Formation of calcium oxalate (CaOx) crystal is associated with a higher prevalence of hypertension in a patient with impaired renal function. Vitexin, a fenugreek flavonoid, contains antioxidant, cardioprotective, and renoprotective potential. The objective was to evaluate the potential of vitexin against CaOx induced urolithiasis in uninephrectomized rats. Ethylene glycol (EG) was administered in uninephrectomized Wistar rats to induce the formation of CaOx crystals. Then, rats were administered either vehicle or vitexin (15, 30, and 60 mg/kg) for 28 days. Administration of EG-induced significant alterations in hemodynamic and electrocardiographic, urinary and serum (BUN, uric acid, creatinine, sodium, calcium, LDH, urinary citrate, albumin, and GAGs levels) in uninephrectomized rats. However, vitexin (30 and 60 mg/kg) treatment significantly attenuated these EG-induced changes. Furthermore, vitexin also significantly inhibited EG-induced elevated oxido-nitrosative stress (SOD, GSH, MDA, and NO) in renal and cardiac tissue. Up-regulated renal bikunin, iNOs, and TNF-alpha mRNA expressions whereas down-regulated OPN mRNA expression were significantly inhibited by vitexin. It also significantly reduces cardio-renal histological alterations induced by EG. In conclusion, vitexin ameliorated EG-induced urolithiasis via inhibition of GAGs, oxido-nitrosative stress, bikunin, iNOs, TNF-alpha, and upregulation of OPN expressions. Thus, vitexin can be considered as an important therapeutic moiety for the management of urolithiasis in the renal hypertensive patient.