Gene expression profiles in gingival epithelium stimulated with Ergothioneine

Method: EGT was purified from Japanese Tamogitake mushroom that contains the highest amount of EDT among the mushrooms. Human Gingival Epithelium Progenitor cell (HEGP), were grown in Progenitor Cell Targeting medium (CELLNTEC). The cells were incubated with EGT at concentration of 1 mM and 24 hours. The cells incubated without EGT were used us a control. Each RNA sample extracted from the cells was reverse transcribed into cDNA. The labeled cRNA samples from cells were hybridized to whole human genome microarrays (Agilent Technologies, WD). The Ingenuity Pathway Analysis (IPA) (Ingenuity Systems, http://www.ingenuity.com) software was utilized to identify networks and pathways of interacting genes and other functional groups in the upregulated genes. Expressions of identified genes were confirmed by RT-PCR using TaqMan probes (Applied Biosystem). The results were compared between EGT stimulated cells and control. The statistical significance of the difference was analyzed using the Mann-Whitney U test. 

Result: We identified a set of 248 up-regulated genes as a result of the stimulation with EGT. Canonical pathway analysis of the up-regulated expression demonstrated that the most and 2nd most significant changes were in “Extrinsic Prothrombin Activation Pathway” and “Coagulation system”, respectively. By IPA, these were mainly via NF-kB and ERK1/2 pathway. 

Conclusion: These results indicate that EGT may affect wound healing in gingival epithelium.