NUCB2/nesfatin-1 has been implicated in various physiological functions such as feeding, drinking and the sleep-wake cycle, but its effect on epileptic activity is not clear. The aim of this study was to investigate the effects of NUCB2/nesfatin-1 on penicillin-induced epileptic activity and to answer the question of whether only NUCB2/nesfatin-1 cause epileptic activity in rats. Thirty-five rats were randomly assigned to the following seven groups: control, saline, penicillin, two different doses of NUCB2/nesfatin-1 (100 and 300 pmol), penicillin and two different doses of NUCB2/nesfatin-1 (100 and 300 pmol). EEGs were recorded before the injections and during the following 120 min. The EEG and motor findings of epileptic seizures were observed in NUCB2/nesfatin-1-injected groups. NUCB2/nesfatin-1 significantly increased the EEG power spectrum, spike frequency and amplitude values in penicillin-induced epileptic rats. These findings indicate that NUCB2/nesfatin-1 both causes epileptic activity and increases penicillin-induced epileptic activity in rats.