The present disclosure discloses an anti-fatigue mat and methods for making the same. The method for making an anti-fatigue mat includes providing a leather comprising a first surface and a second surface opposite to the first surface, forming a color layer on the first surface, cutting the leather, putting the cut leather into a mold, injecting an elastomeric material into the mold, curing the elastomeric material to form an elastic layer on the second surface and a plurality of concave structures corresponding to the plurality of protruding structures on the first surface, and polishing the first surface to remove the color layer without caving towards the second surface. The anti-fatigue mat can have anti-fatigue performance and vivid stereoscopic impression, where being simple to fabricate with low costs and can be industrialized.