Immuno-histochemical localization of endothelial nitric oxide synthase in testicular cells of men with non-obstructive azoospermia

Background: Non obstructive azoospermia (NOA) is one of the causes of male infertility in which spermatogenesis process is disturbed. Recent studies suggested the possible role of endothelial nitric oxide synthase (eNOS) in spermatogenesis process. Objective: The aim of the present study is to evaluate the expression of eNOS in human testicular tissue in men with NOA and men with normal spermatogenesis by using immunocytochemistry. Materials and Methods: In this case-control study, testicular biopsies were obtained from 10 men with NOA and 7 men with normospermia who were attended to infertility center for diagnosis or infertility treatment. Immunohistochemistry was used to localize the isoform of eNOS in these tissues and the intensity of staining was semi quantitively assessed. In addition, the histopathological evaluation was examined in both groups. Results: The isoform of eNOS enzyme activity was detected in the cytoplasm of sertoli and leydig cells in both groups. There was, however, a considerable variability in the intensity of staining between two groups. The expression of eNOS in Leydig cells in control group was significantly (p<0.05) higher than those in the NOA group. In contrast, expression of eNOS in Sertoli cells in NOA was more than those in the control group. eNO Simmune staining was absent in the normal germ cells but was intense in the abnormal germ cells with piknotic neucleous. The most histopathological finding were hypospermatogenesis (27.2), Sertoli cell only syndrome (18.1) and tubular fibrotic (13.6). Conclusion: These results suggested that increase level of eNOS may play an important role in the apoptosis process in the abnormal germ cells and disturbance of spermatogenesis process

Expression profile and clonality of T-Cell receptor beta variable genes in normal human endometrium

Problem: In spite of their key immunological role, αβ+ T cells residing in endometrium have not been extensively explored. We analyzed here expression profile of TCRBV genes in normal human endometrium compared with peripheral blood. Methods: Samples were taken from normal reproductive women. RT-PCR using BV-gene specific primers was performed on blood and endometrial samples. After blotting, hybridization with radiolabelled probe and autoradiography, relative expression of each TCRBV family was determined. Clonal expansions of the over-expressed genes were assessed by CDR3 length polymorphism. Results: Only one gene (TCRBV7) was expressed significantly and two other genes marginally more in the endometrium compared with blood. All three TCRBV genes examined showed a rather restricted pattern in the endometrium in contrast to polyclonal patterns in the blood. Conclusion: Our results stress the similarities between T cells residing in different mucosal tissues and provide a basis for future investigations about endometrial T cells and their antigen specificities in gynecological problems. © 2006 The Authors Journal compilation 2006 Blackwell Munksgaard

Evaluation of Expression and Phosphorylation of Progesterone Receptor in Endometrial Stromal Cells of Patients with Recurrent Implantation Failure Compared to Healthy Fertile Women

Recurrent implantation failure (RIF) is the repeated failure of good-quality embryos in implantation process following several assisted reproduction cycles. Disruption of the endometrial receptivity is one of the main causes of RIF. Progesterone plays a pivotal role in the endometrial receptivity through the regulation of gene expression pattern by binding to its receptors in the endometrial cells. The aim of this study was to evaluate the expression level of progesterone receptor (PR) and its phosphorylated form in the endometrial stromal cells (eSC) of RIF patients and compare it to the eSC of healthy fertile women as control group. After isolation of the eSC from biopsy samples of RIF patients and healthy fertile women and their characterization, expression levels of PR mRNA, PR protein, and phospho-Ser294 PR protein were evaluated by quantitative real-time PCR and immunofluorescence staining, respectively. The results demonstrated a significant reduction in PR mRNA expression (P < 0.01.) and phospho-Ser294 PR protein (P < 0.05) level in RIF patients compared to the control group. These data for the first time suggest that the expression of PR and its phosphorylated form are impaired in RIF patients. Therefore, designing therapeutic methods for improving PR expression status and its regulation in the endometrium of RIF patients may help in improving the final reproductive outcomes of these cases. © 2021, Society for Reproductive Investigation

