Autophagy upregulation promotes macrophages to escape mesoporous silica nanoparticle (MSN)-induced NF-κB-dependent inflammation

BackgroundOur previous studies (Int J Nanomed 10:22, 2015) have indicated that a single large dose of mesoporous silica nanoparticles (MSNs) can induce severe and selective nephrotoxicity, which is closely related to inflammation mediated by the NF-κB pathway. However, the effect of MSNs on other organs and the interactions of nanomaterials with biological systems remain rudimentary.ObjectiveThis study aimed to clarify the biological behaviour and influence of MSNs on macrophages.MethodsThe mice received a single intraperitoneal injection of a suspension of 150, 300 of 600 mg/kg MSNs, and RAW 264.7 cells were treated with MSNs at various concentrations and times. Cell viability was determined by MTT assay and LDH release assay. The NF-κB pathway and the target proinflammatory cytokines IL-1β and TNF-α were determined by western blotting or ELISA. Autophagy is considered as an emerging mechanism of nanomaterials. So the autophagic ultrastructural analysis, the determination of Beclin-1 and LC3 expression, and the calculation of LC3II dots were employed to verify autophagy activation. In addition, RNA interference, autophagy agonist and inhibitor were used to explore the role of autophagy in inflammation.ResultsThe results indicated that MSNs are internalized into macrophages and induce cytotoxicity in a dose- and time-dependent manner. The NF-κB pathway, IL-1β and TNF-α were induced and released by MSNs. The levels of Beclin-1 and LC3II dots were obviously up-regulated by MSNs, which indicated that autophagy was induced in the MSN-treated cells. Moreover, the enhanced autophagy can attenuate the inflammation mediated by the NF-κB pathway, whereas the inhibition of autophagy can contribute to inflammation.ConclusionsIn summary, our results suggest that autophagy may be a possible protective factor in inflammation induced by MSNs in macrophages.

[1]  Suthida Boonsith,et al.  The Practicality of Mesoporous Silica Nanoparticles as Drug Delivery Devices and Progress Toward This Goal , 2014, AAPS PharmSciTech.

[2]  Y. Tsutsumi,et al.  Histological analysis of 70-nm silica particles-induced chronic toxicity in mice. , 2009, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[3]  Toshiro Hirai,et al.  Amorphous nanosilicas induce consumptive coagulopathy after systemic exposure , 2012, Nanotechnology.

[4]  Dong Chen,et al.  Pathological mechanisms of liver injury caused by continuous intraperitoneal injection of silica nanoparticles. , 2012, Biomaterials.

[5]  V. S. Lin,et al.  Endocytosis of a single mesoporous silica nanoparticle into a human lung cancer cell observed by differential interference contrast microscopy , 2008, Analytical and bioanalytical chemistry.

[6]  J. Kang,et al.  Src tyrosine kinases mediate crystalline silica-induced NF-kappaB activation through tyrosine phosphorylation of IkappaB-alpha and p65 NF-kappaB in RAW 264.7 macrophages. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[7]  T. Hartung,et al.  Autophagy as an ultrastructural marker of heavy metal toxicity in human cord blood hematopoietic stem cells. , 2008, The Science of the total environment.

[8]  G. Milazzo.  IUPAC - Manual of Symbols and Terminology for Physico-chemical Quantities and Units. : D.H. Whiffen (Editor). Pergamon Press, Oxford, 1979 ed., 41 pp. , 1981 .

[9]  Jeffrey I Zink,et al.  Functional nanovalves on protein-coated nanoparticles for in vitro and in vivo controlled drug delivery. , 2015, Small.

[10]  M. Zern,et al.  A role for tissue transglutaminase in hepatic injury and fibrogenesis, and its regulation by NF-kappaB. , 1997, The American journal of physiology.

[11]  Soyoung Lee,et al.  The comparative effects of mesoporous silica nanoparticles and colloidal silica on inflammation and apoptosis. , 2011, Biomaterials.

[12]  Z. Kmieć,et al.  Introduction — Morphology of the Liver Lobule , 2001 .

[13]  Y. Li,et al.  Silica nanoparticles induce autophagy and endothelial dysfunction via the PI3K/Akt/mTOR signaling pathway , 2014, International journal of nanomedicine.

[14]  D. Sabatini,et al.  mTOR: from growth signal integration to cancer, diabetes and ageing , 2010, Nature Reviews Molecular Cell Biology.

[15]  N. Gu,et al.  Inhibition of autophagy enhances the anticancer activity of silver nanoparticles , 2014, Autophagy.

