Skip to main content
SpringerLink
Log in

Systemic Lipopolysaccharide Induces Cochlear Inflammation and Exacerbates the Synergistic Ototoxicity of Kanamycin and Furosemide

  • Research Article
  • Published:
Journal of the Association for Research in Otolaryngology Aims and scope Submit manuscript

Abstract

Aminoglycoside antibiotics are highly effective agents against gram-negative bacterial infections, but they cause adverse effects on hearing and balance dysfunction as a result of toxicity to hair cells of the cochlea and vestibular organs. While ototoxicity has been comprehensively studied, the contributions of the immune system, which controls the host response to infection, have not been studied in antibiotic ototoxicity. Recently, it has been shown that an inflammatory response is induced by hair cell injury. In this study, we found that lipopolysaccharide (LPS), an important component of bacterial endotoxin, when given in combination with kanamycin and furosemide, augmented the inflammatory response to hair cell injury and exacerbated hearing loss and hair cell injury. LPS injected into the peritoneum of experimental mice induced a brisk cochlear inflammatory response with recruitment of mononuclear phagocytes into the spiral ligament, even in the absence of ototoxic agents. While LPS alone did not affect hearing, animals that received LPS prior to ototoxic agents had worse hearing loss compared to those that did not receive LPS pretreatment. The poorer hearing outcome in LPS-treated mice did not correlate to changes in endocochlear potential. However, LPS-treated mice demonstrated an increased number of CCR2+ inflammatory monocytes in the inner ear when compared with mice treated with ototoxic agents alone. We conclude that LPS and its associated inflammatory response are harmful to the inner ear when coupled with ototoxic medications and that the immune system may contribute to the final hearing outcome in subjects treated with ototoxic agents.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

FIG. 1
FIG. 2
FIG. 3
FIG. 4
FIG. 5
FIG. 6
FIG. 7
FIG. 8

Similar content being viewed by others

References

  • Arnold W, Nadol JB Jr, Weidauer H (1981) Ultrastructural histopathology in a case of human ototoxicity due to loop diuretics. Acta Otolaryngol 91(5–6):399–414

    Article  CAS  PubMed  Google Scholar 

  • Asakuma S, Snow JB Jr (1980) Effects of furosemide and ethacrynic acid on the endocochlear direct current potential in normal and kanamycin sulfate-treated guinea pigs. Otolaryngol Head Neck Surg 88(2):188–193

    CAS  PubMed  Google Scholar 

  • Auffray C, Fogg D, Garfa M, Elain G, Join-Lambert O, Kayal S, Sarnacki S, Cumano A, Lauvau G, Geissmann F (2007) Monitoring of blood vessels and tissues by a population of monocytes with patrolling behavior. Science 317(5838):666–670. doi:10.1126/science.1142883

    Article  CAS  PubMed  Google Scholar 

  • Azuma H, Takeuchi S, Higashiyama K, Ando M, Kakigi A, Nakahira M, Yamakawa K, Takeda T (2002) Bumetanide-induced enlargement of the intercellular space in the stria vascularis requires an active Na + -K + -ATPase. Acta Otolaryngol 122(8):816–821

    Article  CAS  PubMed  Google Scholar 

  • Bauerfeld CP, Rastogi R, Pirockinaite G, Lee I, Huttemann M, Monks B, Birnbaum MJ, Franchi L, Nunez G, Samavati L (2012) TLR4-mediated AKT activation is MyD88/TRIF dependent and critical for induction of oxidative phosphorylation and mitochondrial transcription factor A in murine macrophages. J Immunol 188(6):2847–2857. doi:10.4049/jimmunol.1102157

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bosmann M, Russkamp NF, Ward PA (2012) Fingerprinting of the TLR4-induced acute inflammatory response. Exp Mol Pathol 93(3):319–323. doi:10.1016/j.yexmp.2012.08.006

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Chakravarty S, Herkenham M (2005) Toll-like receptor 4 on nonhematopoietic cells sustains CNS inflammation during endotoxemia, independent of systemic cytokines. J Neurosci Off J Soc Neurosci 25(7):1788–1796. doi:10.1523/JNEUROSCI.4268-04.2005

