Clin Exp Otorhinolaryngol.  2015 Jun;8(2):123-128. 10.3342/ceo.2015.8.2.123.

Increase in the Level of Proinflammatory Cytokine HMGB1 in Nasal Fluids of Patients With Rhinitis and its Sequestration by Glycyrrhizin Induces Eosinophil Cell Death

• Cavone L
• Cuppari C
• Manti S
• Grasso L
• Arrigo T
• Calamai L
• Salpietro
• Chiarugi A

• KMID: 2360782
• DOI: http://doi.org/10.3342/ceo.2015.8.2.123

Abstract

OBJECTIVES
The nuclear protein high mobility group protein box 1 (HMGB1) is a proinflammatory mediator that belongs to the alarmin family of proinflammatory mediators, and it has recently emerged as a key player in different acute and chronic immune disorders. Several lines of evidence demonstrate that HMGB1 is actively released extracellularly from immune cells or passively released from necrotic cells. Because of the ability of HMGB1 to sustain chronic inflammation, we investigated whether the protein is present in nasal fluids of patients with different forms of rhinitis.
METHODS
HMGB1 levels were evaluated in nasal fluids of healthy subjects or rhinitis patients who were treated or not treated with different treatments.
RESULTS
We report that the level of HMGB1 was significantly increased in nasal fluids of patients with allergic rhinitis, patients with NARES (nonallergic rhinitis with eosinophiliac syndrome), as well as patients with polyps. We also found that a formulation containing the HMGB1-binding compound glycyrrhizin (GLT) reduced the HMGB1 content in nasal fluids of rhinitis patients to an extent similar to that with nasal budesonide treatment. We also found that among the cultured human leukocyte populations, eosinophils released higher amounts of HMGB1. Based on the ability of HMGB1 to sustain eosinophil survival and the ability of GLT to inactivate HMGB1, we report that GLT selectively killed cultured eosinophils and had no effect on neutrophils, macrophages, and lymphocytes.
CONCLUSION
Collectively, these data underscore the role of HMGB1 in rhinitis pathogenesis and the therapeutic potential of GLT formulations in treatment of chronic inflammatory disorders of the nasal mucosa.

Keyword

Rhinitis; Eosinophils; HMGB1 protein; Glycyrrhizin acid

MeSH Terms

Budesonide
Cell Death*
Eosinophils*
Glycyrrhizic Acid*
HMGB1 Protein*
Humans
Immune System Diseases
Inflammation
Leukocytes
Lymphocytes
Macrophages
Nasal Mucosa
Neutrophils
Nuclear Proteins
Polyps
Rhinitis*
Budesonide
Glycyrrhizic Acid
HMGB1 Protein
Nuclear Proteins

Figure

• 

  Fig. 1 High mobility group protein box 1 (HMGB1) levels in rhinitis patients, plasma levels of glycyrrhizin, glycyrrhetinic acid (GA), and cortisol in patients treated with Narivent (DMG, Rome, Italy) or GLT-containing formulation. (A) HMGB1 content in nasal fluids of healthy subjects and patients with different forms of rhinitis/polyps. (B) Comparison of the HMGB1 content in nasal fluids of healthy subjects or subjects affected by rhinitis. (C) HMGB1 levels in nasal fluids of patients with allergic rhinitis before and after 1 week of treatment with saline, Narivent, or Budesonide. (D) Levels of GLT and GA in subjects treated with acute (i.e., 30 minutes after 1 puff/nostril) or chronic (1 puff/nostril twice a day for 1 week) Narivent administration, or in subjects 30 minutes after ingestion of a GLT-containing formulation. (E) Plasma cortisol concentrations in control subjects or in rhinitis patients who received one week of Narivent treatment (1 puff/nostril twice a day). *P<0.05. **P<0.01. ***P<0.001; analysis of variance plus Tukey post hoc test.

• 

  Fig. 2 High mobility group protein box 1 (HMGB1) release and effect of glycyrrhizin on survival of different cultured leukocytes. (A) HMGB1 levels in the media of different human leukocyte types purified from peripheral blood and cultured for 12 hours. (B) Effect of GLT on cell survival of different human leukocyte types purified from peripheral blood and cultured for 24 hours. Columns represent the mean±standard error of the mean of at least three experiments conducted in duplicate. **P<0.01. ***P<0.001; analysis of variance plus Tukey post hoc test. PI, propidium Iodide.

