Ensete glaucum seeds, being known as one of traditional medicine, are commonly used in folk medicine to treat urinary stones, edema, and osteoarthritis-related problems. However, no scientific evidence has been reported to support these uses. This study focused on investigating the antinociceptive and antiurolithiatic activities of E. glaucum seed aqueous extract (EGE). The antinociceptive effect of EGE was evaluated in mice using thermal (hot plate test) and chemical (acetic acid and formalin-induced nociception test) pain models at various doses (50, 100, 200, 400 mg/kg; p.o.). The anti-urolithiatic activity of the EGE (200, 400 mg/kg; p.o.) was assessed in the sodium glyoxylate-induced urolithiasis in mice and in vitro nucleation and aggregation assays. The analgesic effect of EGE was detected at several doses via peripheral and central antinociceptive mechanisms in the acetic acid-induced writhing (50-400 mg/kg), hot plate (100-200 mg/kg), and formalin-induced licking (200-400 mg/kg) tests. EGE had potential against urolithiasis through its ability to modify several serum and urine biochemical parameters on glyoxylate-induced nephrolithiasis. The extract at the dose of 400 mg/kg significantly improved the inflammatory cells, kidney tissue structure, and renal calcification. The extract also exhibited significant anti-urolithiatic, anti-inflammatory, and antioxidant activities in some in vitro models. These outcomes suggest that E. glaucum aqueous seed extract possesses antinociceptive activity and may aid in the prevention of urinary stones. Further studies are needed to elucidate the effectiveness of E. glaucum seeds in the analgesic activity and management of urolithiasis disease.