Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Activation of MIP-2 and MCP-5 Expression in Methylmercury-Exposed Mice and Their Suppression by N-Acetyl-L-Cysteine

  • 27 Accesses


Methylmercury (MeHg) is a well-known neurotoxin of the central nervous system (CNS). Neuroinflammation is one of the main pathways of MeHg-induced CNS impairment. This study aims to investigate the expressions of IL-6, MIP-2, and MCP-5, as biomarkers in relation with MeHg-induced CNS impairment and N-acetyl-L-cysteine (NAC) treatment in mice, as well as histopathological changes of brain tissue and clinical symptom such as ataxia. Twenty male Balb/c mice, aged 8–9 weeks, were divided into 4 groups and treated with saline (control), NAC [150 mg/kg body weight (BW) day], MeHg (4 mg Hg/kg BW), or a combination of MeHg and NAC for 17 days. MeHg induced the expression of IL-6, MIP-2, and MCP-5 in the serum, with median values (those in controls) of 55.06 (9.44), 15.94 (9.30), and 458.91 (239.91) mg/dl, respectively, and a statistical significance was observed only in IL-6 expression (p < 0.05). MIP-2 and MCP-5 expressions tended to increase in the cerebrum of MeHg-treated group compared with controls; however, the difference was not statistically significant. MeHg treatment also increased IL-6 expression in the cerebellum (7.73 and 4.81 mg/dl in MeHg-treated group and controls, respectively), with a marginal significance. NAC significantly suppressed MeHg-induced IL-6 and MIP-2 expressions in the serum (p < 0.05 for both), and slightly reduced MCP-5 expression in the cerebrum. Ataxia was observed in all MeHg-treated mice after 9-day exposure as well as the decrease of intact Purkinje cells in brain tissue (p < 0.05). These findings suggest that MeHg induced neurotoxicity by elevating the expression of IL-6, MIP-2, and MCP-5 and causing ataxia symptoms, and NAC reduced MeHg-mediated effects on the CNS.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3


  1. Aniszewska A, Chlodzinska N, Bartkowska K, Winnicka MM, Turlejski K, Djavadian RL (2015) The expression of interleukin-6 and its receptor in various brain regions and their roles in exploratory behavior and stress responses. J Neuroimmunol 284:1–9. https://doi.org/10.1016/j.jneuroim.2015.05.001

  2. Aremu DA, Madejczyk MS, Ballatori N (2008) N-acetylcysteine as a potential antidote and biomonitoring agent of methylmercury exposure. Environ Health Perspect 116:26–31. https://doi.org/10.1289/ehp.10383

  3. ATSDR (2017) ATSDR’s substance priority list. Agency for Toxic Substances and Disease Registry. https://www.atsdr.cdc.gov/spl/.

  4. Chang JY (2007) Methylmercury causes glial IL-6 release Neurosci Lett 416:217-220. https://doi.org/10.1016/j.neulet.2007.01.076

  5. Chang JY, Tsai PF (2009) IL-6 release from mouse glia caused by MeHg requires cytosolic phospholipase A2 activation. Neurosci Lett 461:85–89. https://doi.org/10.1016/j.neulet.2009.06.004

  6. Chen YW et al (2006) The role of phosphoinositide 3-kinase/Akt signaling in low-dose mercury-induced mouse pancreatic beta-cell dysfunction in vitro and in vivo. Diabetes 55:1614–1624. https://doi.org/10.2337/db06-0029

  7. David J, Nandakumar A, Muniroh M, Akiba S, Yamamoto M, Koriyama C (2017) Suppression of methylmercury-induced MIP-2 expression by N-acetyl-L-cysteine in murine RAW264.7 macrophage cell line. Eur J Med Res 22:45. https://doi.org/10.1186/s40001-017-0287-4

  8. Diab A et al (1999) Neutralization of macrophage inflammatory protein 2 (MIP-2) and MIP-1alpha attenuates neutrophil recruitment in the central nervous system during experimental bacterial meningitis. Infect Immun 67:2590–2601

  9. Eskes C, Honegger P, Juillerat-Jeanneret L, Monnet-Tschudi F (2002) Microglial reaction induced by noncytotoxic methylmercury treatment leads to neuroprotection via interactions with astrocytes and IL-6 release. Glia 37:43–52. https://doi.org/10.1002/glia.10019

  10. Farina M, Aschner M, Rocha JB (2011) Oxidative stress in MeHg-induced neurotoxicity Toxicol Appl Pharmacol 256:405–417. https://doi.org/10.1016/j.taap.2011.05.001

  11. Fowler MI, Weller RO, Heckels JE, Christodoulides M (2004) Different meningitis-causing bacteria induce distinct inflammatory responses on interaction with cells of the human meninges. Cell Microbiol 6:555–567. https://doi.org/10.1111/j.1462-5822.2004.00382.x

