Hot work tools steels, which are submitted to severe solicitations in service, are often damaged by thermal fatigue and corrosion. Interrupted Thermal Fatigue experiments were performed on X38CrMoV5 tool steel between 100 and 650 °C, in air and reduced oxygen partial pressure atmospheres after primary or secondary vacuum. A thermal and thermo-mechanical analysis by finite element method was carried out to estimate the strains and stresses undergone by the axisymmetric disc-shaped specimen during thermal cycling. The morphology, structure and phase composition of the oxide layer were analysed using scanning electron microscopy, energy dispersive spectroscopy and X-ray diffraction. It was shown that the thickness, homogeneity, structure, and compacity of the oxide multilayer formed on the specimen changed depending on test atmosphere. In air, crack initiation and propagation were strongly assisted by oxidation, leading to reduced fatigue life compared to inert atmospheres. Steel softening was highlighted in sub-surface, whatever the test atmosphere.