Reference

1. Pallier C, Scaffidi P, Chopineau-Proust S, Agresti A, Nordmann P, Bianchi ME, et al. Association of chromatin proteins high mobility group box (HMGB) 1 and HMGB2 with mitotic chromosomes. Mol Biol Cell. 2003; 8. 14(8):3414–3426. PMID: 12925773.

2. Ulloa L, Messmer D. High-mobility group box 1 (HMGB1) protein: friend and foe. Cytokine Growth Factor Rev. 2006; 6. 17(3):189–201. PMID: 16513409.

3. Yang H, Wang H, Czura CJ, Tracey KJ. HMGB1 as a cytokine and therapeutic target. J Endotoxin Res. 2002; 8(6):469–472. PMID: 12697092.

4. Rouhiainen A, Tumova S, Valmu L, Kalkkinen N, Rauvala H. Pivotal advance: analysis of proinflammatory activity of highly purified eukaryotic recombinant HMGB1 (amphoterin). J Leukoc Biol. 2007; 1. 81(1):49–58. PMID: 16980512.

5. Cavone L, Muzzi M, Mencucci R, Sparatore B, Pedrazzi M, Moroni F, et al. 18β-glycyrrhetic acid inhibits immune activation triggered by HMGB1, a pro-inflammatory protein found in the tear fluid during conjunctivitis and blepharitis. Ocul Immunol Inflamm. 2011; 6. 19(3):180–185. PMID: 21426233.

6. Dumitriu IE, Baruah P, Manfredi AA, Bianchi ME, Rovere-Querini P. HMGB1: guiding immunity from within. Trends Immunol. 2005; 7. 26(7):381–387. PMID: 15978523.

7. Bianchi ME, Manfredi AA. Immunology: dangers in and out. Science. 2009; 3. 323(5922):1683–1684. PMID: 19325105.

8. Lotze MT, Tracey KJ. High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat Rev Immunol. 2005; 4. 5(4):331–342. PMID: 15803152.

9. Agnello D, Wang H, Yang H, Tracey KJ, Ghezzi P. HMGB-1, a DNA-binding protein with cytokine activity, induces brain TNF and IL-6 production, and mediates anorexia and taste aversion. Cytokine. 2002; 5. 18(4):231–236. PMID: 12126646.

10. Mollica L, De Marchis F, Spitaleri A, Dallacosta C, Pennacchini D, Zamai M, et al. Glycyrrhizin binds to high-mobility group box 1 protein and inhibits its cytokine activities. Chem Biol. 2007; 4. 14(4):431–441. PMID: 17462578.

11. Fulkerson PC, Rothenberg ME. Targeting eosinophils in allergy, inflammation and beyond. Nat Rev Drug Discov. 2013; 2. 12(2):117–129. PMID: 23334207.

12. Salpietro C, Cuppari C, Grasso L, Tosca MA, Miraglia Del Giudice M, La Rosa M, et al. Nasal high-mobility group box-1 protein in children with allergic rhinitis. Int Arch Allergy Immunol. 2013; 161(2):116–121. PMID: 23343652.

13. Pittelli M, Felici R, Pitozzi V, Giovannelli L, Bigagli E, Cialdai F, et al. Pharmacological effects of exogenous NAD on mitochondrial bioenergetics, DNA repair, and apoptosis. Mol Pharmacol. 2011; 12. 80(6):1136–1146. PMID: 21917911.

14. Lapucci A, Pittelli M, Rapizzi E, Felici R, Moroni F, Chiarugi A. Poly (ADP-ribose) polymerase-1 is a nuclear epigenetic regulator of mitochondrial DNA repair and transcription. Mol Pharmacol. 2011; 6. 79(6):932–940. PMID: 21441600.

15. Damiani V, Camaioni A, Viti C, Scire AS, Morpurgo G, Gregori D. A single-centre, before-after study of the short- and long-term efficacy of Narivent(®) in the treatment of nasal congestion. J Int Med Res. 2012; 40(5):1931–1941. PMID: 23206477.

16. Olukoga A, Donaldson D. Liquorice and its health implications. J R Soc Promot Health. 2000; 6. 120(2):83–89. PMID: 10944880.

17. Rosenberg HF, Dyer KD, Foster PS. Eosinophils: changing perspectives in health and disease. Nat Rev Immunol. 2013; 1. 13(1):9–22. PMID: 23154224.

18. Lotfi R, Herzog GI, DeMarco RA, Beer-Stolz D, Lee JJ, Rubartelli A, et al. Eosinophils oxidize damage-associated molecular pattern molecules derived from stressed cells. J Immunol. 2009; 10. 183(8):5023–5031. PMID: 19794066.