The Extremely-Low-Frequency Electromagnetic Field Affects Apoptosis and Oxidative-Stress-Related Genes and Proteins in the Porcine Endometrium-An In Vitro Study.

Nowadays, the extremely-low-frequency electromagnetic field (ELF-EMF) is recognized as environmental pollution. The data indicate that the ELF-EMF may affect factors related to epigenetic regulation and alter important biological processes in the uterus. The impact of the ELF-EMF on apoptosis and oxidative-stress-related genes has not been documented in porcine endometrium. This raises the question of whether the exposure to the ELF-EMF can induce apoptosis and/or oxidative stress in the endometrium of pigs during the peri-implantation period. Porcine endometrial slices (100 ± 5 mg) collected (n = 5) during the peri-implantation period were treated in vitro with ELF-EMF at a frequency of 50 Hz and flux density of 8 × 104 mG for 2 h. To determine the effect of ELF-EMF on apoptosis and oxidative stress in the endometrium, CASP3, CASP7, CIDEB, GADD45G, NOS1, NOS2, NOS3, and TP53I3 mRNA transcript were analyzed using real-time PCR, and protein abundance of CASP3, CASP7 using Western blot, and eNOS using ELISA were determined. Moreover, CASP3/7 and NOS activity was analyzed using flow cytometry and colorimetry, respectively. The decreased CASP7 and increased NOS3 mRNA transcript and protein abundance in ELF-EMF-treated endometrium were observed. Moreover, CIDEB, GADD45G, and TP53I3 mRNA transcript abundance was increased. Only p ≤ 0.05 was considered a statistically significant difference. The documented alterations indicate the potential of the ELF-EMF to affect apoptosis and generate oxidative stress in the endometrium. The insight into observed consequences documents for the first time the fact that the ELF-EMF may influence endometrial cell proliferation, angiogenesis, and/or tissue receptivity during peri-implantation.

Exposure to the extremely low-frequency electromagnetic field induces changes in the epigenetic regulation of gene expression in the endometrium.

Increasing technological development results in more sources of the extremely low-frequency electromagnetic field (ELF-EMF), which is recognized as an environmental risk factor. The results of the past study indicate that the ELF-EMF can affect the level of DNA methylation. The study aimed to determine whether the ELF-EMF induces changes in epigenetic regulation of gene expression in the endometrium of pigs during the peri-implantation period. Endometrial slices (100 ± 5 mg) collected on days 15-16 of pregnancy were exposed in vitro to the ELF-EMF at a frequency of 50 Hz for 2 h of treatment duration. To determine the impact of the ELF-EMF on elements of epigenetic regulations involved in DNA methylation, histone modification, and microRNA biogenesis in the endometrium, the DNMT1 and DNMT3a; EZH2, UHRF1, and MBD1; DICER1 and DGCR8 mRNA transcript and protein abundance were analyzed using Real-Time PCR and Western blot, respectively. Moreover, EED and SUZ12 mRNA transcript, global DNA methylation, and the activity of histone deacetylase (HDAC) were analyzed. The changes in the abundance of DNMT1 and DNMT3a, EZH2 mRNA transcript and protein, EED and SUZ12 mRNA transcript, global DNA methylation level, HDAC activity, and the abundance of proteins involved in microRNA biogenesis evoked by the ELF-EMF in the endometrium were observed. The ELF-EMF possesses the potential to alter epigenetic regulation of gene expression in the porcine endometrium. Observed alterations may be the reason for changes in the transcriptomic profile of the endometrium exposed to the ELF-EMF which in turn may disrupt biological processes in the uterus during peri-implantation.

Extremely low-frequency electromagnetic field exposure alters DNA methylation levels in the endometrium of pigs during the peri-implantation period.

