Radiofrequency Electromagnetic Field Exposure Amplifies the Detrimental Effects of Fetal Hyperglycemia in Zebrafish Embryos
Year 2024,
Volume: 83 Issue: 1, 77 - 84, 30.05.2024
Derya Cansız
,
Merih Beler
,
Gizem Eğilmezer
,
Semanur Işıkoğlu
,
Zülal Mızrak
,
İsmail Ünal
,
Selçuk Paker
,
Ahmet Ata Alturfan
,
Ebru Emekli Alturfan
Abstract
Objective: Radiofrequency electromagnetic field (RF-EMF) exposure during the embryonic period can cause defects in the development of the fetus. The study’s aim is to evaluate the effects of RF-EMF on the lipid accumulation, oxidant-antioxidant system parameters, locomotor activities, and gene expressions of insulin and leptin as genes related to insulin resistance in fetal hyperglycemia-induced zebrafish embryos.
Materials and Methods: The study exposed zebrafish embryos to RF-EMF (60 min) and glucose (5%) every day until 96 hours post fertilization (hpf). The study measured lipid peroxidation (LPO), superoxide dismutase, nitric oxide (NO), glutathione S-transferase (GST), and glutathione (GSH) levels to observe the oxidative stress status. The study monitored the development of the zebrafish embryos under a microscope, performed a locomotor activity analysis, measured acetylcholinesterase activity, and conducted oil red O staining to determine lipid accumulation. The study used reverse transcription polymerase chain reactions (RT-PCRs) to determine the expressions of ins and lepa by using RT-PCR.
Results: Both the glucose and RF-EMF applications decreased locomotor activity and increased the LPO and NO levels as oxidative damage indicators. Applying RF-EMF alone increased GST and GSH levels, while applying RF-EMF and glucose showed a decrease in the antioxidant defense systems. ins expression increased in the glucose and RF-EMF groups, while lepa expression increased in the glucose group and decreased in the RF-EMF group.
Conclusion: The harmful effects of hyperglycemia and RF-EMF exposure during the fetal period on embryo development need to be supported by studies to confirm the changes the current study has identified at the gene and protein levels.
Supporting Institution
Istanbul Medipol University
Thanks
On the 100th anniversary of the Republic of Turkey, we express our respect and endless gratitude to the great leader Mustafa Kemal Atatürk and his comrades in arms.
References
- Kashani ZA, Pakzad R, Fakari FR, et al. Electromagnetic fields exposure on fetal and childhood abnormalities: Systematic re-view and meta-analysis. Open Med (Wars). 2023;18(1):20230697. doi:10.1515/med-2023-0697 google scholar
- Scientific Committee on Emerging Newly Identified Health Risks. Opinion on potential health effects of exposure to electromagnetic fields. Bioelectromagnetics. 2015;36(6):480-484. google scholar
- Leszczynski D, Joenvâârâ S, Reivinen J, Kuokka R. Non-thermal activation of the hsp27/p38MAPK stress pathway by mobile phone radiation in human endothelial cells: molecular mechanism for cancer- and blood-brain barrier-related effects. Differentiation. 2002;70(2-3):120-129. google scholar
- Larsen AI, Olsen J, Svane O. Gender-specific reproductive out-come and exposure to high-frequency electromagnetic radia-tion among physiotherapists. Scand J Work Environ Health. 1991;17(5):324-329. google scholar
- Ouellet-Hellstrom R, Stewart WF. Miscarriages among fe-male physical therapists who report using radio- and microwave-frequency electromagnetic radiation. Am J Epidemiol. 1993;138(10):775-786. google scholar
- Rosas DB, Lopez H, Fernandez N. Is magnetic resonance imag-ing teratogenic during pregnancy? Literature review. Urologıa Colombiana. 2017;26(3):219-228. google scholar
- Kaplan G, Beler M, Unal I, et al. Diethylhexyl phthalate expo-sure amplifies oxidant and inflammatory response in fetal hy-perglycemia model predisposing insulin resistance in zebrafish embryos. Toxicol Ind Health. 2024;40(5):232-243. google scholar
