Background: The differentiation of human epidermal keratinocyte is one of the essential factors for skin barrier. Adiponectin is well established for its beneficial aspects such as anti-inflammation, anti-senescence and regulation of SIRT1. In human epidermal keratinocytes, SIRT1 reports to induce differentiation by its upregulation. However, the relationship of adiponectin and keratinocyte differentiation is still unrevealed. Objectives: This study aimed to analyze the effect of adiponectin on the induction of differentiation in human epidermal keratinocytes via regulation of calcium influx. Methods: Normal human epidermal keratinocytes (NHEKs) knocked down for adiponectin receptor and SIRT1 gene by siRNA were respectively treated with recombinant human adiponectin, nifedipine and Ca2+. Calcium influx was measured by Fura-2/AM dye. Levels of NHEKs differentiation markers were examined via immunoblot analysis. Results: The silence of SIRT1 decreased calcium influx induced by adiponectin. Stimulated keratinocyte differentiation markers by adiponectin were suppressed by SIRT1 siRNA while calcium channel blocker showed no significant effect. Conclusion: In this study, the results suggest a role for adiponectin as an induction agent of keratinocyte differentiation to improve and maintain keratinocyte function that forms the skin barrier.