[16]  Meiying Wang,et al.  Interference in Autophagosome Fusion by Rare Earth Nanoparticles Disrupts Autophagic Flux and Regulation of an Interleukin-1β Producing Inflammasome , 2014, ACS nano.

[17]  H. Tsutsui,et al.  Importance of Kupffer Cells in the Development of Acute Liver Injuries in Mice , 2014, International journal of molecular sciences.

[18]  T. Pan,et al.  Rapamycin protects against rotenone-induced apoptosis through autophagy induction , 2009, Neuroscience.

[19]  Jae-Ho Kim,et al.  Magnetic iron oxide nanoparticles induce autophagy preceding apoptosis through mitochondrial damage and ER stress in RAW264.7 cells. , 2014, Toxicology in vitro : an international journal published in association with BIBRA.

[20]  Keyvan Nasirzadeh,et al.  In vivo toxicity of orally administrated silicon dioxide nanoparticles in healthy adult mice , 2014, Environmental Science and Pollution Research.

[21]  Juan L. Vivero-Escoto,et al.  Mesoporous silica nanoparticles for reducing hemolytic activity towards mammalian red blood cells. , 2009, Small.

[22]  Markus R. Wenk,et al.  Dual Role of 3-Methyladenine in Modulation of Autophagy via Different Temporal Patterns of Inhibition on Class I and III Phosphoinositide 3-Kinase* , 2010, The Journal of Biological Chemistry.

[23]  Shuming Nie,et al.  Bioconjugated quantum dots for in vivo molecular and cellular imaging. , 2008, Advanced drug delivery reviews.

[24]  Wen-Qi Jiang,et al.  Autophagy-related proteins Beclin-1 and LC3 predict cetuximab efficacy in advanced colorectal cancer. , 2011, World journal of gastroenterology.

[25]  W. Duan,et al.  Graphene quantum dots induce apoptosis, autophagy, and inflammatory response via p38 mitogen-activated protein kinase and nuclear factor-κB mediated signaling pathways in activated THP-1 macrophages. , 2015, Toxicology.

[26]  J. Hoek,et al.  Ethanol, oxidative stress, and cytokine-induced liver cell injury. , 2002, Alcohol.

[27]  N. Mizushima,et al.  Methods in Mammalian Autophagy Research , 2010, Cell.

[28]  D. H. Everett,et al.  Manual of Symbols and Terminology for Physicochemical Quantities and Units, Appendix II: Definitions, Terminology and Symbols in Colloid and Surface Chemistry , 1972 .

[29]  I. Tanida.  Autophagosome formation and molecular mechanism of autophagy. , 2011, Antioxidants & redox signaling.

[30]  S. K. Li,et al.  Silicone Adhesive Matrix of Verapamil Hydrochloride to Provide pH-Independent Sustained Release , 2014, AAPS PharmSciTech.

[31]  Xinglu Huang,et al.  Single and repeated dose toxicity of mesoporous hollow silica nanoparticles in intravenously exposed mice. , 2011, Biomaterials.

[32]  Y. Doki,et al.  LC3, an autophagosome marker, is highly expressed in gastrointestinal cancers. , 2008, International journal of oncology.

[33]  Juan L. Vivero-Escoto,et al.  Mesoporous silica nanoparticles as controlled release drug delivery and gene transfection carriers. , 2008, Advanced drug delivery reviews.

[34]  S. Ryter,et al.  Autophagy: A critical regulator of cellular metabolism and homeostasis , 2013, Molecules and cells.

[35]  M. Ratain,et al.  Rapamycin: Something Old, Something New, Sometimes Borrowed and Now Renewed , 2007, Clinical pharmacology and therapeutics.

[36]  M. Muders,et al.  Autophagy: Detection, regulation and its role in cancer and therapy response , 2014, International journal of radiation biology.

[37]  J. Kang,et al.  Src tyrosine kinases mediate crystalline silica-induced NF-κB activation through tyrosine phosphorylation of IκB-α and p65 NF-κB in raw 264.7 macrophages , 2006 .

[38]  Dong Chen,et al.  Multifunctional gold nanoshells on silica nanorattles: a platform for the combination of photothermal therapy and chemotherapy with low systemic toxicity. , 2011, Angewandte Chemie.

[39]  S. Garantziotis,et al.  Interplay between apoptotic and autophagy pathways after exposure to cerium dioxide nanoparticles in human monocytes , 2013, Autophagy.

[40]  Yuan-Lu Cui,et al.  Dietary Blue Pigments Derived from Genipin, Attenuate Inflammation by Inhibiting LPS-Induced iNOS and COX-2 Expression via the NF-κB Inactivation , 2012, PloS one.