- Zang L, Maddison LA, Chen W. Zebrafish as a model for obesity and diabetes. Front Cell Dev Biol. 2018;20(6):91. doi:10.3389/fcell.2018.00091 google scholar
- 9. Rocha F, Dias J, Engrola S, et al. Glucose metabolism and gene expression in juvenile zebrafish (Danio rerio) challenged with a high carbohydrate diet: effects of an acute glucose stimulus during late embryonic life. Br JNutr. 2015;113:403-413. google scholar
- Gut P, Baeza-Raja B, Andersson O, et al. Whole-organism screening for gluconeogenesis identifies activators of fasting metabolism. Nat Chem Biol. 2013; 9:97-104. google scholar
- Hansen NS, Strasko KS, Hjort L, et al. Fetal hyper-glycemia changes human preadipocyte function in adult life. J Clin Endocrinol Metab. 2017;102(4):1141-1150. google scholar
- Westerfield M.The zebrafish Book. Guide for the laboratory use of zebrafish (Danio rerio). University of Oregon Press, Eugene. 1995. google scholar
- Cansız D, Unal I, Beler M, Alturfan AA, Emekli-Alturfan E. As-sessment of developmental neurotoxicity using semi-automatic behavior analysis system for zebrafish. Methods Mol Biol. 2024;2753:409-419. google scholar
- Lowry OH, Rosebrough NJ, Farr A., Randall RJ. Protein measure-ment with the Folin phenol reagent. J Biol Chem. 1951;193:265-275. google scholar
- Yagi K. Simple assay for the level of total lipid peroxides in serum or plasma. Methods Mol Biol 1998;108:101-106. google scholar
- Miranda KM, Espey MG, Wink DA. A rapid, simple spectropho-tometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide. 2001;5:62-71. google scholar
- Habig WH, Jakoby WB. Assays for differentiation of glutathione S-transferases. Methods Enzymol. 1981;77:398-405. google scholar
- Mylroie AA, Collins H, Umbles C, Kyle J. Erythrocyte superoxide dismutase activity and other parameters of copper status in rats ingesting lead acetate. Toxicol Appl Pharmacol. 1986;82:512-520. google scholar
- Ellman GL, Courtney KD, Andres Jr V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88-95. google scholar
- Beutler E, Duron O, Kelly BM. Improved method for the determi-nation of blood glutathione. J Lab Clin Med. 1963;61:882-888. google scholar
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method Methods. 2001;25:402-408. google scholar
- Kim SH, Wu SY, Baek JI, Choi SY, Su Y, Flynn CR, et al. A post-developmental genetic screen for zebrafish models of inherited liver disease. PLoS ONE. 2015;10(5): e0125980. doi:10.1371/journal.pone.0125980 google scholar
- Mallozzi M, Bordi G, Garo C, Caserta D. The effect of maternal exposure to endocrine disrupting chemicals on fetal and neonatal development: A review on the major concerns. Birth Defects Res C Embryo Today. 2016;108(3);224-242. google scholar
- Singh A, Castillo HA, Brown J, Kaslin J, Dwyer KM, Gibert Y. High glucose levels affect retinal patterning during zebrafish embryogenesis. Sci Rep. 2019;11(1):4121. doi:10.1038/s41598-019-41009-3 google scholar
- Li Y, Chen Q, Liu Y, et al. High glucose-induced ROS-accumulation in embryo-larval stages of zebrafish leads to mitochondria-mediated apoptosis. Apoptosis. 2022;27: 509-520. google scholar
- Guru A and Arockiaraj J. Exposure to environmental pollutant bisphenol A causes oxidative damage and lipid accumulation in Zebrafish larvae: Protective role of WL15 peptide derived from cysteine and glycine-rich protein 2. J Biochem Mol Toxicol. 2023;37(1):e23223. doi:10.1002/jbt.23223 google scholar
- Erion DM, Park HJ, Lee HY. The role of lipids in the pathogenesis and treatment of type 2 diabetes and associated co-morbidities. BMB Rep. 2016;49(3):139-148. google scholar
- Morris S, Cholan PM, Britton WJ et al. Glucose inhibits haemostasis and accelerates diet-induced hyperlipidaemia in ze-brafish larvae. Sci Rep. 2021;11(1):19049. doi:10.1038/s41598-021-98566-9 google scholar
