Vitamin E (E) was previously reported to reduce CaOx crystal formation. This study explored if its deficiency affects intra-renal oxidative stress and accelerates crystal deposition in hyperoxaluria. The control group (C) of rats received a standard diet and drinking water, while the experimental groups received 0.75% ethylene glycol (EG) in drinking water for 42 days. Of the latter, one group received a standard diet (EG group), one received a low E (LE) diet (EG+LE group), and the last received an LE diet with vitamin E supplement (4 mg) (EG+LE+E group). The C+LE and C+LE+E groups were the specific controls for the last two experimental groups, respectively. In a separate experiment, EG and EG+LE rats were studied on days 3-42 to examine the temporal relationship between oxidative change and crystal formation. Urinary biochemistry and activity/levels of anti-oxidative and oxidative enzymes in the glomeruli and tubulo-interstitial specimens (TIS) were examined. In EG rats, CaOx crystal accumulation was associated with low anti-oxidative enzyme activity in TIS with increased oxidative enzyme expression in the glomeruli. In the EG+LE group, marked changes in anti-oxidative and oxidative enzyme levels were seen and correlated with massive CaOx deposition and tubular damage. The increased oxidative stress seen with EG+LE treatment was largely reversed by vitamin E supplementation. Temporal study showed that decrease in antioxidative defense and free radical formation in the EG+LE group occurred before crystal deposition. This study shows that LE disrupts the redox balance and causes cell death, thereby favoring crystal formation.