J Korean Acad Conserv Dent.  2006 May;31(3):153-160. 10.5395/JKACD.2006.31.3.153.

The effect of neuropeptides on secretion of Interleukin-8 (IL-8)

• Kim KJ
• Park SH
• Choi KK
• Park SJ

• KMID: 2175814
• DOI: http://doi.org/10.5395/JKACD.2006.31.3.153

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 pulp (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 of 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.

Keyword

Interleukin-8 (IL-8); Substance P (SP); Tumor Necrosis Factor-alpha (TNF-alpha); pulp fibroblast (PF); gingival fibroblast (GF); periodontal ligament fibroblast (PDLF)

MeSH Terms

Calcitonin Gene-Related Peptide
Cells, Cultured
Chemotaxis
Enzyme-Linked Immunosorbent Assay
Gingiva
Humans
Interleukin-8*
Molar
Neuropeptides*
Neutrophils
Periodontal Ligament
Ribonucleases
RNA, Messenger
Substance P
Tumor Necrosis Factor-alpha
Calcitonin Gene-Related Peptide
Interleukin-8
Neuropeptides
RNA, Messenger
Ribonucleases
Substance P
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 RNase Protection Assay 4hrs after stimulation of the cells with MOCK and various dose of Substance P. 1. MOCK stimulation with medium only 2. Stimulation with Substance P (10-5M) 3. Stimulation with Substance P (10-8M) PF: Pulp Fibroblasts GF: Gingival Fibroblasts PDLF: Periodontal Ligament Fibroblasts

• 

  Figure 2 RNase Protection Assay 8hrs after stimulation of the cells with Mock and Substance P, CGRP, and TNF-α. 1. MOCK stimulation with medium only 2. Stimulation with Substance P (10-4M) 3. Stimulation with Calcitonin Gene Related Peptides (CGRP, 10-6M) 4. Stimulation with TNF-α (2 ng/ml) PF: Pulp Fibroblasts GF: Gingival Fibroblasts PDLF: Periodontal Ligament Fibroblasts

• 

  Figure 3 IL-8 secretion from the cells 8hrs after stimulation with SP (10-4M), CGRP (10-6M) and TNF-α (2 ng/ml). GF: Gingival Fibroblasts PDLF: Periodontal Ligament Fibroblasts

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