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Background: Cyperus rotundus L. is used in various dosage forms by Unani physicians in the treatment of urolithiasis. Aims and Objectives: The present study aims to evaluate the effect of hydroalcoholic extract of Cyperus rotundus in nephrolithiatic male Sprague Dawley rats. Materials and Methods: The animals were divided into 6 groups of 6 each. Group I received regular rat food and drinking water ad libitum. Group II to VI were treated with Ethylene glycol (0.75%, V/V) and Ammonium chloride (1%, W/V) in drinking water for 7 days to induce urolithiasis. Group II was sacrificed after 7 days administration of lithogenic agents however, from 8th day, group IV was treated with Cystone (750 mg/kg) and group V and VI with hydroalcoholic extract of Cyperus rotundus (100 mg/kg and 170 mg/kg, respectively) for further 14 days. Group III left untreated after 7 days administration of lithogenic agent till 14 days and sacrificed on 22nd day. Urine, biochemical parameters, kidney homogenate analysis and histopathology were carried out. Crystalluria analysed by light microscopy. Results: The test drug at both the doses showed significant reduction (P < 0.001) in number of urinary crystals. Test groups showed significant reduction in urine sodium (P < 0.05) and calcium (P < 0.001) while increased in urine magnesium. Serum creatinine (P < 0.01) and urea (P < 0.05) level significantly reduced in test groups. Histopathology of kidney showed almost normal kidney architecture. Kidney homogenate analysis showed significant reduction (P < 0.05) of calcium in group VI. Conclusion: The results showed that test extract has significant antilithiatic effect in terms of solute balance, reduction in crystal numbers and improvement in renal cell derangement.