    Article  CAS  Google Scholar 

  • De Arras L, Seng A, Lackford B, Keikhaee MR, Bowerman B, Freedman JH, Schwartz DA, Alper S (2013) An evolutionarily conserved innate immunity protein interaction network. J Biol Chem 288(3):1967–1978. doi:10.1074/jbc.M112.407205

    Article  PubMed Central  PubMed  Google Scholar 

  • Deng JC, Cheng G, Newstead MW, Zeng X, Kobayashi K, Flavell RA, Standiford TJ (2006) Sepsis-induced suppression of lung innate immunity is mediated by IRAK-M. J Clin Invest 116(9):2532–2542. doi:10.1172/JCI28054

    CAS  PubMed Central  PubMed  Google Scholar 

  • Ding D, McFadden SL, Woo JM, Salvi RJ (2002) Ethacrynic acid rapidly and selectively abolishes blood flow in vessels supplying the lateral wall of the cochlea. Hear Res 173(1–2):1–9

    Article  CAS  PubMed  Google Scholar 

  • Ehret G (1983) Peripheral anatomy and physiology II. In: Willott J (ed) The auditory psychobiology of the mouse. Charles C. Thomas, Springfield, pp 169–200

    Google Scholar 

  • Figueroa L, Xiong Y, Song C, Piao W, Vogel SN, Medvedev AE (2012) The Asp299Gly polymorphism alters TLR4 signaling by interfering with recruitment of MyD88 and TRIF. J Immunol 188(9):4506–4515. doi:10.4049/jimmunol.1200202

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Fredelius L, Rask-Andersen H (1990) The role of macrophages in the disposal of degeneration products within the organ of corti after acoustic overstimulation. Acta Otolaryngol 109(1–2):76–82

    Article  CAS  PubMed  Google Scholar 

  • Geissmann F, Jung S, Littman DR (2003) Blood monocytes consist of two principal subsets with distinct migratory properties. Immunity 19(1):71–82

    Article  CAS  PubMed  Google Scholar 

  • Guo Y, Wu Y, Chen W, Lin J (1994) Endotoxic damage to the stria vascularis: the pathogenesis of sensorineural hearing loss secondary to otitis media? J Laryngol Otol 108(4):310–313

    Article  CAS  PubMed  Google Scholar 

  • Hequembourg S, Liberman MC (2001) Spiral ligament pathology: a major aspect of age-related cochlear degeneration in C57BL/6 mice. J Assoc Res Otolaryngol: JARO 2(2):118–129

    CAS  PubMed Central  PubMed  Google Scholar 

  • Hirose K, Sato E (2011) Comparative analysis of combination kanamycin-furosemide versus kanamycin alone in the mouse cochlea. Hear Res 272(1–2):108–116. doi:10.1016/j.heares.2010.10.011

    Article  CAS  PubMed  Google Scholar 

  • Hirose K, Discolo CM, Keasler JR, Ransohoff R (2005) Mononuclear phagocytes migrate into the murine cochlea after acoustic trauma. J Comp Neurol 489(2):180–194

    Article  PubMed  Google Scholar 

  • Jangula A, Murphy EJ (2013) Lipopolysaccharide-induced blood brain barrier permeability is enhanced by alpha-synuclein expression. Neurosci Lett 551:23–27. doi:10.1016/j.neulet.2013.06.058

    Article  CAS  PubMed  Google Scholar 

  • Jung S, Aliberti J, Graemmel P, Sunshine MJ, Kreutzberg GW, Sher A, Littman DR (2000) Analysis of fractalkine receptor CX(3)CR1 function by targeted deletion and green fluorescent protein reporter gene insertion. Mol Cell Biol 20(11):4106–4114

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kang J, Rivest S (2007) MyD88-deficient bone marrow cells accelerate onset and reduce survival in a mouse model of amyotrophic lateral sclerosis. J Cell Biol 179(6):1219–1230. doi:10.1083/jcb.200705046

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kim CS, Kim HJ (1995) Auditory brain stem response changes after application of endotoxin to the round window membrane in experimental otitis media. Otolaryngol Head Neck Surg: Off J Am Acad Otolaryngol Head Neck Surg 112(4):557–565

    Article  CAS  Google Scholar 

  • Komune S, Snow JB Jr (1982) Nature of the endocochlear dc potential in kanamycin-poisoned guinea pigs. Arch Otolaryngol 108(6):334–338

