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Recent studies regarding downstream effects of visfatin/eNAMPT in skeletal muscles have attracted much attention as the previous reports suggested this adipokine may exert insulin-mimetic effects. However, studies in vivo present conflicting data and are still controversial. In this present work, we sought to investigate whether visfatin/eNAMPT is able to reproduce insulin effects on glucose transport and lipid metabolism.We have used isolated skeletal muscles with different fibre type composition (Soleus and EDL) to examine glucose transport, GLUT-4 translocation, phosphorylation of insulin signalling pathway proteins, as well as the key parameters for fatty acid metabolism.We found that, in vitro exposure to visfatin/eNAMPT increased skeletal muscle glucose transport but only in EDL (+20%) and not in Soleus muscle (+5%). Interestingly, classical insulin signalling pathways were not significantly involved in this process. Concomitantly, visfatin/eNAMPT exerted no significant effects on muscle’s fatty acids (FA) metabolism as no change in either palmitate oxidation or esterification was observed. Importantly, combined insulin and visfatin effects were not found, suggesting non-additivity.Our data suggest that visfatin/eNAMPT plays a rather limited role in regulating skeletal muscle glucose transport and FA metabolism.