Restorative Dentistry and Endodontics

The Korean Academy of Conservative Dentistry (대한치과보존학회)
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The effect of neuropeptides on secretion of Interleukin-8(IL-8)

Interleukin-8 (IL-8) 분비에 미치는 neuropeptides의 영향에 관한 연구

  • Kim, Kyung-Jun (Department of Conservative Dentistry, Division of Dentistry, Graduate School, Kyunghee University) ;
  • Park, Sang-Hyuk (Department of Conservative Dentistry, Division of Dentistry, Graduate School, Kyunghee University) ;
  • Choi, Kyoung-Kyu (Department of Conservative Dentistry, Division of Dentistry, Graduate School, Kyunghee University) ;
  • Park, Sang-Jin (Department of Conservative Dentistry, Division of Dentistry, Graduate School, Kyunghee University)
  • 김경준 (경희대학교 대학원 치의학과 치과보존학교실) ;
  • 박상혁 (경희대학교 대학원 치의학과 치과보존학교실) ;
  • 최경규 (경희대학교 대학원 치의학과 치과보존학교실) ;
  • 박상진 (경희대학교 대학원 치의학과 치과보존학교실)
  • Published : 2006.05.01
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Abstract

We investigated the secretion of Interleukin-8 (IL-8) from ginviva and periodontal ligament stimulated with Substance P (SP) and Calcitonin Gene-related Peptide (CGRP). Gingiva (GF), periodontal ligament (PDLF) and pu)p (PF) tissues were collected from extracted intact 3rd molars. Cultured cells were stimulated with different concentrations of SP for 4 hrs, and stimulated with SP, CGRP and Tumor Necrosis Factor-$\alpha$ (TNF-$\alpha$) for 8 hrs. Then RNase Protection Assay was carried out. ELISA was performed using supernatants of stimulated cells for quantitative analysis of IL-8. Results were assessed using student t-test with significance of P<0.05. According to this study, the results were as follows: 1. IL-8 mRNA was detected in all type of cells studied (PF, GF and PDLF) 2. IL-8 mRNA expression was not increased after stimulating 4 hrs with SP ($10^{-5}M$) and SP ($10^{-8}M$) compared with Mock stimulation in all type of cells studied. 3. IL-8 mRNA expression was not increased after stimulating 8 hrs with SP ($10^{-4}M$) and CGRP ($10^{-6}M$) compared with Mock stimulation in all type or cells studied. 4. TNF-$\alpha$ (2 ng/ml) increased the expression of IL-8 mRNA in all kind of cells studied. 5. The secretion of IL-8 from GF was increased 8 hrs after the stimulation with CGRP ($10^{-6}M$)(p<0.05). 6. The secretion of IL-8 from PDLF was. increased 8 hrs after the stimulation with SP ($10^{-4}M$)(p<0.05). Calcitonin Gene-related Peptide (CGRP) increased Interleukin-8 (IL-8) which plays an important role in chemotaxis of neutrophil in Calcitonin Gene-related Peptide (CGRP) gingival tissue , whereas Substance P increased the secretion of IL-8 from periodontal ligament.

본 연구는 치수조직, 치은, 치주인대로부터 배양된 조직을 SP (Substance P)로 4시간 SP, CGRP (Calcitonin Gene-related Peptide) , Tumor necrosis factor-$\alpha$ (TNF-$\alpha$)로 8시간 자극 후 RNase Protection Assay를 시행하고, IL-8의 분비량을 측정해 다음 결과를 얻었다. 1. IL-8 mRNA는 모든세포에서 발현됐다. 2. IL-8 mRNA 발현은 SP ($10^{-5}M$)와 SP ($10^{-8}M$)로 4시간 자극 시 증가되지 않았다. 3. IL-8 mRNA 발현은 SP ($10^{-4}M$)와 CGRP ($10^{-6}M$)로 8시간 자극 시 증가되지 않았다. 4. TNF-$\alpha$ (2 ng/ml) 자극 시, IL-8 mRNA 발현이 증가됐다. 5. 치은 세포를 CGRP ($10^{-6}M$)로 8시간 자극 시, IL-8 분비량이 증가했다 (p<0.05). 6. 치주인대 세포를 SP ($10^{-4}M$)로 8시간 자극 시 IL-8 분비량이 증가했다 (p<0.05).

