Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
권호 표지

Anti-Inflammatory Effects of Poly-${\gamma}$-Glutamic Acid on DNCB-Induced Allergic Contact Dermatitis in Dogs

개에서 DNCB에 의해 유발된 알레르기성 접촉피부염에 대한 폴리감마글루탐산의 항염증 효과

  • Kim, Hyun-Gon (College of Veterinary Medicine, Kyungpook National University) ;
  • Kim, Kil-Soo (College of Veterinary Medicine, Kyungpook National University) ;
  • Oh, Tae-Ho (College of Veterinary Medicine, Kyungpook National University)
  • Accepted : 2012.08.20
  • Published : 2012.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

Allergic contact dermatitis (ACD) is an inflammatory skin disease and regarded as a prototype of T-cell mediated delayed-type hypersensitivity reactions. Poly-${\gamma}$-glutamic acid (PGA) is a biodegradable polymer that is produced by Bacillus subtilis. This study was performed to assess the effects of PGA in a canine model of ACD. ACD was induced on the back of dogs induced by sensitization and repeated application by 2,4-dinitro-1-chlorobenzene (DNCB). Topical treatment of PGA was applied once a day for 12 days and skin biophysical parameters including transepidermal water loss (TEWL), skin hydration, skin pH, skin thickness and erythema index, were measured every two days during experimental periods. Histopathology and immunohistochemistry were performed to evaluate the antiinflammatory effect. In skin biophysical parameters, TEWL, skin hydration, skin thickness and erythema index were significantly increased, with a maximum increase appeared on day 2 (p < 0.05). On the other hand, skin pH was significantly decreased, with a maximum decrease appeared on day 2 (p < 0.01). After the completion of PGA treatment, skin biophysical parameters were significantly reached those of baseline in a time-dependent manner (p < 0.05). In histopathology, marked increases of epidermal thicknesses were induced after DNCB challenge with numerous inflammatory cell infiltrations and edematous changes, decreases of connective tissue occupied regions in dermis. In addition, marked increases of cytokine - tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$)-immunoreactivities in the dermis and of apoptotic markers - caspase-3 and PARP-immunoreactivities in the epidermis were observed in DNCB-PBS control as compared with intact control, respectively (p < 0.01). It means, the ACD and related apoptotic changes were induced by DNCB in the present study. However, these ACD induced by DNCB and related apoptosis in epidermis were significantly inhibited by treatment of PGA treated skin, the decreases of infiltrated inflammatory cells and related decreases of pro-inflammatory cytokine immunoreactivities were also observed (p < 0.01). Based on these findings, PGA may have anti-inflammatory and alleviatory effects in the allergic contact dermatitis.

알레르기성 접촉피부염은 T세포와 대식세포가 관여하는 세포매개성 면역반응으로 항원에 노출된 뒤 수 일 후에 증상이 나타나는 지연형 반응이다. 그 과정은 감작기와 유발기로 나뉘는데 감작기에는, 표피장벽을 통해 유입된 항원이 표피기저층에 있는 항원전달세포에 의해 처리된 후 림프절로 이동되어 T세포에 의해 인식되고 그 T세포는 항원특이 T세포로 활성화된다. 유발기는 동일 항원이 재감작될 때 항원특이 T세포의 반응을 활성화시키고 다양한 cytokine 분비를 통해 염증세포를 항원 유입부위로 이동시킨다. 본 연구에서는 DNCB로 알레르기성 접촉피부염을 유발한 개의 모델에 폴리감마글루탐산의 항염증 효과를 평가하였다. 폴리감마글루탐산은 12일간 적용하였고 실험기간 동안 이틀 간격으로 피부 생리학적 지표를 측정하였으며 적용 후 cytokine 측정과 조직병리학적 검사를 실시하였다. DNCB 적용후 피부 생리학적 지표의 변화로 표피경유수분손실, 피부 수화도, 피부 두께 그리고 홍반지수는 증가하였고 피부 산도는 감소하였다(p < 0.05). 조직병리학적 검사결과 염증세포 침윤과 부종성 변화에 의한 상피두께 증가 및 진피 결합조직의 감소가 특징적으로 나타났다. 또한 진피에서 pro-inflammatory cytokine인 TNF-${\alpha}$와 IFN-${\gamma}$ 수치 및 상피에서 apoptotic change의 지표인 caspase-3와 PARP 면역반응세포의 수치가 유의적으로 증가하였다(p < 0.01). 하지만 폴리감마글루탐산 적용으로 피부 생리학적 지표(p < 0.05) 및 조직병리학적 변화가(p < 0.01) 기본 수치로 회복되었다. 따라서 본 연구를 통해 개에서 DNCB에 의한 알레르기성 접촉피부염 유발 및 폴리감마글루탐산의 우수한 항염증 효과를 확인하였고 그 결과 폴리감마글루탐산은 향후 피부염에 대한 치료제로 사용할 수 있을 것으로 기대된다.

