Abstract
At low concentrations, cadmium (Cd) stimulates expression of cytoprotective genes. Cadmium induces the mRNA of NAD(P)H: quinone oxidoreductase 1 (NQO1) and heme oxygenase-1 (HO-1), which are nuclear factor (erythroid-derived 2)-like (Nrf2) transcription enzymes. Hence, the current study was designed to evaluate the probable protective role of iron (Fe) on Cd toxicity on the gonads of male rats. Twenty adult male albino rats were divided into four groups. The groups were designated as group A-control (rats exposed to exogenous Cd- and Fe-free water), group B (rats exposed to Cd-tainted water), group C (rats exposed to Fe-tainted water) and group D (rats exposed to Cd and Fe-tainted water concomitantly). The rats were initially gavaged with 0.229 mg/L Cd and 1.900 mg/L Fe tainted water, and the remaining solution was provided as drinking for the 4-week duration. Biochemical assays, hormone profiling, gene expression analysis, and histopathological evaluations were conducted on serum, plasma and testicular tissues. The study reveals that Cd induces oxidative stress, evidenced by decreased catalase (CAT) activities, whereas iron exposure showed antioxidant effects with alterations in malondialdehyde (MDA) levels. Concomitant exposure to Cd + Fe shows a reversal in the CAT activity in the serum of Cd-exposed rats. The levels of Luteinizing Hormone (LH) and Follicle-Stimulating Hormone (FSH) were significantly increased (p ≤ 0.05) in the plasma of rats exposed to Cd and Cd + Fe relative to the control. The testosterone and estradiol levels were significantly modulated in the plasma of rats exposed to Cd and Cd + Fe relative to the control. Exposure of rats to Cd only and Cd + Fe significantly downregulated HO-1 relative to the control. The mRNA level of NQO1 was significantly upregulated in the Cd only group relative to the control, but downregulated in the Cd + Fe group. The levels of Nrf2 and nuclear transcription factor kappa B (NF-κB) were upregulated in Cd only but downregulated in the Cd + Fe group relative to the control. Cadmium-induced alteration in mRNA levels in NQO1, Nrf2 and NF-κB was reversed in the presence of Fe. Histological changes in the testis of Cd-intoxicated rats were ameliorated in the presence of Fe. In conclusion, iron has the potential to offer protection against Cd-induced lesions in rat testis via a mechanism involving downregulation of NQO1/Nrf2 and NF-κB.
Data availability
The datasets generated and/or analysed during the current study are available on reasonable request from the corresponding author.
Abbreviations
- NQO1:
-
NAD(P)H: quinone oxidoreductase 1
- HO-1:
-
Heme oxygenase-1
- Nrf2:
-
Nuclear factor (erythroid-derived 2)-like factor-2
- NF-κB:
-
Nuclear transcription factor kappa B
- CAT:
-
Catalase
- MDA:
-
Malondialdehyde
- GSH:
-
Reduced glutathione
- SOD:
-
Superoxide dismutase
- LH:
-
Luteinizing Hormone
- FSH:
-
Follicle-Stimulating Hormone
- WT:
-
Wild type
- MEF:
-
Mouse Embryonic Fibroblasts
- ROS:
-
Reactive oxygen species
- DMT1:
-
Divalent metal transporter-1
- MTP1:
-
Metal transporter protein 1
- ELISA:
-
Enzyme-linked immunosorbent assay
- RNA:
-
Ribonucleic acid
- DNA:
-
Deoxyribonucleic acid
- cDNA:
-
Complementary Deoxyribonucleic acid
- PCR:
-
Polymerase chain reaction
- TRFC:
-
Transferrin receptor protein 1
- EPO:
-
Erythropoietin
- GAPDH:
-
Glycerol aldehyde phosphate dehydrogenase
References
Ogunbiyi, O. J., Iyare, H. E. & Apata, J. T. Toxic effects of cadmium and its association with iron on the liver and serum of wistar rats. Mintage J. Pharm. Med. Sci. 8 (3), 29–32 (2019).
Ogunbiyi, O. J. & Obi, F. O. Evaluation of cadmium toxicity and its association with iron on the gonads of female rats. Biokemistri 33 (3), 169–180 (2021).
Ogunbiyi, O. J. & Obi, F. O. Evaluation of gonadotoxic effects of cadmium and iron administered via tainted diet singly and combined in female rats. Toxicol. Risk Assess. 8 (1), 8pp. https://doi.org/10.23937/2572-4061.1510047 (2022).
Lawal, A. O. & Ellis, E. M. Nrf2-mediated adaptive response to cadmium-induced toxicity involves protein kinase C delta in human 1321N1 astrocytoma cells. Environ. Toxicol. Pharmacol. 32 (1), 54–62 (2011).
He, X., Chen, M. G. & Ma, Q. Activation of Nrf2 in defense against cadmium-induced oxidative stress. Chem. Res. Toxicol. 21 (7), 1375–1383 (2008).
Lawal, A. O. & Ellis, E. M. Differential sensitivity and responsiveness of three human cell lines HepG2, 1321N1, and HEK 293 to cadmium. J. Toxicol. Sci. 35 (4), 465–478 (2010).