[41]  M. Saboungi,et al.  Mesoporous silica nanoparticles enhance MTT formazan exocytosis in HeLa cells and astrocytes. , 2009, Toxicology in vitro : an international journal published in association with BIBRA.

[42]  R. Teng,et al.  Cross talk between NADPH oxidase and autophagy in pulmonary artery endothelial cells with intrauterine persistent pulmonary hypertension. , 2012, American journal of physiology. Lung cellular and molecular physiology.

[43]  J. Choy,et al.  Human-related application and nanotoxicology of inorganic particles: complementary aspects , 2008 .

[44]  Yang Yu,et al.  Silica nanoparticles enhance autophagic activity, disturb endothelial cell homeostasis and impair angiogenesis , 2014, Particle and Fibre Toxicology.

[45]  H. Ghandehari,et al.  Silica nanoconstruct cellular toleration threshold in vitro. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[46]  Z. Kmieć,et al.  Cooperation of Liver Cells in Health and Disease , 2001, Advances in Anatomy Embryology and Cell Biology.

[47]  Linlin Li,et al.  The absorption, distribution, excretion and toxicity of mesoporous silica nanoparticles in mice following different exposure routes. , 2013, Biomaterials.

[48]  Y. Yoshioka,et al.  Systemic distribution, nuclear entry and cytotoxicity of amorphous nanosilica following topical application. , 2011, Biomaterials.

[49]  X. Chen,et al.  Renal interstitial fibrosis induced by high-dose mesoporous silica nanoparticles via the NF-κB signaling pathway , 2014, International journal of nanomedicine.

[50]  N. Grishin,et al.  EGFR-Mediated Beclin 1 Phosphorylation in Autophagy Suppression, Tumor Progression, and Tumor Chemoresistance , 2013, Cell.

[51]  M. Yezhelyev,et al.  Nanoparticles as a Novel Class of Autophagy Activators , 2007, Autophagy.

[52]  Rachael M. Crist,et al.  Autophagy and lysosomal dysfunction as emerging mechanisms of nanomaterial toxicity , 2012, Particle and Fibre Toxicology.

[53]  F. Rossi,et al.  Silica nanoparticle uptake induces survival mechanism in A549 cells by the activation of autophagy but not apoptosis. , 2014, Toxicology letters.

[54]  T. Noda,et al.  Monitoring autophagy in mammalian cultured cells through the dynamics of LC3. , 2009, Methods in enzymology.

[55]  Tingting Ding,et al.  SiO2 nanoparticle-induced impairment of mitochondrial energy metabolism in hepatocytes directly and through a Kupffer cell-mediated pathway in vitro , 2014, International journal of nanomedicine.

[56]  J. Gong,et al.  An efficient method to isolate and culture mouse Kupffer cells. , 2014, Immunology letters.

[57]  Qianjun He,et al.  MSN‐Mediated Sequential Vascular‐to‐Cell Nuclear‐Targeted Drug Delivery for Efficient Tumor Regression , 2014, Advanced materials.

[58]  G. Oberdörster,et al.  Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles , 2005, Environmental health perspectives.

[59]  T. Asefa,et al.  Mesoporosity and functional group dependent endocytosis and cytotoxicity of silica nanomaterials. , 2009, Chemical research in toxicology.

[60]  W. Chan,et al.  Nanotoxicity: the growing need for in vivo study. , 2007, Current opinion in biotechnology.

[61]  Takeshi Noda,et al.  LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing , 2000, The EMBO journal.

[62]  Ruchi Roy,et al.  Zinc oxide nanoparticles induce apoptosis by enhancement of autophagy via PI3K/Akt/mTOR inhibition. , 2014, Toxicology letters.

[63]  T. Asefa,et al.  Mesoporous silica nanoparticles inhibit cellular respiration. , 2008, Nano letters.

[64]  D. Tang,et al.  Chloroquine inhibits HMGB1 inflammatory signaling and protects mice from lethal sepsis. , 2013, Biochemical pharmacology.

[65]  Wei Xu,et al.  Methyl-1-hydroxy-2-naphthoate, a novel naphthol derivative, inhibits lipopolysaccharide-induced inflammatory response in macrophages via suppression of NF-κB, JNK and p38 MAPK pathways , 2011, Inflammation Research.

[66]  Liang Jinqiang,et al.  Triptolide-induced oxidative stress involved with Nrf2 contribute to cardiomyocyte apoptosis through mitochondrial dependent pathways. , 2014, Toxicology letters.

[67]  Christian Mühlfeld,et al.  Translocation and cellular entering mechanisms of nanoparticles in the respiratory tract. , 2008, Swiss medical weekly.