Keywords

References

  1. Assuma R, Oates T, Cochran D, Amar S, Graves DT. IL-1 and TNF antagonists inhibit the inflammatory response and bone loss in experimental periodontitis. J Immunol 1998; 160: 403-409.
  2. Berardesca E, Distante F, Vignoli GP, Oresajo C, Green B. Alpha hydroxyacids modulate stratum corneum barrier function. Br J Dermatol 1997; 137: 934-938. https://doi.org/10.1111/j.1365-2133.1997.tb01554.x
  3. Beutler B, Greenwald D, Hulmes JD, Chang M, Pan YC, Mathison J, Ulevitch R, Cerami A. Identity of tumor necrosis factor and the macrophage-secreted factor cachectin. Nature 1985; 316: 552-554. https://doi.org/10.1038/316552a0
  4. Biziuleviciene G, Puidokaite G, Siaurys A, Mauricas M. An anti-inflammatory effect of murine fetal liver cells in BALB/c mouse contact hypersensitivity model. Int Immunopharmacol 2007; 7: 744-749. https://doi.org/10.1016/j.intimp.2007.01.012
  5. Chai OH, Lee HK, Lee YC, Lee MS, Han EH, Kim HT and Song CH. Roles of $TNF-\alpha$ and IgE in the late phase of contact hypersensitivity induced by trimellitic anhydride. Exp Mol Med 2005; 37: 408-417. https://doi.org/10.1038/emm.2005.51
  6. Chen WY. Function of hyaluronan in wound repair. Wound Repair Regen 2002; 7: 79-89.
  7. Chris Pickard, Andrew M. Smith, Hywel Cooper, Ian Strickland, John Jackson, Eugene Healy and Peter S. Friedmann. Investigation of Mechanisms Underlying the T-Cell Response to the Hapten 2,4-Dinitrochlorobenzene. J Invest Dermatol 2007; 127: 630-637. https://doi.org/10.1038/sj.jid.5700581
  8. Cole KK, Perez-Polo JR. Poly(ADP-ribose) polymerase inhibition prevents both apoptotic-like delayed neuronal death and necrosis after H2O2 injury. J Neurochem 2002; 82: 19-29. https://doi.org/10.1046/j.1471-4159.2002.00935.x
  9. De Jager MW, Gooris GS, Dolbnya IP, Bras W, Ponec M. Novel lipid mixtures based on synthetic ceramides reproduce the unique stratum corneum lipid organization. J Lipid Res 2004; 45: 923-932. https://doi.org/10.1194/jlr.M300484-JLR200
  10. De Vry CG, Valdez M, Lazarov M, Muhr E, Buelow R, Fong T, Iyer S. Topical application of a novel immunomodulatory peptide, RDP58, reduces skin inflammation in the phorbol ester-induced dermatitis model. J Invest Dermatol 2005; 125: 473-481. https://doi.org/10.1111/j.0022-202X.2005.23831.x
  11. Dunstan RW, Herdt TH, Olivier B, Mei BS, Credille KM, Kennis RA, Maier RL, Oliver B, Castle S, Reinhart GA, Davenport GM. Age- and breed-related differences in canine skin surface lipids and pH. Advanced in veterinary dermatology 2002; 4: 37-42.
  12. Fluhr JW, Kao J, Jain M, Ahn SK, Feingold KR, Elias PM. Generation of free fatty acids from phospholipids regulates stratum corneum acidification and integrity. J Invest Dermatol 2001; 117: 44-51. https://doi.org/10.1046/j.0022-202x.2001.01399.x
  13. Friedmann PS. Contact sensitisation and allergic contact dermatitis: immunobiological mechanisms. Toxicol Lett 2006; 162: 49-54. https://doi.org/10.1016/j.toxlet.2005.10.008
  14. Hachem JP, Crumrine D, Fluhr J, Brown BE, Feingold KR, Elias PM. pH directly regulates epidermal permeability barrier homeostasis, and stratum corneum integrity/cohesion. J Invest Dermatol 2003; 121: 345-353. https://doi.org/10.1046/j.1523-1747.2003.12365.x
  15. Hayashi T, Hara S, Hasegawa K. Enhanced contact hypersensitivity by delayed T-helper 2 response in BALB/c mice. Allergy Asthma Proc 2009; 30: 449-457. https://doi.org/10.2500/aap.2009.30.3240
