[P1-3-35] Role of the nucleotide-binding oligomerization domain-containing protein 1
pathway in the development of periodontitis
Keywords:IL-8、歯肉上皮細胞、iE-DAP
NOD1, a receptor for iE-DAP, a bacterial peptidoglycan structure, plays a role in regulating innate immunity. And it is known to induce the expression of various cytokines and antimicrobial peptides by inducing the activation of the transcription factor NF-κB, which plays an important role in immunity/inflammation, thereby acting to activate innate immunity. However, little is known about the effects of NOD1 on human squamous cell carcinoma. In this study, the effect of NOD1 on cytokine production by the gingival epithelial cell line Ca9-22 was investigated. MATERIALS AND METHODS 1) Cell culture: The gingival epithelial cell line Ca9-22 was cultured in Dulbecco's Modified Eagle's medium (high glucose) containing 10% bovine serum, 100 ug/ml penicillin, 100 μg/ml streptomycin and 2 mM L-glutamine. Ca9-22 was incubated at 5% CO2 and 37 °C. 2) After stimulation with IE-DAP (10 ng/ml) for 24 h, IL-8, IL-6, TNF-α, GMCSF and IL-1β in the supernatant were measured by ELISA. 3) Stimulation with IE-DAP (10 ng/ml) (0, 5, 10, 30, 60, 120 min) and phosphorylation of MAP kinases (p38, ERK, JNK) were confirmed by Western blotting. Results 1) Enhanced secretion of IL-8 was observed after 24 h stimulation with iE-DAP (10 ng/ml). Secretion of IL-6, TNF-α, GMCSF and IL-1β was unchanged. 2) The MAP kinases p38, ERK and JNK all showed enhanced phosphorylation upon iE-DAP (10 ng/ml) stimulation. Phosphorylation was also enhanced with time, peaking at 10 min for all three. DiscussionThese results suggest that phosphorylation of MAP kinases (p38, ERK and JNK) may play an important role in iE-DAP-induced IL-8 production in the gingival epithelium.