CONTEXT: Extremely low-frequency electromagnetic field (ELF-EMF) emission is increasing due to substantial technological progress. The results of previous research provided evidence that ELF-EMF may exert changes in molecular mechanisms that control female reproduction. AIMS: We hypothesised that short-term ELF-EMF treatment alters the DNA methylation level of genes in the endometrium. Hence, the research aimed to determine the methylation level of selected genes whose expression was altered in response to ELF-EMF radiation in the endometrium of pigs during the peri-implantation period (days 15-16 of pregnancy). METHODS: Porcine endometrial slices (100±5mg) were collected during the peri-implantation period and exposed to ELF-EMF at a frequency of 50Hz for 2h in vitro . The control endometrium was not exposed to ELF-EMF. The level of DNA methylation in the promoter regions of EGR2 , HSD17B2 , ID2 , IL1RAP, MRAP2, NOS3, PTGER4, SERPINE1, VDR and ZFP57 was tested using qMS-PCR. KEY RESULTS: In the endometrium exposed to ELF-EMF, the level of methylation of HSD17B2 , MRAP2 , SERPINE1, VDR and ZFP57 was not altered; the level of methylation of EGR2 , ID2 and PTGER4 increased, and the level of methylation of IL1RAP and NOS3 decreased. CONCLUSIONS: ELF-EMF may alter the level of DNA methylation in the endometrium during the peri-implantation period. IMPLICATIONS: Changes in the DNA methylation induced by ELF-EMF may affect the transcriptomic profile of the endometrium and disturb physiological processes accompanying implantation and embryo development.

Extremely low-frequency electromagnetic field (ELF-EMF) induces alterations in epigenetic regulation in the myometrium - An in vitro study.

Previous research by the authors indicated that an extremely low-frequency electromagnetic field (ELF-EMF) evokes molecular alterations in the porcine myometrium. It was hypothesized that the ELF-EMF could induce alterations in the epigenetic regulation of gene expression in the myometrium. In the current study, slices of the porcine myometrium during the peri-implantation period (n = 4) were used for further in vitro exposition to ELF-EMF (50 Hz, 8 mT, 2 h treatment duration). The study tested whether the ELF-EMF may affect: 1/the expression of DNA (cytosine-5)-methyltransferase 1 (DNMT1) and DNA (cytosine-5)-methyltransferase 3a (DNMT3a), 2/the level of genomic DNA methylation, and 3/the level of amplification of methylated and unmethylated variants of promoter regions of selected genes with altered expression in response to ELF-EMF. It was found that ELF-EMF treatment increased DNMT1, decreased DNMT3a mRNA transcript and protein abundance, and increased the level of genomic DNA methylation. The direction of alterations in the level of amplification of methylated and unmethylated variants of the promoter region of selected genes with altered expression, i.e. prodynorphin (PDYN), interleukin 15 (IL15) signal transducer and activator of transcription 5A (STAT5A), tumor necrosis factor (TNF), and between down-regulated genes were early growth response 2 (EGR2), hyaluronan and proteoglycan link protein 1 (HAPLN1), and uteroferrin associated basic protein-2 (UABP2), mostly involving the direction of changes in their transcriptional activity, which was evaluated in a previous study by the authors. Thus, ELF-EMF radiation disturbs epigenetic mechanisms, which may underlay ELF-EMF-related transcriptomic alterations in the myometrium.

The effect of electromagnetic field (EMF) exposure on synthesis and release of steroid hormones by the porcine conceptuses during the peri-implantation period.