  16. Hwang SJ, Ku SK, Lee KW, Oh TH, Oh WS. Effect of Topical Application of Intercellular Lipids on Sodium Lauryl Sulphate-Damaged Skin Barrier Function in Dogs. J Vet Clin 2008; 25: 330-339.
  17. Hecker KH, Niizeki H, Streilein JW. Distinct roles for transforming growth factor-beta2 and tumour necrosis factoralpha in immune deviation elicited by hapten-derivatized antigen-presenting cells. Immunology 1999; 96: 372-380. https://doi.org/10.1046/j.1365-2567.1999.00684.x
  18. Hideki Watanabe, Olivier Gaide, Virginie Petrilli, Fabio Martinon, Emmanuel Contassot, Stephanie Roques, Jean A. Kummer, Jurg Tschopp and Lars E. French. Activation of the IL-1b-Processing Inflammasome Is Involved in Contact Hypersensitivity. J Invest Dermatol 2007; 127: 1956-1963. https://doi.org/10.1038/sj.jid.5700819
  19. JL Ríos, E Bas and MC Recio. Effects of Natural Products on ContactDermatitis. Curr Med Chem 2005; 4: 65-80.
  20. JW Fluhr, R Darlenski, I Angelova-Fischer, N Tsankov, D Basketter. Skin Irritation and Sensitization: Mechanisms and New Approaches for Risk Assessment. Skin Pharmacol Physiol 2008; 21: 191-202. https://doi.org/10.1159/000135635
  21. Kao JS, Fluhr JW, Man MQ, Fowler AJ, Hachem JP, Crumrine D, Ahn SK, Brown BE, Elias PM, Feingold KR. Short-term glucocorticoid treatment compromises both permeability barrier homeostasis and stratum corneum integrity: inhibition of epidermal lipid synthesis accounts for functional abnormalities. J Invest Dermatol 2003; 120: 456-464. https://doi.org/10.1046/j.1523-1747.2003.12053.x
  22. Kermer P, Liman J, Weishaupt JH, Bahr M. Neuronal apoptosis in neurodegenerative diseases: from basic research to clinical application. Neurodegener Dis 2004; 1: 9-19. https://doi.org/10.1159/000076665
  23. Kimber I, Holliday MR, Dearman RJ. Cytokines and the regulation of allergic sensitization to chemicals. Toxicol Lett 1995; 82: 491-496.
  24. Kim DY, Jung JA, Kim TH, Seo SW, Jung SK, Park CS. Oral administration of Uncariae rhynchophylla inhibits the development of DNCB-induced atopic dermatitis-like skin lesions via IFN-gamma down-regulation in NC/Nga mice. J Ethnopharmacol 2009; 122: 567-572. https://doi.org/10.1016/j.jep.2008.12.029
  25. Kim KS, Ahn MJ, Kim GS, Park SC, Rhee MH, In JG, Kim GK, Nah YL, Kim HH, Han SH. Effects of Opuntia humifusa Extract on DNCB-induced Allergic Contact Dermatitis in BALB/c Mice. Lab Anim 2007; 23: 169-173.
  26. Kim TH, Jung JA, Kim GD, Jang AH, Ahn HJ, Park YS, Park CS. Melatonin inhibits the development of 2,4-dinitrofluorobenzene- induced atopic dermatitis-like skin lesions in NC/Nga mice. J Pineal Res 2009; 47: 324-329. https://doi.org/10.1111/j.1600-079X.2009.00718.x
  27. Kolbe L, Kligman AM, Schreiner V, Stoudemayer T. Corticosteroid-induced atrophy and barrier impairment measured by non-invasive methods in human skin. Skin Res Technol 2001; 7: 73-77. https://doi.org/10.1034/j.1600-0846.2001.70203.x
  28. Kupper TS. Immune and inflammatory processes in cutaneous tissues. J Clin Invest 1990; 86: 1783-1789. https://doi.org/10.1172/JCI114907
  29. Lecart S, Boulay V, Raison-Peyron N, Bousquet J, Meunier L, Yssel H, Pene J. Phenotypic characterization of human CD4+ regulatory T cells obtained from cutaneous dinitrochlorobenzene-induced delayed type hypersensitivity reactions. J Invest Dermatol 2001; 117: 318-325. https://doi.org/10.1046/j.1523-1747.2001.01403.x
