Archives of Toxicology

, Volume 88, Issue 2, pp 323–336 | Cite as

ERK pathway is activated in bare-FeNPs-induced autophagy

Molecular Toxicology

Abstract

Iron oxide nanoparticles (FeNPs) are known to be one of the most biocompatible and safe nanoparticles. However, their long-term persistence remains a problem, and macrophages play as an important mediator in continuous stimulation of the immune system due to biopersistence of nanoparticles. In the present study, we identified the mechanisms underlying the uptake and toxicity of bare-FeNPs using RAW264.7 cells, a mouse peritoneal macrophage cell line. The bare-FeNPs penetrated the cell membrane through electrostatic interactions together with the general phagocytic pathway. At 24 h after exposure, they distributed freely in the cytosol or within autophagosome-like vacuoles. Bare-FeNPs induced decrease in the cell viability along with the cell cycle arrest in G1 phase. In addition, they increased the generation of ROS and the secretion of NO and TNF alpha as well as the expression of SOD-1 and SOD-2 proteins, which are an antioxidant. While the mitochondrial calcium level, the intensity of labeled mitochondria, and ATP production decreased, the levels of autophagy-related proteins such as p62, beclin 1, ATG5, and LC3B increased in a dose-dependent manner together with the levels of ATF 3, p-EGFR, and p-ERK proteins. However, the level of p-JNK protein clearly decreased. TEM images also showed that damaged organelle exist within autophagosome-like vacuoles with bare-FeNPs. On the basis of these results, we suggest that bare-FeNPs induce autophagy by initiating oxidative stress in RAW264.7 cells. Furthermore, ERK, but not JNK, pathway is activated in bare-FeNPs-induced autophagy.

Keywords

Iron oxide nanoparticles Macrophage Autophagy ERK Mitochondria 

Supplementary material

204_2013_1134_MOESM1_ESM.ppt (408 kb)
EDS analysis of cell exposed to bare-FeNPs. (a) bright field and (b) dark field, and (d) EDS spectra of star point in TEM image (PPT 408 kb)
204_2013_1134_MOESM2_ESM.docx (21 kb)
Supplementary material 2 (DOCX 21 kb)
204_2013_1134_MOESM3_ESM.ppt (264 kb)
Decrease in pH following bare-FeNPs exposure. The pH value in cell culture media was measured within maximum 5 min after incubation finishing. The experiment was performed three times, independently, and the value means AV ± SD (PPT 263 kb)

