Due to the pleiotropic effects of ketogenic diet (KD) on pathophysiology, including inflammation and oxidative stress, medical personnels have been intensively exploring the application of it in the management of epilepsy. Meanwhile, the voltage-dependent anion channel 1 (VDAC1) is believed to be involved in regulating inflammatory and oxidative stress processes. However, the correlation between KD and VDAC1 in epilepsy has not been elucidated. Pilocarpine-induced chronic epilepsy model was utilized to examine the effects of VDAC1 antagonist VBIT-4 treatment on seizure activity, behavioral changes, inflammation, and oxidative stress under control diet (CD) and KD conditions, respectively. Chronic VBIT-4 administration mitigates seizure activity and behavioral abnormalities of epileptic mice under KD rather than CD conditions. Our further findings demonstrated that VBIT-4 treatment attenuates inflammatory response and oxidative stress in the hippocampi and serum samples of epileptic mice under KD rather than CD. Taken together, our data suggest a critical role of VDAC1 in mediating the anti-seizure efficacy of the KD, shedding light on developing novel therapeutic interventions to seizure control.