    Article  CAS  PubMed  Google Scholar 

  • Lang H, Schulte BA, Schmiedt RA (2003) Effects of chronic furosemide treatment and age on cell division in the adult gerbil inner ear. J Assoc Res Otolaryngol: JARO 4(2):164–175

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lustig S, Danenberg HD, Kafri Y, Kobiler D, Ben-Nathan D (1992) Viral neuroinvasion and encephalitis induced by lipopolysaccharide and its mediators. J Exp Med 176(3):707–712

    Article  CAS  PubMed  Google Scholar 

  • Marsh B, Stevens SL, Packard AE, Gopalan B, Hunter B, Leung PY, Harrington CA, Stenzel-Poore MP (2009) Systemic lipopolysaccharide protects the brain from ischemic injury by reprogramming the response of the brain to stroke: a critical role for IRF3. J Neurosci Off J Soc Neurosci 29(31):9839–9849. doi:10.1523/JNEUROSCI.2496-09.2009

    Article  CAS  Google Scholar 

  • Mathog RH, Thomas WG, Hudson WR (1970) Ototoxicity of new and potent diuretics. A preliminary study. Arch Otolaryngol 92(1):7–13

    Article  CAS  PubMed  Google Scholar 

  • Maus UA, Koay MA, Delbeck T, Mack M, Ermert M, Ermert L, Blackwell TS, Christman JW, Schlondorff D, Seeger W, Lohmeyer J (2002) Role of resident alveolar macrophages in leukocyte traffic into the alveolar air space of intact mice. Am J Physiol Lung Cell Mol Physiol 282(6):L1245–L1252. doi:10.1152/ajplung.00453.2001

    CAS  PubMed  Google Scholar 

  • Nadeau S, Rivest S (2002) Endotoxemia prevents the cerebral inflammatory wave induced by intraparenchymal lipopolysaccharide injection: role of glucocorticoids and CD14. J Immunol 169(6):3370–3381

    Article  CAS  PubMed  Google Scholar 

  • Nahid MA, Satoh M, Chan EK (2011) MicroRNA in TLR signaling and endotoxin tolerance. Cell Mol Immunol 8(5):388–403. doi:10.1038/cmi.2011.26

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Naito H, Watanabe K (1997) Alteration in capillary permeability of horseradish peroxidase in the stria vascularis and movement of leaked horseradish peroxidase after administration of furosemide. ORL J Otorhinolaryngol Relat Spec 59(5):248–257

    Article  CAS  PubMed  Google Scholar 

  • Nishioku T, Dohgu S, Takata F, Eto T, Ishikawa N, Kodama KB, Nakagawa S, Yamauchi A, Kataoka Y (2009) Detachment of brain pericytes from the basal lamina is involved in disruption of the blood-brain barrier caused by lipopolysaccharide-induced sepsis in mice. Cell Mol Neurobiol 29(3):309–316. doi:10.1007/s10571-008-9322-x

    Article  CAS  PubMed  Google Scholar 

  • O'Dea KP, Wilson MR, Dokpesi JO, Wakabayashi K, Tatton L, van Rooijen N, Takata M (2009) Mobilization and margination of bone marrow Gr-1high monocytes during subclinical endotoxemia predisposes the lungs toward acute injury. J Immunol 182(2):1155–1166

    Article  PubMed Central  PubMed  Google Scholar 

  • Oesterle EC, Campbell S (2009) Supporting cell characteristics in long-deafened aged mouse ears. J Assoc Res Otolaryngol: JARO 10(4):525–544. doi:10.1007/s10162-009-0183-x

    Article  PubMed Central  PubMed  Google Scholar 

  • Oesterle EC, Campbell S, Taylor RR, Forge A, Hume CR (2008) Sox2 and JAGGED1 expression in normal and drug-damaged adult mouse inner ear. J Assoc Res Otolaryngol: JARO 9(1):65–89. doi:10.1007/s10162-007-0106-7

    Article  PubMed Central  PubMed  Google Scholar 

  • Oh GS, Kim HJ, Choi JH, Shen A, Kim CH, Kim SJ, Shin SR, Hong SH, Kim Y, Park C, Lee SJ, Akira S, Park R, So HS (2011) Activation of lipopolysaccharide-TLR4 signaling accelerates the ototoxic potential of cisplatin in mice. J Immunol 186(2):1140–1150. doi:10.4049/jimmunol.1002183

