Background: beta-carotene is an essential antioxidant, providing protection against type 2 diabetes mellitus, cardiovascular illnesses, obesity, and metabolic syndrome. This study investigates the impact of beta-carotene on biochemical parameters and pancreatic insulin expression in mice exposed to ethanol. Methods: Thirty-six C57BL/6 mice (Mus musculus) were divided into six groups: 1. C (control), 2. LA (3% alcohol dose), 3. MA (7% alcohol dose), 4. B (0.52 mg/kg body weight/day beta-carotene), 5. LA+B (3% alcohol dose + 0.52 mg/kg body weight/day beta-carotene), and 6. MA+B (7% alcohol dose plus 0.52 mg/kg body weight/day beta-carotene). After 28 days, the animals were euthanized for serum and pancreatic tissue collection. Biochemical analysis and pancreatic insulin expression were performed. One-way ANOVA was used. Results: The B, LA+B, and MA+B groups improved insulin levels and decreased HOMA-beta versus the C group, with the LA+B and MA+B groups also showing lower ADH and ALDH levels than their nonsupplemented counterparts (p < 0.05). The B, LA+B, and MA+B groups showed a greater beta-cell mass area compared to the unsupplemented groups. Additionally, the LA+B and MA+B groups demonstrated significantly increased beta-cell area and integrated optical density compared to the LA and MA groups, respectively (p < 0.001). Conclusions: In mice, beta-cell loss led to increased glucose release due to decreased insulin levels. beta-carotene appeared to mitigate ethanol's impact on these cells, resulting in reduced insulin degradation when integrated optical density was used. These findings suggest that antioxidant supplementation may be beneficial in treating ethanol-induced type 2 diabetes in animal models.