References

  1. Buyukhatipoglu K, Clyne AM (2011) Superparamagnetic iron oxide nanoparticles change endothelial cell morphology and mechanics of reactive oxygen species formation. J Biomed Master Res A 96:186–195CrossRefGoogle Scholar
  2. Castaneda RT, Khurana A, Khan R, Daldrup-Link HE (2011) Labeling stem cells with ferumoxytol, an FDA-approved iron oxide nanoparticle. J Vis Exp 57:e3482PubMedGoogle Scholar
  3. Chang L, Karin M (2001) Mammalian MAP kinase signaling cascades. Nature 410:37–40PubMedCrossRefGoogle Scholar
  4. Chen Y, Chen BA (2010) Application and advancement of magnetic iron-oxide nanoparticles in tumor-targeted therapy. Chin J Cancer 29:125–128PubMedGoogle Scholar
  5. Davis RJ (2000) Signal transduction by the JNK group of MAP kinases. Cell 103:239–252PubMedCrossRefGoogle Scholar
  6. de Winther MP, van Dijk KW, Havekes LM, Hofker MH (2000) Macrophage scavenger receptor class A: a multifunctional receptor in atherosclerosis. Arterioscler Thromb Vasc Biol 20:290–297PubMedCrossRefGoogle Scholar
  7. Elias A, Tsourkas A (2009) Imaging circulating cells and lymphoid tissues with iron oxide nanoparticles. Hematology Am Soc Hematol Educ Progra 2009:720–726CrossRefGoogle Scholar
  8. Fan F, Jin S, Amundson SA, Tong T, Fan W, Zhao H, Zhu X, Mazzacurati L, Li X, Petrik KL, Fomace AJ Jr, Rajasekaran B, Zhan Z (2002) ATF3 induction following DNA damage is regulated by distinct signaling pathways and over-expression of ATF3 protein suppresses cells growth. Oncogene 21:7488–7496PubMedCrossRefGoogle Scholar
  9. Freeman M, Ashkenas J, Rees DJ, Kingsley DM, Copeland NG, Jenkins NA, Krieger M (1990) An ancient, highly conserved family of cysteine-rich protein domains revealed by cloning type I and type II murine macrophage scavenger receptors. Proc Natl Acad Sci USA 87:8810–8814PubMedCrossRefGoogle Scholar
  10. Gu J, Xu H, Han Y, Dai W, Hao W, Wang C, Gu N, Xu H, Cao J (2011) The internalization pathway, metabolic fate and biological effect of superparamagnetic iron oxide nanoparticles in the macrophage-like RAW264.7 cell. Sci China Life Sci 54:793–805PubMedCrossRefGoogle Scholar
  11. Gupta AK, Gupta M (2005) Synthesis and surface engineering of iron oxide nanoparticles for biomedical applications. Biomaterials 26:3995–4021PubMedCrossRefGoogle Scholar
  12. Hamdi M, Popeijus HE, Carlotti F, Janssen JM, van der Burgt C, Cornelissen-Steijger P, van de Water B, Hoeben RC, Matsuo K, van Dam H (2008) ATF3 and Fra1 have opposite functions in JNK- and ERK-dependent DNA damage responses. DNA Repair (Amst) 7:487–496CrossRefGoogle Scholar
  13. Hussain S, Al-Nsour F, Rice AB, Marshburn J, Yingling B, Ji Z, Zink JI, Walker NJ, Garantziotis S (2012) Cerium dioxide nanoparticles induce apoptosis and autophagy in human peripheral blood monocytes. ACS Nano 6:5820–5829PubMedCrossRefGoogle Scholar
  14. Inoue K, Zama T, Kamimoto T, Aoki R, Ikeda Y, Kimura H, Hagiwara M (2004) TNF alpha-induced ATF3 expression is bidirectionally regulated by the JNK and ERK pathways in vascular endothelial cells. Genes Cells 9:59–70PubMedCrossRefGoogle Scholar
  15. Kajanne R, Miettinen P, Mehlem A, Leivonen SK, Birrer M, Foschi M, Kahari V, Leppa S (2007) EGF-R regulates MMP function in fibroblasts through MAPK and AP-1 pathways. J Cell Physiol 212:489–497PubMedCrossRefGoogle Scholar
  16. Kakiashvili E, Dan Q, Vandermeer M, Zhang Y, Waheed F, Pham M, Szaszi K (2011) The epidermal growth factor receptor mediates tumor necrosis factor-alpha-induced activation of the ERK/GEF-H1/RhoA pathway in tubular epithelium. J Biol Chem 286:9268–9279PubMedCrossRefGoogle Scholar
  17. Kang YS, Sisbud S, Rabolt JF, Stroeve P (1996) Synthesis and characterization of nanometer-size Fe3O4 and γ-Fe2O3 particles. Chem Mater 8:2209CrossRefGoogle Scholar