    Article  CAS  PubMed  Google Scholar 

  • Ohlemiller KK (2009) Mechanisms and genes in human strial presbycusis from animal models. Brain Res 1277:70–83. doi:10.1016/j.brainres.2009.02.079

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Quintanilla-Dieck L, Larrain B, Trune D, Steyger PS (2013) Effect of systemic lipopolysaccharide-induced inflammation on cytokine levels in the murine cochlea: a pilot study. Otolaryngol Head Neck Surg: Off J Am Acad Otolaryngol Head Neck Surg 149(2):301–303. doi:10.1177/0194599813491712

    Article  Google Scholar 

  • Rosenzweig HL, Lessov NS, Henshall DC, Minami M, Simon RP, Stenzel-Poore MP (2004) Endotoxin preconditioning prevents cellular inflammatory response during ischemic neuroprotection in mice. Stroke 35(11):2576–2581

    Article  CAS  PubMed  Google Scholar 

  • Rybak LP (1993) Ototoxicity of loop diuretics. Otolaryngol Clin N Am 26(5):829–844

    CAS  Google Scholar 

  • Rybak LP, Whitworth C, Scott V (1991) Comparative acute ototoxicity of loop diuretic compounds. Eur Arch Otorhinolaryngol 248(6):353–357

    CAS  PubMed  Google Scholar 

  • Saederup N, Cardona AE, Croft K, Mizutani M, Cotleur AC, Tsou CL, Ransohoff RM, Charo IF (2010) Selective chemokine receptor usage by central nervous system myeloid cells in CCR2-red fluorescent protein knock-in mice. PLoS One 5(10):e13693. doi:10.1371/journal.pone.0013693

    Article  PubMed Central  PubMed  Google Scholar 

  • Saha P, Geissmann F (2011) Toward a functional characterization of blood monocytes. Immunol Cell Biol 89(1):2–4. doi:10.1038/icb.2010.130

    Article  PubMed  Google Scholar 

  • Sato E, Shick HE, Ransohoff RM, Hirose K (2008) Repopulation of cochlear macrophages in murine hematopoietic progenitor cell chimeras: the role of CX3CR1. J Comp Neurol 506(6):930–942

    Article  PubMed  Google Scholar 

  • Sato E, Shick HE, Ransohoff RM, Hirose K (2010) Expression of fractalkine receptor CX3CR1 on cochlear macrophages influences survival of hair cells following ototoxic injury. J Assoc Res Otolaryngol: JARO 11(2):223–234. doi:10.1007/s10162-009-0198-3

    Article  PubMed Central  PubMed  Google Scholar 

  • Schulz C, Gomez Perdiguero E, Chorro L, Szabo-Rogers H, Cagnard N, Kierdorf K, Prinz M, Wu B, Jacobsen SE, Pollard JW, Frampton J, Liu KJ, Geissmann F (2012) A lineage of myeloid cells independent of Myb and hematopoietic stem cells. Science 336(6077):86–90. doi:10.1126/science.1219179

    Article  CAS  PubMed  Google Scholar 

  • Sewell WF (1984) Furosemide selectively reduces one component in rate-level functions from auditory-nerve fibers. Hear Res 15(1):69–72

    Article  CAS  PubMed  Google Scholar 

  • Shi J, Johansson J, Woodling NS, Wang Q, Montine TJ, Andreasson K (2010) The prostaglandin E2 E-prostanoid 4 receptor exerts anti-inflammatory effects in brain innate immunity. J Immunol 184(12):7207–7218. doi:10.4049/jimmunol.0903487

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Simard AR, Rivest S (2007) Neuroprotective effects of resident microglia following acute brain injury. J Comp Neurol 504(6):716–729. doi:10.1002/cne.21469

    Article  CAS  PubMed  Google Scholar 

  • Tan BT, Lee MM, Ruan R (2008) Bone-marrow-derived cells that home to acoustic deafened cochlea preserved their hematopoietic identity. J Comp Neurol 509(2):167–179. doi:10.1002/cne.21729

