The experimental protocol aimed to assess the influence of nucleotides and/or β-glucan on the production performance, growth-related gene expression in the broiler chickens’ liver tissue, intestinal histomorphology, and some biochemical parameters. One hundred forty-four newly hatched chicks were categorized into four groups based on the supplements: a control group, a group supplemented with nucleotides (200 mg/kg diet), a group with β-glucan (1 g/kg diet), and a group with both nucleotides and β-glucan. The study's findings showed that, when compared to the control group, all dietary supplemented groups showed a substantial (P < 0.05) improvement in production parameters without having a negative impact on the general health of broilers. Additionally, the treatments positively impacted the expression of some genes associated with growth performance in the broiler chickens’ liver tissue, such as the insulin-like growth factor 1 (IGF-1) and growth hormone receptor (GHR). The supplemented groups also demonstrated a significant (P < 0.05) enhancement in intestinal histomorphology, characterized by increased villi length, crypt depth, and goblet cell number. Furthermore, dietary supplementation of nucleotides and/or β-glucan led to a significant (P < 0.05) elevation in total protein and albumin levels, with no significant effect on globulin, AST, and ALT. The conclusion can be drawn that adding nucleotides along with β-glucan to the diet led to enhancements in growth performance, the expression of genes related to growth (GHR and IGF-1), intestinal histomorphology, and certain biochemical parameters (specifically total protein and albumin).