Effect of Ketogenic Diet on Pentylenetetrazole (PTZ) Kindled Rats with Medial Prefrontal Cortex Damage: Behavioral, Morphological and In Silico Analysis

Abstract

Ketogenic diet (KD) has gained attention as an adjunct therapy for refractory epilepsy due to its anticonvulsant and neuroprotective effects. This study investigated the neuroprotective potential of different KD formulations in a pentylenetetrazol kindling (PTZK (35 mg/kg) every other day for 20 days. To promote kindling, PTZ was increased by 5 mg/kg in subsequent intraperitoneal administrations after the fourth injection. Rats were fed various diets 10 days before Kindling and during the Kindling. Thirty (30) young adult female Wistar rats were divided into six groups (n=5 per group). Group 1, Control (normal feed); Group 2, PTZK (35 mg/kg); Group 3, PTZK + levetiracetam (10 mg/kg); Group 4, PTZK + palm-kernel oil-KD; Group 5, PTZK + castor oil-KD; and Group 6, PTZK + olive-KD. Seizure severity was assessed using Racine’s scoring, cognitive function was evaluated using the novel object-recognition test. Number of neurons in the supralimbic medial prefrontal cortex (smPFC) layer 2/3 was estimated using unbiased stereological methods, including histology and physical dissector technique. DNA-methylation was assessed using Feulgen reaction and molecular docking analysis evaluated the binding-affinities of ketone-bodies and levetiracetam. Results demonstrate that Ketone-bodies' strong bond to HCR2 and NMDA receptor may protect against neurodegeneration as KD-treated groups showed reduced seizure severity, delayed seizure onset, and improved behavioral outcomes compared to the PTZK group. Number of neurons and DNA integrity were preserved in KD groups, almost similar to control group, compared to marked neurodegeneration in smPFC layer 2/3 of PTZK group. KDs provide neuroprotection against PTZK-neurodegeneration in smPFC layer 2/3.