Abstract
Tungsten is widely used in medical, industrial, and military applications. The environmental exposure to tungsten has increased over the past several years, and few studies have addressed its potential toxicity. In this study, we evaluated the effects of chronic oral tungsten exposure (100 ppm) on renal inflammation in male mice. We found that 30- or 90-day tungsten exposure led to the accumulation of LAMP1-positive lysosomes in renal tubular epithelial cells. In addition, the kidneys of mice exposed to tungsten showed interstitial infiltration of leukocytes, myeloid cells, and macrophages together with increased levels of proinflammatory cytokines and p50/p65-NFkB subunits. In proximal tubule epithelial cells (HK-2) in vitro, tungsten induced a similar inflammatory status characterized by increased mRNA levels of CSF1, IL34, CXCL2, and CXCL10 and NFkB activation. Moreover, tungsten exposure reduced HK-2 cell viability and enhanced reactive oxygen species generation. Conditioned media from HK-2 cells treated with tungsten induced an M1-proinflammatory polarization of RAW macrophages as evidenced by increased levels of iNOS and interleukin-6 and decreased levels of the M2-antiinflammatory marker CD206. These effects were not observed when RAW cells were exposed to conditioned media from HK-2 cells treated with tungsten and supplemented with the antioxidant N-acetylcysteine (NAC). Similarly, direct tungsten exposure induced M1-proinflammatory polarization of RAW cells that was prevented by NAC co-treatment. Altogether, our data suggest that prolonged tungsten exposure leads to oxidative injury in the kidney ultimately leading to chronic renal inflammation characterized by a proinflammatory status in kidney tubular epithelial cells and immune cell infiltration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data supporting the findings of this study are available within the paper and its Supplementary Information.
References
Armstead AL, Li B. In vitro inflammatory effects of hard metal (WC-Co) nanoparticle exposure. Int J Nanomedicine. 2016;11:6195–206. https://doi.org/10.2147/IJN.S121141.
Bertinat R, Westermeier F, Silva P, Shi J, Nualart F, Li X, Yanez AJ. Anti-diabetic agent sodium tungstate induces the secretion of pro- and anti-inflammatory cytokines by human kidney cells. J Cell Physiol. 2017;232(2):355–62. https://doi.org/10.1002/jcp.25429.
Bolt AM, Mann KK. Tungsten: an emerging toxicant, alone or in combination. Curr Environ. Health Rep. 2016;3(4):405–15. https://doi.org/10.1007/s40572-016-0106-z.
Bolt AM, Sabourin V, Molina MF, Police AM, Negro Silva LF, Plourde D, Lemaire M, Ursini-Siegel J, Mann KK. Tungsten targets the tumor microenvironment to enhance breast cancer metastasis. Toxicol Sci. 2015;143(1):165–77. https://doi.org/10.1093/toxsci/kfu219.
Cheraghi G, Hajiabedi E, Niaghi B, Nazari F, Naserzadeh P, Hosseini MJ. High doses of sodium tungstate can promote mitochondrial dysfunction and oxidative stress in isolated mitochondria. J Biochem Mol Toxicol. 2019;33(4):e22266. https://doi.org/10.1002/jbt.22266.
Domingo JL. Vanadium and tungsten derivatives as antidiabetic agents: a review of their toxic effects. Biol Trace Elem Res. 2002, Aug;88(2):97–112. https://doi.org/10.1385/BTER:88:2:097.
Du H, Li Y, Wan D, Sun C, Sun J. Tungsten distribution and vertical migration in soils near a typical abandoned tungsten smelter. J Hazard Mater. 2022;429:128292. https://doi.org/10.1016/j.jhazmat.2022.128292.
Ezerina D, Takano Y, Hanaoka K, Urano Y, Dick TP. N-Acetyl cysteine functions as a fast-acting antioxidant by triggering intracellular H(2)S and sulfane sulfur production. Cell Chem Biol. 2018;25(4):447–459 e444. https://doi.org/10.1016/j.chembiol.2018.01.011.
Fox J, Macaluso F, Moore C, Mesenbring E, Johnson RJ, Hamman RF, James KA. Urine tungsten and chronic kidney disease in rural Colorado. Environ Res. 2021;195:110710. https://doi.org/10.1016/j.envres.2021.110710.
Grant MP, Henley N, Dubuissez M, Chen N, Hartmann U, Royal V, Barbier O, Pichette V, Gerarduzzi C. Subchronic oral exposure of tungsten induces myofibroblast transformation and various markers of kidney fibrosis. Am J Physiol Cell Physiol. 2022;322(2):C205–17. https://doi.org/10.1152/ajpcell.00277.2021.
