. The second group (SHAM) was injected IP by phosphate buffered saline (PBS) and the third group was left without injection as controls. After five weeks, the mice were killed and the corpus callosum was collected and the total protein concentration (TPC) and expression of MBP were studied by Western blot technique.
Results: No significant variation in the cortical TPC was seen in vitamin D3 injected group as compared to either SHAM or controls. We have also shown that the expression of MBP in the corpus callosum extracts of demyelinated mouse was significantly increased in the vitamin D3 injected group as compared to the other groups.
Conclusions: It is concluded that vitamin D3 increases MBP expression in the corpus callosum of cuprizone-induced demyelination mice. It is also suggested that vitamin D3 may play a role in the process of remyelination by increasing MBP expression in the cortex.
 
 

Abstract Aim: Infertility is the failure of a couple to engender after endeavoring at least one year of unprotected intercourse. male factor infertility accounts for approximately 50 percent of causes. Tumor necrosis factor-α (TNFα) is a multifunctional cytokine. TNF-α plays important role in the regulation of cellular processes related to spermatogenesis. There are two variants of the cell receptors that interacts with TNF-α. In the present study, the association of TNFR1 36A/G polymorphisms with idiopathic male infertility in the Guilan population was studied. Materials and Methods: This study consists of 106 infertile men and 114 fertile men as control group. Blood samples were taken and genomic DNA was extracted. Then genotypes and allele frequencies were assessed by PCR-RFLP method and the statistical analysis was performed by MedCalc software. Results: The frequencies of AA, AG, GG genotypes in patients were 41.5%, 7.5% and 50.9%, respectively and in controls were 45.6%, 33.3%, and 21.1%, respectively. The frequencies of A and G in patients were 0.45 and 0.55, respectively and in controls were 0.62 and 0.38, respectively. The results showed that there is a significant association between genotype frequency (p= 0.002) and allele frequency (p= 0.0004) in infertile and control groups. Conclusion: In conclusion, the subjects with G allele and GG genotype appears to be at greater risk of developing idiopathic infertility in Guilan province. Further studies with larger numbers of patients are required to elucidate the potential role of TNFR1 polymorphism in male infertility.