CONTEXT: Electrical devices and power systems are the sources of EM-waves which propagate everywhere in the environment. AIMS: The study aimed to determine whether EMF induced changes in the steroidogenesis of conceptuses and whether progesterone (P4 ) may be a possible protectant against the effects of EMF radiation. METHODS: The entire porcine conceptuses were collected during the peri-implantation period (days 15-16 of pregnancy), divided into fragments (100mg) and treated in vitro with EMF (50Hz or 120Hz, 2 or 4h exposure), and examined to determine of CYP17A1 , HSD3B1 , CYP19A3 , and HSD17B4 mRNA transcript and encoded protein abundance and the release of steroid hormones. Selected fragments of conceptuses were treated with P4 . KEY RESULTS: In conceptuses incubated without P4 , EMF at 120Hz decreased androstenedione (A4 ) and testosterone (T) release after 2h and increased oestrone (E1 ) release at 50Hz and 120Hz after 4h exposure. In P4 -treated conceptuses, EMF (50 and 120Hz, 4h exposure) decreased CYP19A3 mRNA transcript abundance, and increased (120Hz, 2h exposure) oestradiol-17ß (E2 ) release. CONCLUSIONS: The EMF radiation alters androgen and oestrogen synthesis and release from the conceptuses of pigs during the peri-implantation period. The P4 exerts protective effects on androgens and E1 release but it sensitises the conceptuses when comes to the mechanism of oestrogen synthesis and release during EMF radiation. IMPLICATIONS: The effect of EMF radiation on the steroidogenic pathway in conceptuses may induce disturbances in their proper development and implantation.

Effect of the Electromagnetic Field (EMF) Radiation on Transcriptomic Profile of Pig Myometrium during the Peri-Implantation Period-An In Vitro Study.

The electromagnetic field (EMF) affects the physiological processes in mammals, but the molecular background of the observed alterations remains not well established. In this study was tested the effect of short duration (2 h) of the EMF treatment (50 Hz, 8 mT) on global transcriptomic alterations in the myometrium of pigs during the peri-implantation period using next-generation sequencing. As a result, the EMF treatment affected the expression of 215 transcript active regions (TARs), and among them, the assigned gene protein-coding biotype possessed 90 ones (differentially expressed genes, DEGs), categorized mostly to gene ontology terms connected with defense and immune responses, and secretion and export. Evaluated DEGs enrich the KEGG TNF signaling pathway, and regulation of IFNA signaling and interferon-alpha/beta signaling REACTOME pathways. There were evaluated 12 differentially expressed long non-coding RNAs (DE-lnc-RNAs) and 182 predicted single nucleotide variants (SNVs) substitutions within RNA editing sites. In conclusion, the EMF treatment in the myometrium collected during the peri-implantation period affects the expression of genes involved in defense and immune responses. The study also gives new insight into the mechanisms of the EMF action in the regulation of the transcriptomic profile through lnc-RNAs and SNVs.

Electromagnetic Field (EMF) Radiation Alters Estrogen Release from the Pig Myometrium during the Peri-Implantation Period.

An electromagnetic field (EMF) may affect the functions of uterine tissues. This study hypothesized that EMF changes the estrogenic activity of pig myometrium during the peri-implantation period. Tissue was collected on days 15-16 of the gestation and incubated in the presence of EMF (50 and 120 Hz, 2 and 4 h). The cytochrome P450 aromatase type 3 (CYP19A3) and hydroxysteroid 17ß dehydrogenase type 4 (HSD17B4) mRNA transcript abundance, cytochrome P450arom (aromatase), and 17ß hydroxysteroid dehydrogenase 17ßHSD) protein abundance and estrone (E1) and estradiol-17ß (E2) release were examined using Real-Time PCR, Western blot and radioimmunoassay. Selected myometrial slices were treated with progesterone (P4) to determine whether it functions as a protector against EMF. CYP19A3 mRNA transcript abundance in slices treated with EMF was less at 50 Hz (2 h) and greater at 120 Hz (2 and 4 h). HSD17B4 mRNA transcript was greater in slices treated with EMF at 120 Hz (2 h). Progesterone diminished EMF-related effects on CYP19A3 and HSD17B4. When P4 was added, EMF had suppressive (50 and 120 Hz, 2 h) or enhancing (50 Hz, 4 h) effects on aromatase abundance. The E1 release was lower after 4 h of EMF treatment at 50 Hz and P4 did not protect myometrial E1 release. In conclusion, EMF alters the synthesis and release of E1 and did not affect E2 release in the myometrium during the peri-implantation period.