Ketogenic Diet Alleviates Hippocampal Neurodegeneration Possibly via ASIC1a and the Mitochondria-Mediated Apoptotic Pathway in a Rat Model of Temporal Lobe Epilepsy

Abstract

Qi Qiao,1 Zhenzhen Qu,1 Shuang Tian,2 Huifang Cao,3 Yange Zhang,4 Can Sun,5 Lijing Jia,1,* Weiping Wang1,* 1The Department of Neurology, The Second Hospital of Hebei Medical University, Shijiazhuang, People’s Republic of China; 2The Department of Neurology, Shijiazhuang People’s Hospital, Shijiazhuang, People’s Republic of China; 3The Department of Rehabilitation, The Second Hospital of Hebei Medical University, Shijiazhuang, People’s Republic of China; 4The Department of Pediatrics, The Second Hospital of Hebei Medical University, Shijiazhuang, People’s Republic of China; 5The Department of Neurology, The Third Hospital of Peking University, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Weiping Wang; Lijing Jia, The Department of Neurology, The Second Hospital of Hebei Medical University, No. 215 Heping West Road, Shijiazhuang, 050000, People’s Republic of China, Email [email protected]; [email protected]: The ketogenic diet (KD) is a proven therapy for refractory epilepsy. Although the anti-seizure properties of this diet are understood to a certain extent, the exploration of its neuroprotective effects and underlying mechanisms is still in its infancy. Tissue acidosis is a common feature of epileptogenic foci. Interestingly, the activation of acid-sensing ion channel 1a (ASIC1a), which mediates Ca2+-dependent neuronal injury during acidosis, has been found to be inhibited by ketone bodies in vitro. This prompted us to investigate whether the neuroprotective effects induced by the KD occur via ASIC1a and interconnected downstream mechanisms in a rat model of temporal lobe epilepsy.Methods: Male Sprague-Dawley rats were fed either the KD or a normal diet for four weeks after undergoing pilocarpine-induced status epilepticus (SE). The effects of KD on epileptogenesis, cognitive impairment and hippocampal neuron injury in the epileptic rats were subsequently evaluated by video electroencephalogram, Morris water maze test and Nissl staining, respectively. The expression of ASIC1a and cleaved caspase-3 in the hippocampus were determined using Western blot analysis during the chronic period following SE. Moreover, the intracellular Ca2+ concentration, mitochondrial membrane potential (MMP), mitochondrial reactive oxygen species (mROS) and cell apoptosis of hippocampal cells were detected by flow cytometry.Results: We found that the KD treatment strongly attenuated the spontaneous recurrent seizures, ameliorated learning and memory impairments and prevented hippocampal neuronal injury and apoptosis. The KD was also shown to inhibit the upregulation of ASIC1a and the ensuing intracellular Ca2+ overload in the hippocampus of the epileptic rats. Furthermore, the seizure-induced structure disruption of neuronal mitochondria, loss of MMP and accumulation of mROS were reversed by the KD treatment, suggesting that it has protective effects on mitochondria. Finally, the activation of caspase-3 was also inhibited by the KD.Conclusion: These findings indicate that the KD suppresses mitochondria-mediated apoptosis possibly by regulating ASIC1a to exert neuroprotective effects. This may provide a mechanistic explanation of the therapeutic effects of KD.Keywords: ketogenic diet, temporal lobe epilepsy, acid-sensing ion channel 1a, mitochondrial pathway, neuroprotection

Keywords

• ketogenic diet
• temporal lobe epilepsy
• acid-sensing ion channel 1a
• mitochondrial pathway
• neuroprotection