Emerging evidence indicates a pathogenic role of protracted neuroinflammation in the various neurodegenerative diseases, including epilepsy. Neuroinflammation may contribute to neuronal hyperexcitability underlying seizure formation. The current research aims to examine the changes in the levels of proinflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the penicillin epilepsy model. In the present study, 12 male Wistar albino rats were randomly divided into two groups as sham and penicillin epilepsy model. Seizures were induced with the intracortical (i.c.) single microinjection 500 IU of penicillin-G into neocortex. Rats were decapitated after observing the cortical epileptic activity and brains were removed by craniotomy. Proinflammatory cytokines (TNF-α and IL-1β) were measured by using ELISA methods in the cortical and hippocampal brain regions. Penicillin significantly up-regulated the expression of IL‑1β and TNF-α in the rat cortex, but did not affect the hippocampal cytokines levels. This study is indicative of the neuroinflammatory potential of cortical penicillin administration.