Evaluation of Th17 and Treg cytokines in patients with unexplained recurrent pregnancy loss.

Background and Aim: The Th17/Treg balance in peripheral blood and reproductive tissues may have a role in the etiology of unexplained recurrent pregnancy loss (URPL). In this study, we evaluated the major cytokine of Treg cells transforming growth factor-beta (TGF-ß), and their specific transcription factor Forkhead box P3 (FOXP3), as well as the most highlighted cytokine of Th17 cells (interleukin [IL]-17A) in both URPL patients and healthy women. Methods: Samples were extracted from the peripheral blood, endocervix, endometrium, and vagina of 14 patients with URPL and 12 normal non-pregnant women as a control (normal) group. Quantitative reverse transcription polymerase chain reaction was used to determine gene expression. Enzyme-linked immunosorbent assay was used to determine the levels of cytokines in the serum and cervicovaginal fluid. Results: We found that in the URPL group, FOXP3 gene expression was considerably higher in peripheral blood than in the normal group (P=0.043). TGF-ß levels in the cervicovaginal fluid were different in the URPL and normal groups (P=0.015). In comparison to the control group, women with URPL had significantly greater IL-17 gene expression in the peripheral blood (P=0.01). Conclusion: Lower TGF-ß levels in the cervicovaginal fluid of patients compared to controls may be related with increased IL-17 and FOXP3 mRNA levels in patients with URPL. Dysregulation of local immune responses in reproductive tissues may represent dysregulation of systemic regulatory immunological responses in the pathogenesis of URPL. Relevance for Patients: Dysregulation of immune responses may play a role in the pathogenesis of URPL at least in some patients with URPL. We conclude that the breakdown of tolerance in the local immune responses is more critical than the breakdown of tolerance in systemic tolerance in the pathogenesis of URPL. Therefore, modulating immune responses in the endometrium and decidua may be the focus of future therapeutic approaches in URPL. The impact of seminal plasma on the expansion of Tregs may provide a novel therapeutic intervention that has already been used in assisted reproductive technologies. Therefore, we suggest that transvaginal TGF-ß in women with URPL may induce maternal tolerance which leads to the successful pregnancy.

Effects of vitamin D on steroidogenesis, reactive oxygen species production, and enzymatic antioxidant defense in human granulosa cells of normal and polycystic ovaries.

Polycystic ovary syndrome (PCOS) is accompanied with many disturbances in hormone synthesis and antioxidant defense. Previous reports have indicated that Vitamin D (vit.D) affects gene expression and have roles in normal follicular development. Therefore, we investigated the effects of vit.D on steroidogenesis, apoptosis, reactive oxygen species (ROS) production, and antioxidant defenses of human normal granulosa cells (N-GCs) and granulosa cells from polycystic ovaries (PCO-GCs). Ovarian GCs were obtained during oocyte retrieval procedure from 120 women with PCOS and from 100 healthy women who referred to Shiraz Fertility Center. The isolated GCs were cultured in the presence or absence of vit.D (100 nM), for 48 h. Concentration of sex steroids was measured by ELISA. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) expression and activities were assessed by q-PCR and photometric methods, respectively. The amount of ROS production was estimated using chemiluminescence and fluorescence methods. Cell viability and apoptosis were detected by Annexin-V/propidium iodide detection kit. Basal estrone and progesterone secretion by N-GCs was significantly higher than that of PCO-GCs. Vit.D significantly increased aromatase and 3ß-hydroxysteroid dehydrogenase activity in N-GCs and PCO-GCs. Basal expression and activity of GPx, in PCO-GCs were significantly lower than those of N-GCs. Treatment with vit.D significantly increased genes expression and enzyme activities in both groups. Basal ROS in PCO-GCs was markedly greater than that of N-GCs, which was attenuated by vit.D treatment. Cell apoptosis was directly correlated with ROS levels. We conclude that vit.D improved N-GCs and PCO-GCs functions through affecting steroidogenesis and enzymatic antioxidant defense. Under vit.D treatment, PCO-GCs could act more similar to N-GCs.

Influence of chemerin rs17173608 polymorphism on polycystic ovary syndrome susceptibility.

OBJECTIVE: The purpose of this study was to assess the association between the rs17173608 chemerin polymorphism and polycystic ovary syndrome risk (PCOS). MATERIALS AND METHODS: This case-control study was performed on 150 patients with PCOS and 150 normal women as the control group. Tetra-amplification refractory mutation system-polymerase chain reaction was used to detect the polymorphism. RESULTS: Our finding showed a positive association between the chemerin gene rs17173608 polymorphism and risk of PCOS. In the dominant effect of the G allele (comparison between TG+GG and TT), TG+GG genotypes were associated with the risk of PCOS (odds ratio = 2; 95% confidence interval = 1.3-3.2, p = 0.003). The G allele is thus dominant and increases the risk of PCOS as compared to the T allele (odds ratio = 1.7, 95% confidence interval = 1.1-2.5, p = 0.009). Nonetheless, there was no significant association between chemerin rs17173608 gene polymorphism and PCOS after adjusting genotypes for body mass index and age. CONCLUSION: These results suggested that there was a significant association between chemerin rs17173608 polymorphism and the PCOS; but this relationship was affected by obesity status.

Association between vaspin rs2236242 gene polymorphism and polycystic ovary syndrome risk.

Vaspin, an adipocytokine that has been isolated from the visceral adipose tissue, is a member of the serine protease inhibitor family. In humans, serum vaspin levels are correlated with body mass index (BMI) and obese women with polycystic ovary syndrome (PCOS). The present study is the first investigation to examine the association between vaspin rs2236242 gene polymorphism and risk of PCOS in Iranian patients. This case-control study was performed on 150 patients with PCOS and 150 healthy women. The vaspin genotypes were determined using tetra-amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR). Our finding showed that there are significant differences in genotype frequencies between case and control group regarding vaspin rs2236242 polymorphism (OR=0.59, CI=0.37-0.95, p=0.03). The A allele decreased the risk of PCOS (OR=0.67, CI=0.46-0.96, p=0.03) as compared to the T allele. There was no significant association between vaspin rs2236242 gene polymorphism and PCOS after adjusting genotypes for BMI. In conclusion, our data suggest a significant association between vaspin rs2236242 polymorphism and the PCOS but this relationship is affected by obesity status.