The fatigue tests were conducted for cold-drawn eutectoid steels having different activation energies and microstructures to investigate the mechanism of fatigue strength degradation by irreversible hydrogen. The fatigue strength of a sample with low activation energy was decreased by irreversible hydrogen, whereas the fatigue strength of a sample with high activation energy was not. When the activation energy for irreversible hydrogen becomes small, the desorption of irreversible hydrogen from its trap site is easily induced by cyclic loading, which results in a decrease in fatigue strength. When a sample having high activation energy was annealed at 473 K, the fatigue strength was decreased by irreversible hydrogen although the activation energy barely changed. This is because the precipitation of fine carbides due to annealing causes higher sensitivity to hydrogen embrittlement. High sensitivity to hydrogen embrittlement was concluded to induce the fatigue strength degradation by irreversible hydrogen. Therefore, the degradation of fatigue strength by irreversible hydrogen depends on the activation energy and sensitivity to hydrogen embrittlement.