A novel dietary therapy for epilepsy: potential mechanisms of action

项目总结/摘要 超过三分之一的癫痫患者即使在服用 多种抗癫痫药物（AEDs）。不受控制的强直-阵挛性癫痫发作使患者处于高风险中 癫痫猝死（SUDEP）是癫痫患者死亡的主要原因， 抗药性癫痫高脂肪，非常低碳水化合物生酮饮食（KDs）帮助一些 癫痫不受控制的患者，尤其是儿童，但被描述为难以接受 并且患者通常不耐受，导致高的不依从率。 我们已经从Dravet综合征（DS）的小鼠模型中获得了初步数据，这是一种毁灭性的疾病。 儿童期发作的癫痫伴难治性癫痫发作和SUDEP的高发病率，并发现 通过在他们的饮食中补充乳清（WD），可以大大减少饥饿引起的死亡。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KD增加5-HT水平和5-HT， 释放到老鼠的大脑中。这一点很重要，因为5-HT已知可以抑制癫痫发作， 在控制呼吸方面起着关键作用。5-HT系统中的缺陷与这两种情况有关 呼吸功能障碍和SUDEP。在这个项目中，我们将测试以下假设：1） 乳清通过减少癫痫发作频率和/或刺激癫痫发作后的 呼吸通过增加脑5-HT，和2）乳清防止发作后死亡，通过逆转的基本 DS小鼠的潜在缺陷，这被认为是GABA能快- 尖峰中间神经元。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 高血压诱发癫痫发作和高效液相在体微透析 通过色谱法测量杏仁核（与癫痫发作有关的区域）中细胞外的5-HT 传播和发作后呼吸暂停。我们将对海马脑进行膜片钳记录 来自WD喂养的DS小鼠的切片，以评估中间神经元的内在电生理特性 和锥体神经元，并将确定是否由于WD介导的兴奋性变化 增加5-HT信号。这些研究将提供临床前数据， 临床试验，以评估牛奶乳清补充剂作为一种安全和健康的饮食疗法， 耐药性癫痫，比KD更可口和有效。这些实验可能 也为针对牛奶参与的β-肾上腺素能机制的治疗开辟了新的途径 哇。

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.