The study aimed to examine the roles of anxiety and intolerance of uncertainty (IU) in conditioned fear learning under an uncertain context induced by the contingency reversal of the association between the conditioned stimulus and the unconditioned stimulus (CS-US). The study sample comprised 53 participants, randomly divided into two groups: a non-instruction group and an instruction group. The experimental procedure encompassed five stages: pre-acquisition, acquisition, generalization, reversal acquisition, and reversal generalization. Our study primarily focused on analyzing a moderated mediation model. In the instructed group, we observed that the reversed fear generalization response was directly influenced by the pre-reversal fear generalization response, while also being indirectly mediated by the IU factor. However, in the non-instructed group, we did not find a significant mediating effect of IU. Moreover, we noted that the mediation of IU was contingent on the instructional information. It is noteworthy that anxiety did not exhibit a discernible role in conditioned fear within the uncertainty condition in our study. The findings provide novel insights into fear-related phenomena, emphasizing the intricate interplay between individual traits and fear generalization under conditions of uncertainty. They contribute to understanding the mechanisms of emotional and cognitive interactions in uncertain conditions.