  12. Gadient RA, Otten U (1994) Expression of interleukin-6 (IL-6) and interleukin-6 receptor (IL-6R) mRNAs in rat brain during postnatal development. Brain Res 637:10–14. https://doi.org/10.1016/0006-8993(94)91211-4

  13. Godefroy D et al (2012) The chemokine CCL2 protects against methylmercury neurotoxicity. Toxicol Sci 125:209–218. https://doi.org/10.1093/toxsci/kfr252

  14. Gonzalez-Hernandez T et al (2006) Interleukin-6 and nitric oxide synthase expression in the vasopressin and corticotrophin-releasing factor systems of the rat hypothalamus. J Histochem Cytochem 54:427–441. https://doi.org/10.1369/jhc.5A6845.2005

  15. Guyenet SJ, Furrer SA, Damian VM, Baughan TD, La Spada AR, Garden GA (2010) A simple composite phenotype scoring system for evaluating mouse models of cerebellar ataxia. J Vis Exp. https://doi.org/10.3791/1787

  16. Hol J, Wilhelmsen L, Haraldsen G (2010) The murine IL-8 homologues KC, MIP-2, and LIX are found in endothelial cytoplasmic granules but not in Weibel-Palade bodies. J Leukoc Biol 87:501–508. https://doi.org/10.1189/jlb.0809532

  17. Hwang GW, Lee JY, Ryoke K, Matsuyama F, Kim JM, Takahashi T, Naganuma A (2011) Gene expression profiling using DNA microarray analysis of the cerebellum of mice treated with methylmercury. J Toxicol Sci 36:389–391. https://doi.org/10.2131/jts.36.389

  18. Kastenbauer S, Angele B, Sporer B, Pfister HW, Koedel U (2005) Patterns of protein expression in infectious meningitis: a cerebrospinal fluid protein array analysis J Neuroimmunol 164:134–139. https://doi.org/10.1016/j.jneuroim.2005.03.009

  19. Kim MS, Takahashi T, Lee JY, Hwang GW, Naganuma A (2012) Methylmercury induces CCL2 expression through activation of NF-kappaB in human 1321N1 astrocytes. J Toxicol Sci 37:1275–1278. https://doi.org/10.2131/jts.37.1275

  20. Klein M, Paul R, Angele B, Popp B, Pfister HW (2006) Koedel U, Protein expression pattern in experimental pneumococcal meningitis. Microbes Infect 8:974–983. https://doi.org/10.1016/j.micinf.2005.10.013

  21. Koedel U, Rupprecht T, Angele B, Heesemann J, Wagner H, Pfister HW, Kirschning CJ (2004) MyD88 is required for mounting a robust host immune response to Streptococcus pneumoniae in the CNS. Brain 127:1437–1445. https://doi.org/10.1093/brain/awh171

  22. Lee HY, Lee JS, Kim HG, Kim WY, Lee SB, Choi YH, Son CG (2017) The ethanol extract of Aquilariae Lignum ameliorates hippocampal oxidative stress in a repeated restraint stress mouse model. 17:397–BMC Complement Altern Med. https://doi.org/10.1186/s12906-017-1902-1

  23. Lee JY, Hwang GW, Kim MS, Takahashi T, Naganuma A (2012) Methylmercury induces a brain-specific increase in chemokine CCL4 expression in mice. J Toxicol Sci 37:1279–1282. https://doi.org/10.2131/jts.37.1279

  24. Luo Y, Fischer FR, Hancock WW, Dorf ME (2000) Macrophage inflammatory protein-2 and KC induce chemokine production by mouse astrocytes. J Immunol 165:4015–4023. https://doi.org/10.4049/jimmunol.165.7.4015

  25. Macedo-Junior SJ, Luiz-Cerutti M, Nascimento DB, Farina M, Soares Santos AR, de Azevedo Maia AH (2017) Methylmercury exposure for 14 days (short-term) produces behavioral and biochemical changes in mouse cerebellum, liver, and serum. J Toxicol Environ Health A 80:1145–1155. https://doi.org/10.1080/15287394.2017.1357324

  26. Mojsilovic-Petrovic J, Callaghan D, Cui H, Dean C, Stanimirovic DB, Zhang W (2007) Hypoxia-inducible factor-1 (HIF-1) is involved in the regulation of hypoxia-stimulated expression of monocyte chemoattractant protein-1 (MCP-1/CCL2) and MCP-5 (Ccl12) in astrocytes. J Neuroinflammation 4:12. https://doi.org/10.1186/1742-2094-4-12

  27. Moller AS, Bjerre A, Brusletto B, Joo GB, Brandtzaeg P, Kierulf P (2005) Chemokine patterns in meningococcal disease. J Infect Dis 191:768–775. https://doi.org/10.1086/427514

  28. Muniroh M, Khan N, Koriyama C, Akiba S, Vogel CF, Yamamoto M (2015) Suppression of methylmercury-induced IL-6 and MCP-1 expressions by N-acetylcysteine in U-87MG human astrocytoma cells. Life Sci 134:16–21. https://doi.org/10.1016/j.lfs.2015.04.024