    Article  PubMed  Google Scholar 

  • Tauseef M, Knezevic N, Chava KR, Smith M, Sukriti S, Gianaris N, Obukhov AG, Vogel SM, Schraufnagel DE, Dietrich A, Birnbaumer L, Malik AB, Mehta D (2012) TLR4 activation of TRPC6-dependent calcium signaling mediates endotoxin-induced lung vascular permeability and inflammation. J Exp Med 209(11):1953–1968. doi:10.1084/jem.20111355

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Taylor RR, Nevill G, Forge A (2008) Rapid hair cell loss: a mouse model for cochlear lesions. J Assoc Res Otolaryngol: JARO 9(1):44–64

    Article  PubMed Central  PubMed  Google Scholar 

  • Tornabene SV, Sato K, Pham L, Billings P, Keithley EM (2006) Immune cell recruitment following acoustic trauma. Hear Res 222(1-2):115–124. doi:10.1016/j.heares.2006.09.004

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Hirose K, Liberman MC (2002) Dynamics of noise-induced cellular injury and repair in the mouse cochlea. J Assoc Res Otolaryngol: JARO 3(3):248–268

    Article  PubMed Central  PubMed  Google Scholar 

  • Wang W, Deng M, Liu X, Ai W, Tang Q, Hu J (2011) TLR4 activation induces nontolerant inflammatory response in endothelial cells. Inflammation 34(6):509–518. doi:10.1007/s10753-010-9258-4

    Article  CAS  PubMed  Google Scholar 

  • Warchol ME (1997) Macrophage activity in organ cultures of the avian cochlea: demonstration of a resident population and recruitment to sites of hair cell lesions. J Neurobiol 33(6):724–734. doi:10.1002/(SICI)1097-4695(19971120)33:6<724::AID-NEU2>3.0.CO;2-B

    Article  CAS  PubMed  Google Scholar 

  • Wispelwey B, Lesse AJ, Hansen EJ, Scheld WM (1988) Haemophilus influenzae lipopolysaccharide-induced blood brain barrier permeability during experimental meningitis in the rat. J Clin Invest 82(4):1339–1346. doi:10.1172/JCI113736

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wu WJ, Sha SH, McLaren JD, Kawamoto K, Raphael Y, Schacht J (2001) Aminoglycoside ototoxicity in adult CBA, C57BL and BALB mice and the Sprague-Dawley rat. Hear Res 158(1–2):165–178

    Article  CAS  PubMed  Google Scholar 

  • Xiong Y, Medvedev AE (2011) Induction of endotoxin tolerance in vivo inhibits activation of IRAK4 and increases negative regulators IRAK-M, SHIP-1, and A20. J Leukoc Biol 90(6):1141–1148. doi:10.1189/jlb.0611273

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

Many thanks to Drs. Mark Warchol and Alec Salt for careful feedback on the manuscript and to Dorina Kallojieri for assistance with statistical analysis. Funding sources supporting this work include NIH DC011315 and a research grant from the American Otological Society.

Conflict of Interest

None of the authors who have authored or provided materials for this work have a financial, personal, or other conflicting interest in the results of this research or publication of this work.

Author Contributions

All authors had full access to the data in this study and take responsibility for the integrity and accuracy of the data analysis. Study concept and design: KH, KKO, RMR. Acquisition of data: SZL, KKO, KH. Analysis and interpretations of the data: KH, KKO, RMR. Writing of the manuscript: KH. Statistical analysis: KH. Funding awarded to: KH. Study supervision: KH.

Author information

Authors and Affiliations

  1. Department of Otolaryngology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO, 63110, USA

    Keiko Hirose, Song-Zhe Li & Kevin K. Ohlemiller

  2. Neuroinflammation Research Center, Department of Neurosciences, Cleveland Clinic Lerner Research Institute, Cleveland, OH, 44195, USA

    Richard M. Ransohoff

Authors
  1. Keiko Hirose
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Song-Zhe Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kevin K. Ohlemiller
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Richard M. Ransohoff
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Keiko Hirose.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hirose, K., Li, SZ., Ohlemiller, K.K. et al. Systemic Lipopolysaccharide Induces Cochlear Inflammation and Exacerbates the Synergistic Ototoxicity of Kanamycin and Furosemide. JARO 15, 555–570 (2014). https://doi.org/10.1007/s10162-014-0458-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10162-014-0458-8

Keywords

Search

Navigation