Iodothyronine deiodinase 2 (DiO2) regulates trophoblast cell line cycle, invasion and apoptosis; and its downregulation is associated with early recurrent miscarriage.

INTRODUCTION: Trophoblast development is a crucial event in placentation and pregnancy complications but its underlying mechanisms remain unclear. Thus, we aimed at investigating the role of DiO2 in trophoblast cell line decisions and assessing its placental villous expression in early recurrent miscarriage (ERM) patients. METHODS: The placental villous expression of DiO2 was determined with immunofluorescence. Cell proliferation was measured with the CCK8 kit while cell-cycle and apoptosis were studied with flow-cytometry. Cell migration and invasion were measured with wound-healing and transwell assays, respectively. Gene expression was then assessed with RT-qPCR and western blotting. RESULTS: DiO2 is expressed in the CTB, PCT, DCT and STB of the placenta. Its overexpression arrested trophoblast cell line proliferation at the G1 phase of the cell-cycle by downregulating cyclin-D1 and PCNA, while promoting apoptosis via increased caspase-3 activity and inhibition of the AKT and ERK1/2 signaling pathways. Also, it augmented trophoblast cell line migration and invasion via the upregulation of N-cadherin, vimentin, fascin-1, twist-1 and other epithelial-mesenchymal transition genes. DiO2 knockdown elicited the opposite effects. Surprisingly, each of these effects of DiO2 manipulation was not mediated by thyroid hormone metabolism. Assessment of the ERM placental villi revealed a downregulation of DiO2, N-cadherin, vimentin, fascin-1 and twist-1. The expression of E-cadherin remained unchanged in these placentae. DISCUSSION: During placentation, DiO2 may inhibit trophoblast proliferation while facilitating their differentiation into an invasive phenotype; and that its downregulation may contribute to the shallow trophoblast invasion that precedes ERM. Hence, DiO2 is a potential therapeutic target against ERM.

SNP rs12794714 of CYP2R1 is associated with serum vitamin D levels and recurrent spontaneous abortion (RSA): a case-control study.

PURPOSE: Vitamin D (VD) deficiency seems to be associated with the risk of recurrent spontaneous abortion (RSA). Vitamin D receptor (VDR) and cytochrome P450 family 2 subfamily R member 1 (CYP2R1) are two genes which are vital for VD metabolism and actions. However, whether single-nucleotide polymorphisms (SNPs) in these genes are correlated with the risk of RSA are poorly understood. Therefore, we aimed to characterize the relationships among VDR SNPs, CYP2R1 SNPs and RSA. METHODS: This case-control study enrolled 75 RSA patients and 83 controls. Serum VD and some cytokines were detected with LC-MS/MS and flow cytometry, respectively. Genotyping for three SNPs of CYP2R1 (rs10741657, rs10766197 and rs12794714) and five SNPs of VDR (rs7975232, rs1544410, rs2189480, rs2228570 and rs2239179) was done with polymerase chain reaction (PCR) and high-throughput sequencing. All the data were analyzed with appropriate methods and in different models. RESULTS: The results revealed a significant correlation between the AG genotype of CYP2R1 rs12794714 and VD levels (OR 0.686; 95% CI 0.49-0.96; p = 0.028). Besides, the AG and GG genotypes of CYP2R1 rs12794714 were markedly related to the risk of RSA (OR 52.394, 59.497; 95% CI 2.683-1023.265, 3.110-1138.367; p = 0.009, 0.007, respectively). CONCLUSION: Our results indicate that CYP2R1 rs12794714 might be a risk factor for RSA. Hence, early screening of pregnant women for CYP2R1 rs12794714 is necessary to warrant proactive counseling and treatment against RSA.

[Impact of cigarette smoking on sperm quality and seminal plasma ROS in preconception males].

OBJECTIVE: To investigate the influence of cigarette smoking on sperm quality and ROS in the seminal plasma of preconception males in Chongqing so as to provide some guidance for preconception couples. METHODS: Totally, 368 preconception males were enrolled in this study, including 196 smokers and 172 non-smokers, and the former divided into mild smokers (n = 88, ≤9 cigarettes per day for ≤5 years) and moderate to heavy smokers (n = 108, ≥10 cigarettes per day for ≥5 years or ≤9 cigarettes per day for ≥10 years). All the subjects underwent physical examination, medical history interview, health questionnaire investigation, and determination of semen parameters, ROS in the seminal plasma and sperm DNA fragmentation index (DFI). RESULTS: Among the 368 preconception males, 53.26% had a history of cigarette smoking, of whom 29.35% were moderate to heavy smokers. Only 55.4% of the subjects were found with normal sperm morphology and 52.6% with normal sperm progressive motility. Compared with the non-smokers, the moderate to heavy smokers showed significantly decreased semen volume (ï¼»3.33 ± 1.20ï¼½ vs ï¼»2.78 ± 1.08ï¼½ ml, P < 0.05), sperm concentration (ï¼»88.19 ± 70.33ï¼½ vs ï¼»75.16 ± 60.13ï¼½ × 106/ml, P < 0.05), and percentages of progressively motile sperm (PMS, ï¼»36.58 ± 13.90ï¼½ % vs ï¼»32.18 ± 15.24ï¼½ %, P < 0.05) and morphologically normal sperm (MNS, ï¼»3.85 ± 1.93ï¼½ % vs ï¼»3.52 ± 1.58ï¼½ %, P < 0.05), but increased sperm DFI (ï¼»10.45 ± 8.53ï¼½ % vs ï¼»14.53 ± 12.85ï¼½ %, P < 0.05) and ROS in the seminal plasma (ï¼»12.20 ± 8.10ï¼½ vs ï¼»24.10 ± 18.50ï¼½ nmol/mg prot, P < 0.05). Cigarette smoking was correlated positively with the ROS level in the seminal plasma (r = 0.235, P < 0.05), while the ROS level of the smokers negatively with the total sperm count (r = －136, P < 0.05), PMS (r = －0.381, P < 0.01) and MNS (r = －0.218, P < 0.01), but positively with sperm DFI in the preconception males (r = 0.387, P < 0.01). CONCLUSIONS: Cigarette smoking can increase the ROS level in the seminal plasma, decrease the total sperm count and sperm progressive motility, and induce sperm malformation and sperm DNA fragmentation in preconception males.