In a mouse model of skin repair we found that the class I-IIa histone deacetylase inhibitor (DI) Trichostatin A (TSA) accelerated tissue regeneration. Unexpectedly, this effect was suppressed by Sirtinol, a class III HDACs (sirtuins) selective inhibitor. The role of sirtuins (SIRTs) was then investigated by using Resveratrol and a novel SIRT1-2-3 activator, the MC2562 compound we recently synthesized. Both Resveratrol and MC2562 were effective in accelerating wound repair. The local administration of natural or synthetic SIRT activators, in fact, significantly accelerated skin regeneration by increasing keratinocyte proliferation. In vitro experiments revealed that the activation of SIRTs stimulated keratinocyte proliferation via eNOS phosphorylation and NO production. In this condition, the class I member HDAC2 was found S-nitrosylated on cysteine, a posttransduction modification associated to loss of activity and DNA binding capacity. After DIs or SIRT activators treatment chromatin immunoprecipitation (ChIPs) showed, in fact, a significant HDAC2 detachment from the promoter region of Insulin Growth Factor I (IGF-I), Fibroblast Growth Factor 10 (FGF-10) and Epithelial Growth Factor (EGF) which may be the final recipients and effectors of the SIRT-NO-HDAC signaling cascade. Consistently, the effect of SIRT activators was reduced in the presence of L-NAME, a general inhibitor of NO synthesis. In conclusion, the NO-dependent crosstalk among class III and I histone deacetylases suggests an unprecedented signaling pathway important for skin repair.