The study investigated the potential antiurolithiatic effects of an aqueous extract from dried Punica granatum leaves. The extract, obtained using the Soxhlet extraction method, yielded less than 22%. Acute toxicity studies established the LD 50 values and safety of the extract, indicating no harmful effects even at concentrations up to 2000 mg/kg body weight. A starting therapeutic dose of 1/10 th of the LD 50 dose (200 mg) was chosen, setting a dosage range of 100 to 400 mg/kg body weight for subsequent experiments. Male rats were selected to induce urolithiasis due to the similarity of their urinary system to humans. The study induced urolithiasis in rats by administering ethylene glycol, effectively replicating urinary stone formation observed in humans. Microscopic examination highlighted the extract’s potential to impede crystal growth, a critical factor in urolithiasis. Urine volume studies emphasized potential kidney function disruption in the calculi-induced group, while groups treated with Cystone and the test phytochemical showed increased urine output, potentially aiding in flushing out calculi. The study further explored body weight changes, urinary pH, routine urine analysis, crystal formation, calcium and oxalate excretion, and serum biochemical analysis. Treatment with the aqueous extract at specific doses prevented an increase in critical serum markers associated with kidney damage, suggesting a potential protective effect against nephritic damage. Histopathological examination revealed the aqueous extract’s potential to mitigate calcium oxalate depositions and associated renal tissue abnormalities. Additionally, the extract exhibited nephroprotective potential against gentamicin-induced nephrotoxicity, underlining its broader protective effects on renal health.