Ryu, D. Y. et al. Dietary iron regulates intestinal cadmium absorption through iron transporters in rats. Toxicol. Letter. 152, 19–25 (2004).
Basha, D. C., Rani, M. U., Devi, C. B., Kumar, M. R. & Reddy, G. R. Perinatal lead exposure alters postnatal cholinergic and aminergic system in rat brain: Reversal effect of calcium co-administration. Int. J. Dev. Neurosci. 30, 343–350 (2012).
Djukić-Ćosić, D., Ninković, M., Maličević, Z., Matović, V. & Soldatović, D. Effect of magnesium pretreatment on reduced glutathione levels in tissues of mice exposed to acute and subacute cadmium intoxication: A time course study. Magnes. Res. 20, 177–186 (2007).
Egborge, A. B. M. Industrialization and heavy metal pollution in Warri River. 32nd Inaugural Lecture Series, University of Benin, Benin City, Nigeriap.31. (1991).
National Institute of Health (NIH). Guidelines for the Care and Use of Laboratory Animals no.85–93 (Revised NIH publication, 1985).
Allain, C. C., Poon, L., Chan, S. G., Richmond, W. & Fu, P. Enzymatic determination of total serum cholesterol. Clin. Chem. 20, 470 (1974).
Roeschlaw, P., Bernt, E. & Gruber, W. Enzymatic determination of total cholesterol in serum. J. Clin. Chem. Clin. Biochem. 12, 227 (1974).
Tietz, N. W. ED: Clinical Guide to Laboratory Tests 3rd edn (WA Saunders Co, 1995).
Buege, J. A. & Aust, S. D. Microsomal lipid peroxidation. Methods Enzymol. 52, 302 (1978). – 10.
Ellman, G. L. Tissue sulfhydryl groups. Arch. Biochem. Biophys. 82, 70–77 (1959).
Misra, H. P. & Fridovich, I. The role of superoxide anion in the autooxidation of epinephrine and a simple assay for superoxide dismutase. J. Biol. Chem. 247, 3170–3175 (1972).
Cohen, G., Dembiec, D. & Marcus, J. Measurement of catalase activity in tissue extracts. Anal. Biochem. 34, 30–38 (1970).
Elekofehinti, O. O., Onunkun, A. T. & Olaleye, M. T. Cymbopogon citratus (DC.) Stapf mitigates ER-stress induced by streptozotocin in rats via down-regulation of GRP78 and up-regulation of Nrf2 signaling. J. Ethnopharmacol. 262, 113–130 (2020).
Kiernan, J. A. Histological and histochemical methods. In: Theory and Practice. 4th ed. Bloxham, UK; pp. 51–55. (2008).
Maretta, M. & Marettova, E. Toxic Effects of Cadmium on the Female Reproductive Organs: A Review. Folia Vet. 66 (4), 56–66 (2022).
Jamakala, O. & Rani, A. U. Mitigating role of zinc and iron against cadmium induced toxicity in liver and kidney of male albino rat: a study with reference to metallothionine quantification. Int. J. Pharm. Pharm. Sci. 6 (9), 411–417 (2014).
Tribowo, J. A., Arizal, M. H., Nashrullah, M., Aditama, A. R. & Utama, D. G. Oxidative stress of cadmium-induced ovarian rat toxicity. Int. J. Chem. Eng. Appl. 5 (3), 254–258 (2014).
Filipic, M. Mechanisms of cadmium induced genomic instability. Mutat. Research/Fundamental Mol. Mechanism Mutagen. 733 (1–2), 69–77. https://doi.org/10.1016/j.mrfmmm.2011.09.002 (2012).
Sandalio, L. M., Dalurzo, H. C., Gomez, M., Romero, M. C. & Del-Rio, L. A. Cadmium-induced changes in the growth and oxidative metabolism of pea plants. J. Exp. Bot. 52 (364), 2115–2126 (2001).
Ogunbiyi, O. J. Evaluation of the effect of Benzene Contaminated Water on the Gravimetry, Serum Cholesterol and Triglyceride Concentrations of African Catfish (Clarias gariepinus). Mintage J. Pharm. Med. Sci. 6 (2), 12–14 (2017).
Nampoothiri, L. P., Agarwal, A. & Gupta, S. Effect of coexposure to lead and cadmium on antioxidant status in rat ovarian granulose cells. Archive Toxicol. 81, 145–150 (2007).
Priya, P. N., Pillai, A. & Gupta, S. Effect of simultaneous exposure to lead and cadmium on gonadotropin binding and steroidogenesis on granulosa cells: an in vitro study. Indian J. Exp. Biol. 42, 143–148 (2004).
Lafuente, A., Blanco, A., Ma´rquez, N., Alvarez-Demanuel, E. & Esquifino, A. I. Effects of acute and subchronic cadmium administration on pituitary hormone secretion in rat. Rev. Esp. Fisiologi. 53, 265–270 (1997).
Nawal, K. A., Ula, A. & Ban, T. S. Protective influence of zinc on reproductive parameters in male rat treated with cadmium. Am. J. Med. Med. Sci. 5 (2), 73–81 (2015).