  18. Kunzmann A, Andersson B, Thumherr T, Krug H, Scheynius A, Fadeel B (2011) Toxicology of engineered nanomaterials: focus on biocompatibility, biodistribution and biodegradation. Biochim Biophys Acta 1810:361–373PubMedCrossRefGoogle Scholar
  19. Laurent S, Boutry S, Mahieu I, Vander Elst L, Muller RN (2009) Iron oxide based MR contrast agents: from chemistry to cell labeling. Curr Med Chem 16:4712–4727PubMedCrossRefGoogle Scholar
  20. Lavicoli I, Leso V, Fontana L, Bergamaschi A (2011) Toxicological effects of titanium dioxide nanoparticles: a review of in vitro mammalian studies. Eur Rev Med Pharmacol Sci 15:481–508Google Scholar
  21. Lee SJ, Pfluger PT, Kim JY, Nogueiras R, Duran A, Pagès G, Pouysségur J, Tschöp MH, Diaz-Meco MT, Moscat J (2010) A functional role for the p62-ERK1 axis in the control of energy homeostasis and adipogenesis. EMBO Rep 11:226–232PubMedCentralPubMedCrossRefGoogle Scholar
  22. Ling D, Hyeon T (2013) Chemical design of biocompatible iron oxide nanoparticles for medical applications. Small 9(9–10):1450–1466Google Scholar
  23. Lopez-Castro JD, Maraloiu AV, Delgado JJ, Calvino JJ, Blanchin MG, Galvez N, Dominguez-Vera JM (2011) From synthetic to natural nanoparticles: monitoring the biodegradation of SPIO (P904) into ferritin by electron microscopy. Nanoscale 3:4597–4599PubMedCrossRefGoogle Scholar
  24. Lu D, Chen J, Hai T (2007) The regulation of ATF3 gene expression by mitogen-activated protein kinases. Biochem J 401:559–567PubMedCrossRefGoogle Scholar
  25. Matsumoto A, Naito M, Itakura H, Ikemoto S, Asaoka H, Hayakawa I, Kanamori H, Aburatani H, Takaku F, Suzuki H, Kobari Y, Miyai T, Takahashi K, Cohenii EH, Wydroii R, Housman DE, Kodama T (1990) Human macrophage scavenger receptors: primary structure, expression, and localization in atherosclerotic lesions. Proc Natl Acad Sci USA 87:9133–9137PubMedCrossRefGoogle Scholar
  26. Naqvi S, Samim M, Abdin M, Ahmed FJ, Maitra A, Prashant C, Dinda AK (2010) Concentration-dependent toxicity of iron oxide nanoparticles mediated by increased oxidative stress. Int J Nanomedicine 5:983–989PubMedCentralPubMedCrossRefGoogle Scholar
  27. Nebreda AR, Porras A (2000) P38 MAP kinases: beyond the stress response. Trends Biochem Sci 25:257–260PubMedCrossRefGoogle Scholar
  28. Nel AE, Madler L, Velegol D, Xia T, Hoek EM, Somasundaran P, Klaessig F, Castranova V, Thompson M (2009) Understanding biophysicochemical interactions at the nano-bio interface. Nat Mater 8:543–557PubMedCrossRefGoogle Scholar
  29. Park EJ, Kim H, Kim Y, Yi J, Choi K, Park K (2010) Inflammatory responses may be induced by a single intratracheal instillation of iron nanoparticles in mice. Toxicology 275:65–71PubMedCrossRefGoogle Scholar
  30. Platt N, Haworth R, Darley L, Gordon S (2002) The many roles of the class A macrophage scavenger receptor. Int Rev Cytol 212:1–40PubMedCrossRefGoogle Scholar
  31. Puppi J, Mitry RR, Modo M, Dhawan A, Raja K, Hughes RD (2011) Use of a clinically approved iron oxide MRI contrast agent to label human hepatocytes. Cell Transplant 20:963–975PubMedCrossRefGoogle Scholar
  32. Roitt IM, Brostoff J, Male DK (1985) Immunology. Gower Medical Pub.; C.V. Mosby, London; New York St. LouisGoogle Scholar
  33. Schrand AM, Rahman MF, Hussain SM, Schlager JJ, Smith DA, Syed AF (2010) Metal-based nanoparticles and their toxicity assessment. Wiley Interdiscip Rev Nanomed Nanobiotechnol 2:544–568PubMedCrossRefGoogle Scholar
  34. Shon HK, Park J, Choi I, Park HM, Moon DW, Lee TG (2011) Mass imaging of iron oxide nanoparticles inside cells for in vitro cytotoxicity. J Nanosci Nanotechnol 11:638–641PubMedCrossRefGoogle Scholar