Grant MP, VanderSchee CR, Chou H, Bolt A, Epure LM, Kuter D, Antoniou J, Bohle S, Mann KK, Mwale F. Tungsten accumulates in the intervertebral disc and vertebrae stimulating disc degeneration and upregulating markers of inflammation and pain. Eur Cell Mater. 2021;41:517–30. https://doi.org/10.22203/eCM.v041a33.
Guandalini GS, Zhang L, Fornero E, Centeno JA, Mokashi VP, Ortiz PA, Stockelman MD, Osterburg AR, Chapman GG. Tissue distribution of tungsten in mice following oral exposure to sodium tungstate. Chem Res Toxicol. 2011;24(4):488–93. https://doi.org/10.1021/tx200011k.
Hoffman JF, Vergara VB, Fan AX, Kalinich JF. Effect of embedded metal fragments on urinary metal levels and kidney biomarkers in the Sprague-Dawley rat. Toxicol Rep. 2021;8:463–80. https://doi.org/10.1016/j.toxrep.2021.02.023.
Jager KJ, Kovesdy C, Langham R, Rosenberg M, Jha V, Zoccali C. A single number for advocacy and communication-worldwide more than 850 million individuals have kidney diseases. Kidney Int. 2019;96(5):1048–50. https://doi.org/10.1016/j.kint.2019.07.012.
Jo SK, Sung SA, Cho WY, Go KJ, Kim HK. Macrophages contribute to the initiation of ischaemic acute renal failure in rats. Nephrol Dial Transplant. 2006;21(5):1231–9. https://doi.org/10.1093/ndt/gfk047.
Keith LS, Wohlers DW, Moffett DB, Rosemond ZA, Agency for Toxic, S., & Disease, R. ATSDR evaluation of potential for human exposure to tungsten. Toxicol Ind Health. 2007;23(5-6):309–45. https://doi.org/10.1177/0748233707081906.
Kelly AD, Lemaire M, Young YK, Eustache JH, Guilbert C, Molina MF, Mann KK. In vivo tungsten exposure alters B-cell development and increases DNA damage in murine bone marrow. Toxicol Sci. 2013;131(2):434–46. https://doi.org/10.1093/toxsci/kfs324.
Kirita Y, Wu H, Uchimura K, Wilson PC, Humphreys BD. Cell profiling of mouse acute kidney injury reveals conserved cellular responses to injury. Proc Natl Acad Sci U S A. 2020;117(27):15874–83. https://doi.org/10.1073/pnas.2005477117.
Ko GJ, Boo CS, Jo SK, Cho WY, Kim HK. Macrophages contribute to the development of renal fibrosis following ischaemia/reperfusion-induced acute kidney injury. Nephrol Dial Transplant. 2008;23(3):842–52. https://doi.org/10.1093/ndt/gfm694.
Kovago C, Szekeres B, Szucs-Somlyo E, Majlinger K, Jerzsele A, Lehel J. Preliminary study to investigate the distribution and effects of certain metals after inhalation of welding fumes in mice. Environ Sci Pollut Res Int. 2022;29(32):49147–60. https://doi.org/10.1007/s11356-022-19234-7.
Lalak NJ, Moussa SA. 'Corrosion' of tungsten spirale coils in the genitourinary tract. Clin Radiol. 2002;57(4):305–8. https://doi.org/10.1053/crad.2001.0791.
Lee S, Huen S, Nishio H, Nishio S, Lee HK, Choi BS, Ruhrberg C, Cantley LG. Distinct macrophage phenotypes contribute to kidney injury and repair. J Am Soc Nephrol. 2011;22(2):317–26. https://doi.org/10.1681/ASN.2009060615.
Lee SA, Noel S, Sadasivam M, Hamad ARA, Rabb H. Role of immune cells in acute kidney injury and repair. Nephron. 2017;137(4):282–6. https://doi.org/10.1159/000477181.
Lemus R, Venezia CF. An update to the toxicological profile for water-soluble and sparingly soluble tungsten substances. Crit Rev Toxicol. 2015;45(5):388–411. https://doi.org/10.3109/10408444.2014.1003422.
Liu YC, Zou XB, Chai YF, Yao YM. Macrophage polarization in inflammatory diseases. Int J Biol Sci. 2014;10(5):520–9. https://doi.org/10.7150/ijbs.8879.
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(4):402–8. https://doi.org/10.1006/meth.2001.1262.
Maekawa H, Inoue T, Ouchi H, Jao TM, Inoue R, Nishi H, Fujii R, Ishidate F, Tanaka T, Tanaka Y, Hirokawa N, Nangaku M, Inagi R. Mitochondrial damage causes inflammation via cGAS-STING signaling in acute kidney injury. Cell Rep. 2019;29(5):1261–1273 e1266. https://doi.org/10.1016/j.celrep.2019.09.050.