Evaluation of Expression and Phosphorylation of Progesterone Receptor in Endometrial Stromal Cells of Patients with Recurrent Implantation Failure Compared to Healthy Fertile Women

Recurrent implantation failure (RIF) is the repeated failure of good-quality embryos in implantation process following several assisted reproduction cycles. Disruption of the endometrial receptivity is one of the main causes of RIF. Progesterone plays a pivotal role in the endometrial receptivity through the regulation of gene expression pattern by binding to its receptors in the endometrial cells. The aim of this study was to evaluate the expression level of progesterone receptor (PR) and its phosphorylated form in the endometrial stromal cells (eSC) of RIF patients and compare it to the eSC of healthy fertile women as control group. After isolation of the eSC from biopsy samples of RIF patients and healthy fertile women and their characterization, expression levels of PR mRNA, PR protein, and phospho-Ser294 PR protein were evaluated by quantitative real-time PCR and immunofluorescence staining, respectively. The results demonstrated a significant reduction in PR mRNA expression (P < 0.01.) and phospho-Ser294 PR protein (P < 0.05) level in RIF patients compared to the control group. These data for the first time suggest that the expression of PR and its phosphorylated form are impaired in RIF patients. Therefore, designing therapeutic methods for improving PR expression status and its regulation in the endometrium of RIF patients may help in improving the final reproductive outcomes of these cases. © 2021, Society for Reproductive Investigation

The effect of pentoxifylline on the growth of endometrial implants and leukocytes in rats

Background: Immune system disturbances have an important role in endometriosis which may lead to infertility. It seems that inflammatory cytokines specially tumor necrosis factor alpha (TNF-α) which were produced by activated macrophages have an important role in pathology of endometriosis. Based on this theory, anti TNF-α drugs like pentoxifylline (PX) are suggested as new drugs for Endometriosis. Objective: This experimental study has been done on female rats to determine the effect of PX on the endometrial implants and leukocytes in serum. Material and Methods: In proestrous phase, one horn of rat's bicorn uterus was removed surgically and the endometrium was implanted to different places as follows: subcutaneous, peritoneum and near ovaries. After two months observation, female rats divided into two groups randomly. In treated group (n=10) PX (5mg/kg twice a day) and in control group (n=10), normal saline (same dose) were injected subcutaneously. Then, via second laparotomy and in the same phase of the cycles, the size of implants and the amount of leukocytes in serum were measured. Results: In treated group compared with control, the size of implants was decreased significantly in right subcutaneous (8.05mm vs 13.50mm) p<0.01, left subcutaneous (7.64 mm vs 14mm) p<0.01, right ovary (6.64 mm vs 15.22mm) p<0.001 and left ovary (7.18 mm vs 14.56 mm) p<0.005. In treated group, the total leukocyte count (5259.54 ± 178.78 vs 15833.33 ± 259.27) p<0.02 was decreased. The number of esterous cycle was similar in both groups. Conclusion: PX can reduce the size of endometrial implants as well as leukocyte count

Repertoire and clonality of t-cell receptor beta variable genes expressed in endometrium and blood t cells of patients with recurrent spontaneous abortion

Problem: Recurrent spontaneous abortion (RSA) is a relatively common disorder, the underlying causes of which are thought to be immunological in most cases. Method of Study: Expression profile and clonality pattern of T-cell receptor beta variable (TCRBV) genes in endometrium and blood of patients with RSA were investigated by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) using BV gene-specific primers. Relative expression of each BV family was determined and clonal expansion of the over-expressed genes was assessed by analysis of CDR3 length polymorphism. Results: Compared to blood, relative expression of four TCRBV genes was significantly higher in the endometrium of RSA group. Over-expressed genes, except for TCRBV3, all had restricted and oligoclonal patterns of expression in the endometrium. Conclusion: Endometrial T cells have a skewed TCRBV repertoire with restricted transcript heterogeneity, which is shared by both groups and minor variations observed in this pattern in RSA patients may reflect more recent and/or repeated exposure to nominal antigens or superantigens. © 2008 The Authors Journal compilation © 2008 Blackwell Munksgaard