  35. Simonsen LO, Harbak H, Bennekou P (2012) Cobalt metabolism and toxicology-a brief update. Sci Total Environ 432:210–215PubMedCrossRefGoogle Scholar
  36. Song MM, Song WJ, Bi H, Wang J, Wu WL, Sun J, Yu M (2010) Cytotoxicity and cellular uptake of iron nanowires. Biomaterials 31:1509–1517PubMedCrossRefGoogle Scholar
  37. St Germain C, Niknejad N, Ma L, Garbuio K, Hai T, Dimitroulakos J (2010) Cisplatin induces cytotoxicity through the mitogen-activated protein kinase pathways and activating transcription factor 3. Neoplasia 12:527–538PubMedCentralPubMedGoogle Scholar
  38. Stern ST, Adiseshaiah PP, Crist RM (2012) Autophagy and lysosomal dysfunction as emerging mechanisms of nanomaterial toxicity. Part Fibre Toxicol 14:20CrossRefGoogle Scholar
  39. Subramaniam S, Unsicker K (2006) Extracellular signal-regulated kinase-as an inducer of non-apoptotic neuronal death. Neuroscience 138:1055–1065PubMedCrossRefGoogle Scholar
  40. Suzuki K, Doi T, Imanishi T, Kodama T, Tanaka T (1997a) The conformation of the alpha-helical coiled coil domain of macrophage scavenger receptor is pH dependent. Biochemistry 36:15140–15146PubMedCrossRefGoogle Scholar
  41. Suzuki H, Kurihara Y, Takeya M, Kamada N, Kataoka M, Jishage K, Ueda O, Sakaguchi H, Higashi T, Suzuki T, Takashima Y, Kawabe Y, Cynshi O, Wada Y, Honda M, Kurihara H, Aburatani H, Doi T, Matsumoto A, Azuma S, Noda T, Toyoda Y, Itakura H, Yazaki Y, Horiuchi S, Takahashi K, Kruijt JK, Van Berker T, Steinbrecher UP, Ishibashi S, Maeda N, Gordon S, Kodama T (1997b) A role for macrophage scavenger receptors in atherosclerosis and susceptibility to infection. Nature 386:292–296Google Scholar
  42. Terada Y, Nakashima O, Inoshita S, Kuwahara M, Sasaki S, Marumo F (1999) Mitogen-activated protein kinase cascade and transcription factors: the opposite role of MKK3/6-p38 K and MKK1-MAPK. Nephrol Dial Transplant 14:45–47PubMedCrossRefGoogle Scholar
  43. Wang Y, Wang B, Zhu MT, Li M, Wang HJ, Wang M, Ouyang H, Chai ZF, Feng WY, Zhao YL (2011) Microglial activation, recruitment and phagocytosis as linked phenomena in ferric oxide nanoparticle exposure. Toxicol Lett 205:26–37PubMedCrossRefGoogle Scholar
  44. Yin N, Liu Q, Liu J, He B, Cui L, Li Z, Yun Z, Qu G, Liu S, Zhou Q, Jiang G (2013) Silver nanoparticle exposure attenuates the viability of rat cerebellum granule cells through apoptosis coupled to oxidative stress. Small 9(9–10):1831–1841Google Scholar
  45. Zhao Y, Howe JL, Yu Z, Leong DT, Chu JJ, Loo JS, Ng KW (2013) Exposure to titanium dioxide nanoparticles induces autophagy in primary human keratinocytes. Small 9:387–392PubMedCrossRefGoogle Scholar
  46. Zhu MT, Feng WY, Wang Y, Wang B, Wang M, Ouyang H, Zhao YL, Chai ZF (2009) Particokinetics and extrapulmonary translocation of intratracheally instilled ferric oxide nanoparticles in rats and the potential health risk assessment. Toxicol Sci 107:342–351PubMedCrossRefGoogle Scholar
  47. Zhu XD, Zhuang Y, Ben JJ, Qian LL, Huang HP, Bai H, Sha JH, He ZG, Chen Q (2011) Caveolae-dependent endocytosis is required for class A macrophage scavenger receptor-mediated apoptosis in macrophages. J Biol Chem 286:8231–8239PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.Department of Molecular Science and TechnologyAjou UniversitySuwonKorea
  2. 2.Department of Chemical EngineeringKwangwoon UniversitySeoulKorea
  3. 3.College of Veterinary MedicineSeoul National UniversitySeoulKorea
  4. 4.Department of Nano Fusion Technology, Graduate School of Convergence Science and TechnologySeoul National UniversitySuwonKorea
  5. 5.Graduate Group of Tumor BiologySeoul National UniversitySeoulKorea
  6. 6.Advanced Institute of Convergence TechnologySeoul National UniversitySuwonKorea

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