  29. Peng S, Hajela RK, Atchison WD (2002) Characteristics of block by Pb2+ of function of human neuronal L-, N-, and R-type Ca2+ channels transiently expressed in human embryonic kidney 293 cells. Mol Pharmacol 62:1418–1430. https://doi.org/10.1124/mol.62.6.1418

  30. Roseguini BT, Silva LM, Polotow TG, Barros MP, Souccar C, Han SW (2015) Effects of N-acetylcysteine on skeletal muscle structure and function in a mouse model of peripheral arterial insufficiency. J Vasc Surg 61:777–786. https://doi.org/10.1016/j.jvs.2013.10.098

  31. Samuni Y, Goldstein S, Dean OM, Berk M (2013) The chemistry and biological activities of N-acetylcysteine. Biochim Biophys Acta 1830:4117–4129. https://doi.org/10.1016/j.bbagen.2013.04.016

  32. Sarafi MN, Garcia-Zepeda EA, MacLean JA, Charo IF, Luster AD (1997) Murine monocyte chemoattractant protein (MCP)-5: a novel CC chemokine that is a structural and functional homologue of human MCP-1. J Exp Med 185:99–109. https://doi.org/10.1084/jem.185.1.99

  33. Semple BD, Kossmann T, Morganti-Kossmann MC (2010) Role of chemokines in CNS health and pathology: a focus on the CCL2/CCR2 and CXCL8/CXCR2 networks. J Cereb Blood Flow Metab 30:459–473. https://doi.org/10.1038/jcbfm.2009.240

  34. Sirois JE, Atchison WD (2000) Methylmercury affects multiple subtypes of calcium channels in rat cerebellar granule cells Toxicol Appl Pharmacol 167:1–11. https://doi.org/10.1006/taap.2000.8967

  35. Spijker S (2011) Dissection of rodent brain regions. In, vol 57. pp 13–26. https://doi.org/10.1007/978-1-61779-111-6_2

  36. Takeuchi T, Eto K (1999) The pathology of Minamata disease : a tragic story of water pollution, with editorial collaboration of H. Nakayama and A. Sumiyoshi. Kyushu University Press, Inc., Fukuoka

  37. Whitney NP, Eidem TM, Peng H, Huang Y, Zheng JC (2009) Inflammation mediates varying effects in neurogenesis: relevance to the pathogenesis of brain injury and neurodegenerative disorders J Neurochem 108:1343–1359. https://doi.org/10.1111/j.1471-4159.2009.05886.x

  38. WHO (2017) Mercury and health. World Health Organization. https://www.who.int/en/news-room/fact-sheets/detail/mercury-and-health. Accessed 23 January 2019

  39. Wu Q et al. (2016) The Tibetan medicine Zuotai differs from HgCl2 and MeHg in producing liver injury in mice Regul Toxicol Pharmacol 78:1-7. https://doi.org/10.1016/j.yrtph.2016.03.017

  40. Yamamoto M, Khan N, Muniroh M, Motomura E, Yanagisawa R, Matsuyama T, Vogel CF (2017) Activation of interleukin-6 and -8 expressions by methylmercury in human U937 macrophages involves RelA and p50. J Appl Toxicol 37:611–620. https://doi.org/10.1002/jat.3411

  41. Yamamoto M, Motomura E, Yanagisawa R, Hoang VAT, Mogi M, Mori T, Nakamura M, Takeya M, Eto K (2019) Evaluation of neurobehavioral impairment in methylmercury-treated KK-Ay mice by dynamic weight-bearing test. J Appl Toxicol 39:221–230. https://doi.org/10.1002/jat.3710

  42. Yamamoto M, Yanagisawa R, Motomura E, Nakamura M, Sakamoto M, Takeya M, Eto K (2014) Increased methylmercury toxicity related to obesity in diabetic KK-Ay mice J Appl Toxicol 34:914–923. https://doi.org/10.1002/jat.2954

  43. Yasutake A, Hirayama K (1988) Sex and strain differences of susceptibility to methylmercury toxicity in mice. Toxicology 51:47–55. https://doi.org/10.1016/0300-483x(88)90079-0

Download references


This study was financially supported by Research Grant Number 38/UN.7.3.4/HK/2017 from Faculty of Medicine, Diponegoro University, Semarang, Central Java, Indonesia.

Author information

Correspondence to Muflihatul Muniroh.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Muniroh, M., Gumay, A.R., Indraswari, D.A. et al. Activation of MIP-2 and MCP-5 Expression in Methylmercury-Exposed Mice and Their Suppression by N-Acetyl-L-Cysteine. Neurotox Res (2020). https://doi.org/10.1007/s12640-020-00174-4

Download citation


  • Methylmercury
  • IL-6
  • MIP-2
  • MCP-5
  • N-acetyl-L-cysteine
  • Ataxia