Background: This study aimed to evaluate the role of beta-blocker therapy on modulating interleukin (IL)-33/ST2 (interleukin-1 receptor-like 1) signaling during ventricular remodeling related to heart failure (HF) after acute myocardial infarction (AMI). Methods: Sprague-Dawley rats that survived surgery to induce AMI were randomly divided into the placebo group and the beta-blocker treatment group. A sham group was used as a control. Left ventricular (LV) function variables, the myocardial infarct size, fibrosis and IL-33/ST2 protein expression was measured. Results: Compared with the placebo group, b-blocker treatment significantly improved LV function and reduced infarct size (p < 0.05). There was higher protein expression of IL-33 (p < 0.05) and sST2 (p < 0.05), as well as higher expression of fibrosis (p < 0.05), compared to the sham group. Notably, the high expression of cardioprotective IL-33 was not affected by b-blocker treatment (p > 0.05), however, treatment with b-blocker enhanced IL-33/ST2 signaling, with lower expression of sST2 (p < 0.05) and significantly attenuated fibrosis (p < 0.05). Conclusions: Our study suggested that b-blocker therapy might play a beneficial role in the modulation of IL-33/ST2 signaling during ventricular remodeling. These results may be helpful in identifying IL-33/ST2 systems as putative b-blocker targets at an early stage after AMI.