Mao L, Zheng L, You H, Ullah MW, Cheng H, Guo Q, Zhu Z, Xi Z, Li R. A comparison of hepatotoxicity induced by different lengths of tungsten trioxide nanorods and the protective effects of melatonin in BALB/c mice. Environ Sci Pollut Res Int. 2021;28(30):40793–807. https://doi.org/10.1007/s11356-021-13558-6.
Marui T, Tomonaga T, Izumi H, Yoshiura Y, Nishida C, Higashi H, Wang KY, Shijo M, Kubo M, Shimada M, Morimoto Y. Pulmonary toxicity of tungsten trioxide nanoparticles in an inhalation study and an intratracheal instillation study. J Occup Health. 2022;64(1):e12367. https://doi.org/10.1002/1348-9585.12367.
McCain WC, Crouse LC, Bazar MA, Roszell LE, Leach GJ, Middleton JR, Reddy G. Subchronic oral toxicity of sodium tungstate in Sprague-Dawley rats. Int J Toxicol. 2015;34(4):336–45. https://doi.org/10.1177/1091581815585568.
McWilliam SJ, Wright RD, Welsh GI, Tuffin J, Budge KL, Swan L, Wilm T, Martinas IR, Littlewood J, Oni L. The complex interplay between kidney injury and inflammation. Clin Kidney J. 2021;14(3):780–8. https://doi.org/10.1093/ckj/sfaa164.
Meng XM, Tang PM, Li J, Lan HY. Macrophage phenotype in kidney injury and repair. Kidney Dis (Basel). 2015;1(2):138–46. https://doi.org/10.1159/000431214.
Mihai S, Codrici E, Popescu ID, Enciu AM, Albulescu L, Necula LG, Mambet C, Anton G, Tanase C. Inflammation-related mechanisms in chronic kidney disease prediction, progression, and outcome. J Immunol Res. 2018;2018:2180373. https://doi.org/10.1155/2018/2180373.
Miller K, McVeigh CM, Barr EB, Herbert GW, Jacquez Q, Hunter R, Medina S, Lucas SN, Ali AS, Campen MJ, Bolt AM. Inhalation of tungsten metal particulates alters the lung and bone microenvironments following acute exposure. Toxicol Sci. 2021;184(2):286–99. https://doi.org/10.1093/toxsci/kfab109.
National Toxicology P. Toxicology and carcinogenesis studies of sodium tungstate dihydrate in Sprague Dawley (Hsd:Sprague Dawley SD) rats and B6C3F1/N mice (drinking water studies). Natl Toxicol Program Tech Rep Ser. 2021;599 https://doi.org/10.22427/NTP-TR-599.
O'Brown ZK, Van Nostrand EL, Higgins JP, Kim SK. The inflammatory transcription factors NFkappaB, STAT1 and STAT3 drive age-associated transcriptional changes in the human kidney. PLoS Genet. 2015;11(12):e1005734. https://doi.org/10.1371/journal.pgen.1005734.
Orr SE, Bridges CC. Chronic kidney disease and exposure to nephrotoxic metals. Int J Mol Sci. 2017;18(5) https://doi.org/10.3390/ijms18051039.
Roedel EQ, Cafasso DE, Lee KW, Pierce LM. Pulmonary toxicity after exposure to military-relevant heavy metal tungsten alloy particles. Toxicol Appl Pharmacol. 2012;259(1):74–86. https://doi.org/10.1016/j.taap.2011.12.008.
Rubin CS, Holmes AK, Belson MG, Jones RL, Flanders WD, Kieszak SM, Osterloh J, Luber GE, Blount BC, Barr DB, Steinberg KK, Satten GA, McGeehin MA, Todd RL. Investigating childhood leukemia in Churchill County, Nevada. Environ Health Perspect. 2007;115(1):151–7. https://doi.org/10.1289/ehp.9022.
Sachdeva S, Flora SJS. Efficacy of some antioxidants supplementation in reducing oxidative stress post sodium tungstate exposure in male wistar rats. J Trace Elem Med Biol. 2014;28(2):233–9. https://doi.org/10.1016/j.jtemb.2014.01.004.
Sachdeva S, Maret W. Comparative outcomes of exposing human liver and kidney cell lines to tungstate and molybdate. Toxicol Mech Methods. 2021;31(9):690–8. https://doi.org/10.1080/15376516.2021.1956031.
Sachdeva S, Sharma A, Flora SJS. MiADMSA abrogates sodium tungstate-induced oxidative stress in rats. Drug Chem Toxicol. 2022;45(6):2448–53. https://doi.org/10.1080/01480545.2021.1957560.
