Establishment of a ketogenic diet model and behavioral evaluation in epileptic mice
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1. Children’s Hospital of Nanjing Medical University, Nanjing 210008, China; 2. School of Public Health,Southeast University, Nanjing 210096, China; 3. School of Basic Medical Sciences, Nanjing Medical University, Nanjing 211166, China

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    Abstract:

    Objective To establish a mouse model of temporal lobe epilepsy and to investigate the effects of the ketogenic diet (KD). Specifically, we aimed to compare the differences in body mass, fur condition, and blood ketone levels between mice on the KD and those on a regular diet; and to further explore the impact of KD on seizure susceptibility, as well as depression- and anxiety-like behaviors. Methods Thirty-one 4-weeks-old male SPF-grade ICR mice ( 20 ~ 22 g) underwent acclimatization for 1 week. Eleven mice were assigned randomly to control conventional diet (CON + ND, n= 5) and KD (CON + KD, n= 6) groups. The remaining 20 mice were subjected to pilocarpine-induced status epilepticus (SE). The SE model was validated using Racine scale scoring (≥ stage 4),and successfully modeled mice were allocated randomly to epileptic standard diet (SE + ND) and epileptic ketogenic diet group (SE + KD) groups. Seizure frequency was recorded via video monitoring, and anxiety- and depression-like behaviors were assessed using open field, elevated plus maze, and forced swimming tests ( FST). Results ( 1) Establishment of epilepsy models: of the 20 mice subjected to modeling, four failed to reach Racine stage 4 and two died. The remaining 14 successfully modeled mice were allocated randomly to SE + KD (n= 7) and SE + ND (n=7) groups. During intervention, two mice died in the SE + KD group, while no mortality occurred in the SE + ND group. (2)Establishment of the ketogenic diet model: epileptic mice had lower baseline body mass than controls, but there was no significant difference in weight change between the dietary interventions ( P>0. 05). Blood ketone levels in the CON + KD group were consistently ≥ 1. 0 mmol / L at days 1, 7, 14, and 28. By day 2 of ketogenic feeding, nine mice developed fur soiling with fecal matter and diarrhea, progressing to rectal prolapse in five severe cases by day 3. CON + ND and SE + ND mice retained glossy coats and formed stools throughout. (3) Behavioral tests: SE + KD mice had fewer seizures compared with SE + ND mice. In the FST, CON + KD exhibited shorter immobility times than CON + ND mice. Conclusions Standardized KD intervention effectively maintained therapeutic ketosis (blood ketones ≥ 1. 0 mmol / L) without significant weight impact. Notably, a KD suppressed epileptic discharges and ameliorated anxiety- and depression-like behaviors.

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  • Received:March 26,2025
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  • Online: January 08,2026
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