De souza predes, F., Diamante, M. & Dolder, H. Testis response to low doses of cadmium in Wistar rats. Int. J. Exp. Pathol. 91, 125–131 (2010).
Li, H. & Wu, S. Current situation of research on toxicity of cadmium on the female gonad and reproduction. China Public. Health. 18, 379–381 (2002).
Ekhoye, E. I., Nwangwa, E. K. & Aloamaka, C. P. Changes in Some Testicular Biometric Parameters and Testicular Function in Cadmium Chloride Administered Wistar Rats. Br. J. Med. Med. Res. 3 (4), 2031–2041 (2013).
Koizumi, N., Murata, K., Hayashi, C., Nishio, H. & Goji, J. High cadmium accumulation among humans and primates: comparison across various mammalian species- a study from Japan. Biol. Trace Elem. Res. 121, 205–214 (2008).
Yang, P. M., Chen, H. C., Tsai, J. S. & Lin, L. Y. Cadmium induces Ca2+-dependent necrotic cell death through calpain-triggered mitochondrial depolarization and reactive oxygen species-mediated inhibition of nuclear factor-kappaB activity. Chem. Res. Toxicol. 20, 406–415 (2007).
van Raaij, S. E. G., Masereeuw, R., Swinkels, D. W. & van Swelm, R. P. M. Inhibition of Nrf2 alters cell stress induced by chronic iron exposure in human proximal tubular epithelial cells. Toxicol. Letter. 295, 179–186. https://doi.org/10.1016/j.toxlet.2018.06.1218 (2018).
Hassanein, E. H. M., Mohamed, W. R., Ahmed, O. S., Abdel-Daim, M. M. & Sayed, A. M. The role of inflammation in cadmium nephrotoxicity: NF-kb comes into view. Life Sci. 308, 120971. https://doi.org/10.1016/j.lfs.2022.120971 (2022).
Liu, T., Zhang, L., Joo, D. & Sun, S-C. NF-kb signalling in inflammation. Signal. Transduct. Target. Therapy. 2, e17023. https://doi.org/10.1038/sigtrans.2017.23 (2017).
Lu, Y. et al. Ketogenic diet attenuates oxidative stress and inflammation after spinal cord injury by activating Nrf2 and suppressing the NF-κB signaling pathways. Neurosci. Lett. 683, 13–18. https://doi.org/10.1016/j.neulet.2018.06.016 (2018).
Gammella, E., Buratti, P., Cairo, G. & Recalcati, S. The transferrin receptor: the cellular iron gate. Metallomics 9 (10), 1367–1375. https://doi.org/10.1039/c7mt00143f (2017).
Richard, C. & Verdier, F. Transferrin Receptors in Erythropoiesis. Int. J. Mol. Sci. 21 (24), 9713. https://doi.org/10.3390/ijms21249713 (2020).
Srai, S. K. et al. Erythropoietin regulates intestinal iron absorption in a rat model of chronic renal failure. Kidney Int. 78 (7), 660–667. https://doi.org/10.1038/ki.2010.217 (2010).
Ganz, T. Erythropoietin and iron- a conflicted alliance? Kidney Int. 78 (7), 660–667. https://doi.org/10.1016/j.kint.2018.07.027 (2018).
Kim, A. & Nemeth, E. New insights into iron regulation and erythropoiesis. Curr. Opin. Hematol. 22 (3), 199–205. https://doi.org/10.1097/MOH.0000000000000132 (2016).
Mouro, V. G. S. et al. Da Matta SLP. Subacute Testicular Toxicity to Cadmium Exposure Intraperitoneally and Orally. Oxidative Med. Cell. Longevity ;3429635. (2019).
Zirkin, B. R. & Papadopoulos, V. Leydig cells: formation, function, and regulation. Biol. Reprod. 99 (1), 101–111. https://doi.org/10.1093/biolre/ioy059 (2018).
Acknowledgements
The authors are grateful for the support received from the Petroleum Technology Development Fund (PTDF) with Ref No. PTDF/ED/ISS/PHD/OJO/1947/21.
We wish to acknowledge the support received from the management of Teady Bioscience Research Laboratory, Akure. Special thanks to Dr Akpeh Kingsley in the Biochemistry Department at the University of Benin for his assistance and support during this research.
Author information
Authors and Affiliations
Contributions
OOJ conceived the idea for this research and proposed the research design. OOJ and ONP supervised the research and were major contributors to writing the original manuscript. OOJ, EOO, and OFO conducted the experiment, analysed, and interpreted the research data. UVO performed the histological examination of the testes and interpreted the results. OOJ and ONP conducted a critical review of the revised manuscript and were major contributors to its editing. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethical approval
Ethical approval for the use of laboratory animals was obtained from the Faculty of Life Science Research Ethics Committee (FLSREC) of the University of Benin, with approval number FLSREC-2023-007.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Ogunbiyi, O.J., Okolie, N.P., Obi, F.O. et al. Protective effect of iron against cadmium-induced lesions in rat testis via downregulation of NQO1/Nrf2 and NF-κB. Sci Rep (2026). https://doi.org/10.1038/s41598-026-43876-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-43876-z