A novel dietary therapy for epilepsy: potential mechanisms of action

项目摘要/摘要 超过三分之一的癫痫患者即使服用药物也不能有效控制癫痫发作。 多种抗癫痫药物(AED)。不受控制的强直-阵挛发作使患者处于高危状态 癫痫猝死(SUDEP)是导致癫痫患者死亡的主要原因 抗药性癫痫。高脂肪、极低碳水化合物的生酮饮食(KDS)有助于 患有失控癫痫的患者，特别是儿童，但被描述为令人不快 而且往往是患者不能容忍的，导致高违规率。 我们已经从Drave氏综合症(DS)的小鼠模型中获得了初步数据，这是一种毁灭性的 儿童发作的癫痫伴难治性发作和SUDEP的高发生率，并发现 通过在他们的饮食中补充牛奶乳清(WD)，癫痫导致的死亡大大减少。这 WD与两种不同的KD一样具有保护作用，但不需要增加酮体。 此外，我们的初步研究表明，WD和KDS可增加5-羟色胺水平和5-羟色胺 在小鼠脑内释放。这一点很重要，因为众所周知，5-羟色胺可以抑制癫痫发作，也可以发挥作用 在控制呼吸方面起着关键作用。5-羟色胺系统中的缺陷与这两种疾病有关 呼吸功能障碍和SUDEP。在这个提议的项目中，我们将检验以下假设：1) 乳清通过减少癫痫发作频率和/或刺激癫痫发作后死亡来预防癫痫导致的死亡 通过增加大脑5-羟色胺的呼吸，以及2)乳清通过逆转基本的 DS小鼠的潜在缺陷，被认为是GABA能快速兴奋性的下降- 刺激性的中间神经元。为了确定WD如何影响癫痫发作、呼吸和神经化学， 我们将使用视频-EEG/EKG来评估WD对自发性和自发性癫痫的抗惊厥作用 高温致痫和高效液体体内微透析 用层析法测定与癫痫发作有关的杏仁核细胞外5-羟色胺 传播和后遗症呼吸暂停。我们将在海马脑上进行膜片钳记录 WD喂养DS小鼠脑片评价中间神经元的内在电生理特性 和锥体神经元，并将确定WD引起的兴奋性变化是否参与 通过增加5-羟色胺信号。这些研究将提供临床前数据，可能指导未来 评价补充乳清作为一种安全健康的饮食疗法的临床试验 比KD更可口、更有效的抗药性癫痫。这些实验可能 也导致了针对牛奶参与的5-羟色胺能机制的新的治疗途径 乳清。

Project Summary/Abstract More than one-third of people with epilepsy have inadequate control of seizures even when taking multiple antiepileptic drugs (AEDs). Uncontrolled tonic-clonic seizures place patients at high risk of sudden unexpected death in epilepsy (SUDEP), a major cause of mortality in patients with drug-resistant epilepsy. The high-fat, very low-carbohydrate ketogenic diets (KDs) help some patients with uncontrolled epilepsy, especially children, but have been described as unpalatable and are often not tolerated by patients, leading to high rates of non-compliance. We have obtained preliminary data from a mouse model of Dravet Syndrome (DS), a devastating childhood-onset epilepsy with refractory seizures and a high incidence of SUDEP, and found seizure-induced death is greatly reduced by supplementing their diet with milk whey (WD). This WD was just as protective as two different KDs, but did not require an increase in ketone bodies. Moreover, our preliminary studies suggest that a WD and KDs increase 5-HT levels and 5-HT release in the brain of mice. This is important, as 5-HT is known to inhibit seizures and also plays a critical role in control of breathing. Defects in the 5-HT system have been linked to both respiratory dysfunction and SUDEP. In this proposed project, we will test the hypotheses that 1) whey prevents seizure-induced death by reducing seizure frequency and/or stimulating postictal breathing via increased brain 5-HT, and 2) whey prevents postictal death by reversing the basic underlying defect in DS mice, which is thought to be a decrease in excitability of GABAergic fast- spiking interneurons. To determine how a WD affects seizures, breathing and neurochemistry, we will use video-EEG/EKG to evaluate the anticonvulsant effects of a WD on spontaneous and hyperthermia-induced seizures, and in vivo microdialysis with high performance liquid chromatography to measure extracellular 5-HT in the amygdala, a region implicated in seizure propagation and postictal apnea. We will perform patch clamp recordings on hippocampal brain slices from WD-fed DS mice to evaluate intrinsic electrophysiological properties of interneurons and pyramidal neurons, and will determine if changes in excitability due to the WD are mediated by increased 5-HT signaling. These studies will provide preclinical data that may guide future clinical trials to evaluate milk whey supplementation as a safe and healthy dietary therapy for drug-resistant epilepsy that is more palatable and effective than the KD. These experiments may also lead to new avenues of treatment targeted at the serotonergic mechanisms engaged by milk whey.