- Agha-Hosseini F, Mirzaii-Dizgah I, Farmanbar N, Abdollahi M. Oxidative stress status and DNA damage in saliva of human sub-jects with oral lichen planus and oral squamous cell carcinoma. J Oral Pathol Med. 2012; 41:736-740. google scholar
- Torres-Ruiz M, Suarez Vi. Lopez OJ, et al. Effects of 700 and 3500 MHz 5G radiofrequency exposure on develop-ing zebrafish embryos. Sci Total Environ. 2024;915:169475. doi:10.1016/j.scitotenv.2023.169475 google scholar
- Briaud I, Rouault C, Bailbe D, Portha B, Reach G, Poitout V. Glucose-induced insulin mRNA accumulation is impaired in islets from neonatal streptozotocin-treated rats. Horm Metab Res. 2000;32(3):103-106. google scholar
- Meo SA and Rubeaan KA. Effects of exposure to electromagnetic field radiation (EMFR) generated by activated mobile phones on fasting blood glucose. UOMEH. 2013;26:235-241. google scholar
- Al-Hussaniy HA, Alburghaif AH, Naji MA. Leptin hormone and its effectiveness in reproduction, metabolism, immunity, diabetes, hopes and ambitions. JMed Life. 2021;14:600-605. google scholar
- Kelesidis T, Mantzoros CS. The emerging role of leptin in hu-mans. Pediatr Endocrinol Rev. 2006;3(3):239-248. google scholar
Year 2024,
Volume: 83 Issue: 1, 77 - 84, 30.05.2024
Derya Cansız
,
Merih Beler
,
Gizem Eğilmezer
,
Semanur Işıkoğlu
,
Zülal Mızrak
,
İsmail Ünal
,
Selçuk Paker
,
Ahmet Ata Alturfan
,
Ebru Emekli Alturfan
References
- Kashani ZA, Pakzad R, Fakari FR, et al. Electromagnetic fields exposure on fetal and childhood abnormalities: Systematic re-view and meta-analysis. Open Med (Wars). 2023;18(1):20230697. doi:10.1515/med-2023-0697 google scholar
- Scientific Committee on Emerging Newly Identified Health Risks. Opinion on potential health effects of exposure to electromagnetic fields. Bioelectromagnetics. 2015;36(6):480-484. google scholar
- Leszczynski D, Joenvâârâ S, Reivinen J, Kuokka R. Non-thermal activation of the hsp27/p38MAPK stress pathway by mobile phone radiation in human endothelial cells: molecular mechanism for cancer- and blood-brain barrier-related effects. Differentiation. 2002;70(2-3):120-129. google scholar
- Larsen AI, Olsen J, Svane O. Gender-specific reproductive out-come and exposure to high-frequency electromagnetic radia-tion among physiotherapists. Scand J Work Environ Health. 1991;17(5):324-329. google scholar
- Ouellet-Hellstrom R, Stewart WF. Miscarriages among fe-male physical therapists who report using radio- and microwave-frequency electromagnetic radiation. Am J Epidemiol. 1993;138(10):775-786. google scholar
- Rosas DB, Lopez H, Fernandez N. Is magnetic resonance imag-ing teratogenic during pregnancy? Literature review. Urologıa Colombiana. 2017;26(3):219-228. google scholar
- Kaplan G, Beler M, Unal I, et al. Diethylhexyl phthalate expo-sure amplifies oxidant and inflammatory response in fetal hy-perglycemia model predisposing insulin resistance in zebrafish embryos. Toxicol Ind Health. 2024;40(5):232-243. google scholar
- Zang L, Maddison LA, Chen W. Zebrafish as a model for obesity and diabetes. Front Cell Dev Biol. 2018;20(6):91. doi:10.3389/fcell.2018.00091 google scholar
- 9. Rocha F, Dias J, Engrola S, et al. Glucose metabolism and gene expression in juvenile zebrafish (Danio rerio) challenged with a high carbohydrate diet: effects of an acute glucose stimulus during late embryonic life. Br JNutr. 2015;113:403-413. google scholar
- Gut P, Baeza-Raja B, Andersson O, et al. Whole-organism screening for gluconeogenesis identifies activators of fasting metabolism. Nat Chem Biol. 2013; 9:97-104. google scholar
- Hansen NS, Strasko KS, Hjort L, et al. Fetal hyper-glycemia changes human preadipocyte function in adult life. J Clin Endocrinol Metab. 2017;102(4):1141-1150. google scholar
- Westerfield M.The zebrafish Book. Guide for the laboratory use of zebrafish (Danio rerio). University of Oregon Press, Eugene. 1995. google scholar
- Cansız D, Unal I, Beler M, Alturfan AA, Emekli-Alturfan E. As-sessment of developmental neurotoxicity using semi-automatic behavior analysis system for zebrafish. Methods Mol Biol. 2024;2753:409-419. google scholar