Shi N, Bai T, Wang X, Tang Y, Wang C, Zhao L. Toxicological effects of WS(2) nanomaterials on rice plants and associated soil microbes. Sci Total Environ. 2022;832:154987. https://doi.org/10.1016/j.scitotenv.2022.154987.
Tajima Y. Lacunary-substituted undecatungstosilicates sensitize methicillin-resistant Staphylococcus aureus to beta-lactams. Biol Pharm Bull. 2001, Sep;24(9):1079–84. https://doi.org/10.1248/bpb.24.1079.
Tajima Y. Effects of tungstosilicate on strains of methicillin-resistant Staphylococcus aureus with unique resistant mechanisms. Microbiol Immunol. 2003;47(3):207–12. https://doi.org/10.1111/j.1348-0421.2003.tb03388.x.
Tsai HJ, Wu PY, Huang JC, Chen SC. Environmental pollution and chronic kidney disease. Int J Med Sci. 2021;18(5):1121–9. https://doi.org/10.7150/ijms.51594.
Vervaet BA, Nast CC, Jayasumana C, Schreurs G, Roels F, Herath C, Kojc N, Samaee V, Rodrigo S, Gowrishankar S, Mousson C, Dassanayake R, Orantes CM, Vuiblet V, Rigothier C, D'Haese PC, De Broe ME. Chronic interstitial nephritis in agricultural communities is a toxin-induced proximal tubular nephropathy. Kidney Int. 2020;97(2):350–69. https://doi.org/10.1016/j.kint.2019.11.009.
Yilmaz MI, Carrero JJ, Axelsson J, Lindholm B, Stenvinkel P. Low-grade inflammation in chronic kidney disease patients before the start of renal replacement therapy: sources and consequences. Clin Nephrol. 2007;68(1):1–9. https://doi.org/10.5414/cnp68001.
Yu HH, Chen YC, Su HP, Chen L, Chen HH, Lin KA, Lin CH. Comparative pulmonary toxicity assessment of tungsten trioxide and tungsten trioxide hydrate nanoparticles. Sci Total Environ. 2023;855:158885. https://doi.org/10.1016/j.scitotenv.2022.158885.
Zhou P, Pan Y, Yuan B, Zhou J, Jiang J. Organ distribution of Nano-WC particles after repeated intratracheal instillation into the lungs of SD rats and subsequent organ injury. Biochem Biophys Res Commun. 2023;653:38–46. https://doi.org/10.1016/j.bbrc.2023.02.059.
Acknowledgements
We thank Dr. Koren K. Mann for providing invaluable feedback and thoughtful discussions on the manuscript.
Funding
This work was supported by grants from the Canadian Institutes of Health Research (CIHR Project Grant #428250 to C. Gerarduzzi), the Natural Sciences and Engineering Research Council of Canada (NSERC Discovery Grant #DGECR-2019-00141 to C. Gerarduzzi), Kidney Foundation of Canada (Kidney Health Research Grant #674586 to C.Gerarduzzi) and the Hôpital Maisonneuve-Rosemont Foundation (to C. Gerarduzzi). C. Gerarduzzi is a recipient of the Fonds de Recherche – Santé (Bourses de chercheurs-boursiers Junior 1 #298956) and the Support pour jeunes chercheur·euse·s: Mentorat et projet intercentre du réseau de recherche en santé Cardiométabolique, Diabète et Obésité (CMDO).
Author information
Authors and Affiliations
Contributions
Study design and conduct: C.G. and J.B.C. Data collection: J.B.C., N.H., M.G., and S.C. Data analysis and interpretation: C.G., J.B.C., M.G., P.G., and V.P. Drafting manuscript: J.B.C. and M.G. Revising manuscript content and approving the final version of manuscript: all authors reviewed the manuscript.
Corresponding author
Ethics declarations
Ethical approval
In vivo experiments were carried out according to the Canadian Council on Animal Care guidelines for the use of laboratory animals, under the supervision and approval of our local animal care committee (Comité de protection des animaux du Centre Intégré Universitaire de Santé et de Services Sociaux (CIUSSS) de l’Est-de-l’île-de-Montréal) with the approved protocol number 2018-1261, in compliance with the Animal Research: Reporting of in vivo Experiments (ARRIVE) Guidelines.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Barrera-Chimal, J., Henley, N., Grant, M. et al. Tungsten toxicity on kidney tubular epithelial cells induces renal inflammation and M1-macrophage polarization. Cell Biol Toxicol 39, 3061–3075 (2023). https://doi.org/10.1007/s10565-023-09817-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10565-023-09817-6