  30. Leung DYM, Bleber T. Atopic Dermatitis. The Lancet 2003; 361: 151-160. https://doi.org/10.1016/S0140-6736(03)12193-9
  31. Manome H, Aiba S, Tagami H. Simple chemicals can induce maturation and apoptosis of dendritic cells. Immunology 1999; 98: 481-490. https://doi.org/10.1046/j.1365-2567.1999.00916.x
  32. Marlene Bonneville, Cyril Chavagnac, Marc Vocanson, Aurore Rozieres, Josette Benetiere, Ingrid Pernet, Alain Denis, Jean- Francois Nicolas and Ana Hennino. Skin Contact Irritation Conditions the Development and Severity of Allergic Contact Dermatitis. J Invest Dermatol 2007; 127: 1430-1435. https://doi.org/10.1038/sj.jid.5700726
  33. Nagai H, Matsuo A, Hiyama H, Inagaki N, Kawada K. Immunoglobulin E production in mice by means of contact sensitization with a simple chemical, hapten. J Allergy Clin Immunol 1997; 100: 39-44. https://doi.org/10.1016/S0091-6749(97)70192-1
  34. Narins RS, Bowman PH. Injectable skin fillers. Clin Plast Surg 2005; 32: 151-162. https://doi.org/10.1016/j.cps.2004.12.002
  35. Nixon R, Frowen K, Moyle M. Occupational dermatoses. Aust Fam Physician 2005; 34: 327-333.
  36. Niwa S, Ochi T, Hirano Y, Wang T, Inagaki N, Shudo K, Nagai H. Effect of Am-80, a retinoid derivative, on 2,4-dinitrofluorobenzene-induced contact dermatitis in mice. Pharmacology 2000; 60: 208-214. https://doi.org/10.1159/000028371
  37. Olmos A, Giner RM, Recio MC, Rios JL, Cerda-Nicolas JM, Manez S. Effects of plant alkylphenols on cytokine production, tyrosine nitration and inflammatory damage in the efferent phase of contact hypersensitivity. Br J Pharmacol 2007; 152: 366-373.
  38. Ross R, Reske-Kunz AB, The role of NO in contact hypersensitivity. Int Immunopharmacol 2001; 1: 1469-1478. https://doi.org/10.1016/S1567-5769(01)00091-1
  39. Saary J, Qureshi R, Palda V, Dekoven J, Pratt M, Skotnicki-Grant S, Holness L. A systematic review of contact dermatitis treatment and prevention. J Am Acad Dermatol 2005; 53: 845-855.
  40. Smulson ME, Pang D, Jung M, Dimtchev A, Chasovskikh S, Spoondle A, Simbulan-Rosenthal C, Rosenthal D, Yakovlev A, Dritschilo A. Irreversible binding of poly(ADP)ribose polymerase cleavage product to DNA ends revealed by atomic force microscopy: possible role in apoptosis. Cancer Res 1998; 58: 3495-3498.
  41. Sung MH, Park C, Kim CJ, Poo H, Soda KJ, Makoto Ashiuchi. Natural and Edible Biopolymer Poly-g-glutamic Acid: Synthesis, Production, and Applications. Chem Rec 2005; 5: 352-366. https://doi.org/10.1002/tcr.20061
  42. Tewari M, Quan LT, O'Rourke K, Desnoyers S, Zeng Z, Beidler DR, Poirier GG, Salvesen GS, Dixit VM. Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmAinhibitable protease that cleaves the death substrate poly (ADP-ribose) polymerase. Cell 1995; 81: 801-809. https://doi.org/10.1016/0092-8674(95)90541-3
  43. Trucco C, Oliver FJ, de Murcia G, Menissier-de Murcia J. DNA repair defect in poly(ADP-ribose) polymerase-deficient cell lines. Nucleic Acids Res 1998; 26: 2644-2649. https://doi.org/10.1093/nar/26.11.2644
  44. Ueda Y, Sone T, Inagaki N, Nagai H. Effects of prednisolone on the cutaneous reaction and skin barrier function in mice treated with a hapten. Biol Pharm Bull 2003; 26: 618-621. https://doi.org/10.1248/bpb.26.618