- Lowry OH, Rosebrough NJ, Farr A., Randall RJ. Protein measure-ment with the Folin phenol reagent. J Biol Chem. 1951;193:265-275. google scholar
- Yagi K. Simple assay for the level of total lipid peroxides in serum or plasma. Methods Mol Biol 1998;108:101-106. google scholar
- Miranda KM, Espey MG, Wink DA. A rapid, simple spectropho-tometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide. 2001;5:62-71. google scholar
- Habig WH, Jakoby WB. Assays for differentiation of glutathione S-transferases. Methods Enzymol. 1981;77:398-405. google scholar
- Mylroie AA, Collins H, Umbles C, Kyle J. Erythrocyte superoxide dismutase activity and other parameters of copper status in rats ingesting lead acetate. Toxicol Appl Pharmacol. 1986;82:512-520. google scholar
- Ellman GL, Courtney KD, Andres Jr V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88-95. google scholar
- Beutler E, Duron O, Kelly BM. Improved method for the determi-nation of blood glutathione. J Lab Clin Med. 1963;61:882-888. google scholar
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method Methods. 2001;25:402-408. google scholar
- Kim SH, Wu SY, Baek JI, Choi SY, Su Y, Flynn CR, et al. A post-developmental genetic screen for zebrafish models of inherited liver disease. PLoS ONE. 2015;10(5): e0125980. doi:10.1371/journal.pone.0125980 google scholar
- Mallozzi M, Bordi G, Garo C, Caserta D. The effect of maternal exposure to endocrine disrupting chemicals on fetal and neonatal development: A review on the major concerns. Birth Defects Res C Embryo Today. 2016;108(3);224-242. google scholar
- Singh A, Castillo HA, Brown J, Kaslin J, Dwyer KM, Gibert Y. High glucose levels affect retinal patterning during zebrafish embryogenesis. Sci Rep. 2019;11(1):4121. doi:10.1038/s41598-019-41009-3 google scholar
- Li Y, Chen Q, Liu Y, et al. High glucose-induced ROS-accumulation in embryo-larval stages of zebrafish leads to mitochondria-mediated apoptosis. Apoptosis. 2022;27: 509-520. google scholar
- Guru A and Arockiaraj J. Exposure to environmental pollutant bisphenol A causes oxidative damage and lipid accumulation in Zebrafish larvae: Protective role of WL15 peptide derived from cysteine and glycine-rich protein 2. J Biochem Mol Toxicol. 2023;37(1):e23223. doi:10.1002/jbt.23223 google scholar
- Erion DM, Park HJ, Lee HY. The role of lipids in the pathogenesis and treatment of type 2 diabetes and associated co-morbidities. BMB Rep. 2016;49(3):139-148. google scholar
- Morris S, Cholan PM, Britton WJ et al. Glucose inhibits haemostasis and accelerates diet-induced hyperlipidaemia in ze-brafish larvae. Sci Rep. 2021;11(1):19049. doi:10.1038/s41598-021-98566-9 google scholar
- Agha-Hosseini F, Mirzaii-Dizgah I, Farmanbar N, Abdollahi M. Oxidative stress status and DNA damage in saliva of human sub-jects with oral lichen planus and oral squamous cell carcinoma. J Oral Pathol Med. 2012; 41:736-740. google scholar
- Torres-Ruiz M, Suarez Vi. Lopez OJ, et al. Effects of 700 and 3500 MHz 5G radiofrequency exposure on develop-ing zebrafish embryos. Sci Total Environ. 2024;915:169475. doi:10.1016/j.scitotenv.2023.169475 google scholar
- Briaud I, Rouault C, Bailbe D, Portha B, Reach G, Poitout V. Glucose-induced insulin mRNA accumulation is impaired in islets from neonatal streptozotocin-treated rats. Horm Metab Res. 2000;32(3):103-106. google scholar
- Meo SA and Rubeaan KA. Effects of exposure to electromagnetic field radiation (EMFR) generated by activated mobile phones on fasting blood glucose. UOMEH. 2013;26:235-241. google scholar
- Al-Hussaniy HA, Alburghaif AH, Naji MA. Leptin hormone and its effectiveness in reproduction, metabolism, immunity, diabetes, hopes and ambitions. JMed Life. 2021;14:600-605. google scholar
- Kelesidis T, Mantzoros CS. The emerging role of leptin in hu-mans. Pediatr Endocrinol Rev. 2006;3(3):239-248. google scholar