Keywords

References

  1. Kiss M. Kemeny L. Gyulai R, Michel G. Husz S. Kovacs R. Dobozy A. Ruzicka T. Effects of the neuropeptides substance P, calcitonin gene-related peptide and a-melanocyte-stimulating hormone on the IL-8/ IL-8 receptor system in a cultured human keratinocyte cell line and dermal fibroblasts. Inflammation 23:557-567, 1999 https://doi.org/10.1023/A:1020294507767
  2. Bartold PM. Kylstra A. Lawson R. Substance P. An Immunohistochemical and Biochemical Study in Human Gingival Tissues. A Role for Neurogenic Inflammation? J Periodonto 165:1113-1121, 1994
  3. Rameshwar P, Gascon P. Induction of negative hematopoietic regulators by neurokinin-A in bone marrow stroma. Blood 88:98-106, 1996
  4. Baggiolini M, Walz A, Kunkel SL. Neutrophil-activating peptide-1 /Interleukin 8. a novel cytokine that activates neutrophils. J Clin Invest 84: 1045-1049, 1989 https://doi.org/10.1172/JCI114265
  5. Kent LW, Dyken RA, Rahemtulla F, Allison AC, Michalek SM. Effects of in vitro passage of healthy human gingival fibroblasts on cellular morphology and cytokine expression. Archs oral Bio 141:263-270. 1996
  6. Guo CJ. Lai JP, Luo HM. Douglas SD. Ho WZ. Substance P up-regulates macrophage inflammatory protein-1$\beta$ expression in human T lymphocytes. J Neuroimmunol 131: 160-167, 2002 https://doi.org/10.1016/S0165-5728(02)00277-1
  7. Huang GTJ, Potente AP, Kim JW. Chugal N. Zhang X. Increased interleukin-8 espression in inflamed human dental pulps. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 88:214-220, 1999 https://doi.org/10.1016/S1079-2104(99)70118-6
  8. Fitzgerald JE, Kreutzer DL. Localization of Interleukin-8 in human gingival tissues, Oral Microbiol and Immunol 10:297-303, 1996
  9. Larsen CG. Anderson AO. Appella E, Oppenheim JJ, Matsushima K. The neutrophil-activating protein (NAP-I) is also chemotactic for T lymphocytes. Science 243:1464-1466, 1989 https://doi.org/10.1126/science.2648569
  10. Peveri P. Walz A. Dewald B. Baggiolini M. A novel neutrophil activating factor produced by human mononuclear phagocytes. J Exp Med 167:1547-1559, 1988 https://doi.org/10.1084/jem.167.5.1547
  11. Cuesta MC. Quintero L. Pons H. Suarez-Roca H. Substance P and calcitonin gene-related peptide increase IL-1/$\beta$, IL-6 and $TNF^{a}$ secretion from human peripheral blood mononuclear cells, Neurocheistry Int 40:301-306, 2002 https://doi.org/10.1016/S0197-0186(01)00094-8
  12. Park SH, Hsiao GYW, Huang GTJ. Role of substance P and calcitonin gene-related peptide in the regulation of interleukin-8 and monocyte chemotactic protein-l expression inhuman dental pulp. Int Endod J 37: 185-192, 2004 https://doi.org/10.1111/j.0143-2885.2004.00782.x
  13. Bordin S, Narayanan S, Reddy J, Cleveland D. Page RC. Fibroblast subtypes in the periodontium. J Periodont Res 19:642-644, 1984 https://doi.org/10.1111/j.1600-0765.1984.tb01332.x
  14. McCulloch CAG. Bordin S. Role of fibroblast subpopulations in periodontal physiology and pathology. J Periodont Res 26: 144-154, 1991 https://doi.org/10.1111/j.1600-0765.1991.tb01638.x
  15. Heyeraas KJ. Berggreen E. Interstitial fluid pressure in normal and inflamed pulp. Crit Rev Oral Biol Med 10(3): 328-336, 1999 https://doi.org/10.1177/10454411990100030501
  16. Berggreen E. Heyeraas KJ. Effect of the sensory neuropeptide antagonists h-CGRP( (8-37)) and SR 140.33 on pulpal and gingival blood flow in ferrets. Arch Oral Bio 145(7):537-542, 2000
  17. Fistad I. K vinnsland IH, Jonsson R. Heyeraas KJ. Effect of intermittent long-lasting electrical tooth stimulation on pulpal blood flow and immunocompetent cells: a hemodynamic and immunohistochemical study in young rat molars. Exp Neurol 146(1) :230-239, 1997 https://doi.org/10.1006/exnr.1997.6523
  18. Fristad I. Heyeraas KJ, Kwinnsland IH. Jonsson R. Recruitment of immunocompetent cells after dentinal injuries in innervated and denervated young rat molars: an innunohistochemcal study. J Histochem Cytochem 43(9) :871-879, 1995 https://doi.org/10.1177/43.9.7543913
  19. Bongenhielm D, Haegerstrand A. Theodorsson E. Fried K. Effect of neuropeptides on growth of cultivated rat molar pulp fibroblasts. Regul Pept 60(2-3): 91-8, 1995 https://doi.org/10.1016/0167-0115(95)00115-8
  20. Patel T. Park SH, Lin LM, Chiappelli F, Huang GTJ. Substance P induces interleukin-8 secretion from human dental pulp cells. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 96:478-485, 2003 https://doi.org/10.1016/S1079-2104(03)00036-2
  21. Ansel JC. Armstrong CA. Song I. Quinlan KL. Olerud JE, Caughman SW, Bunnett NW. Interaction of the skin and nervous system. J Investig Dermatolo Symp Proc 2(1) :23-26, 1997
  22. Raap T. Justen HP, Miller LE. Cutolo M. Scholmerich J. Straub RH. Neurotransmitter modulation of interleukin 6(1L-6) and IL-8 secretion of synovial fibroblasts in patients with rheumatoid arthritis compared to osteoarthritis. J Rheumatol 27: 2558-2565, 2000
  23. Detmers PA, Lo SK, Olsen-Egbert E. Walz A. Baggiolini M. Cohn ZA. Neutrophil-activating protein l/interleukin 8 stimulates the binding activity of the leukocyte adhesion receptor CD11b /CD18 on human neutrophils. J Exp Med 117: 1155-1162, 1990
  24. Rot A. Neutrophil attractant/ activation protein-1(interleukin-8) induces in vitro neutrophil migration by haptotactic mechanism. Eur J Immunol 23:303-306, 1993 https://doi.org/10.1002/eji.1830230150
  25. Takashiba S, Takigawa M, Takahashi K. Myokai F, Nishimura F. Chihara T, Kurihara H. Nomura Y. Murayama Y. Interleukin-8 is a major neutrophil chemotactic factor derived from cultured human gingival fibroblasts stimulated with interleukin-1/$\beta$ or tumor necrosis factor alpha. Infection and Immunity 60: 5253-5258, 1992