Primary afterdischarge in organotypic hippocampal slice cultures: Effects of standard antiepileptic drugs
Primary afterdischarge in organotypic hippocampal slice cultures: Effects of standard antiepileptic drugs
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DOI：
10.1111/j.1528-1167.2012.03597.x
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发表时间：
2012-11-01
期刊：
影响因子：
5.6
通讯作者：
Heinemann, Uwe
中科院分区：
文献类型：
--
作者：
Albus, Klaus;Wahab, Abdul;Heinemann, Uwe
目的：在海马体中，强烈的高频电刺激引起持久的节律同步（初级后放电）。为了检查器官型海马切片培养物（OHSC）中原发性后放电（PAD）作为诱发癫痫发作的体外模型的适用性，我们详细计算了 PAD 对标准抗癫痫药物（AED）的敏感性，并将必要的浓度与对部分和全身强直阵挛性癫痫动物模型有效的浓度进行了比较。方法：根据 采用界面培养法对 8 至 11 日龄的 Wistar 大鼠进行实验。 CA1 中的 PAD 是通过以引发最大群体峰值所需强度两倍的强度刺激辐射层来引发的。确定了卡马西平、苯妥英、丙戊酸、苯巴比妥、安定和乙舒酰亚胺对 PAD 的持续时间和频率特性以及 PAD 的强直样和阵挛样细分的影响，并根据浓度响应曲线计算了 AED 效力的半最大有效浓度 (EC50) 值。 苯妥英、丙戊酸、苯巴比妥和地西泮可缩短 PAD 的持续时间以及 PAD 的强直样和阵挛样细分。效果是浓度依赖性的并且是可逆的。乙昔酰亚胺无效。不同 AED 对 PAD 细分的影响不同。卡马西平和苯妥英以相似的比例缩短强直样和阵挛样细分，而苯巴比妥、安定和丙戊酸优先缩短阵挛样细分。低浓度的地西泮可延长类似强直细分的持续时间，这是其他 AED 所未见的效果。 AED 对强直样和阵挛样细分的频率特性的抑制作用是可变的，并且仅在较高浓度下观察到。意义：卡马西平和苯妥英在 OHSC 的 PAD 测试中比在最大电击和点燃癫痫发作测试中更有效。苯巴比妥、安定和丙戊酸在 PAD 测试中的有效性与它们在 MES 测试中的有效性相匹配，但丙戊酸和安定引发癫痫发作测试除外。丙戊酸在 OHSC 中的效果不如点燃癫痫发作试验中的效果，而地西泮在该动物模型中比在 OHSC 中更有效（全身性癫痫发作）或更少（局灶性癫痫发作和放电后持续时间）。我们的结论是，OHSC 中的 PAD 测试是诱发癫痫发作的合适体外模型。该模型可以作为初步筛选，以确定最有希望的先导化合物，以便在体内功效和毒性模型中进一步评估和表征。
Purpose: In the hippocampus intense high frequency electrical stimulation induces a long-lasting rhythmic synchronization (primary afterdischarge). In order to examine the suitability of primary afterdischarges (PADs) in organotypic hippocampal slice cultures (OHSCs) as an in vitro model of evoked seizures, we have worked out in detail the sensitivity of PADs to standard antiepileptic drugs (AEDs) and compared the necessary concentrations to those that were effective in animal models of partial and generalized tonicclonic seizures.Methods: OHSCs were prepared according to the interface culture method from 8 to 11-day-old Wistar rats. A PAD in CA1 was elicited by stimulating the stratum radiatum with an intensity of two times that required to elicit a maximal population spike. The effects of carbamazepine, phenytoin, valproic acid, phenobarbital, diazepam, and ethosuximide on the duration and on frequency properties of PADs and the tonic-like and clonic-like subdivisions of PADs were determined, and as a measure of the AEDs potency half maximal effective concentration (EC50) values were calculated from concentrationresponse curves.Key Finding: Carbamazepine, phenytoin, valproic acid, phenobarbital, and diazepam reduced the durations of PADs and tonic-like and clonic-like subdivisions of PADs. The effects were concentration dependent and reversible. Ethosuximide was ineffective. The effects on subdivisions of PADs differed between AEDs. Carbamazepine and phenytoin shortened the tonic-like and clonic-like subdivisions at similar proportions, whereas phenobarbital, diazepam, and valproic acid preferentially shortened the clonic-like subdivision. Diazepam at low concentrations increased the duration of tonic-like subdivisions, an effect not seen with the other AEDs. The suppressive effects of AEDs on frequency properties of tonic-like and clonic-like subdivisions were variable and observed only at higher concentrations.Significance: Carbamazepine and phenytoin were more effective in the PAD test in OHSCs than in the maximal electroshock and kindled seizures tests. The effectiveness of phenobarbital, diazepam, and valproic acid in the PAD test matched their effectiveness in the MES test andwith the exception of valproic acid and diazepamin kindled seizures tests. Valproic acid was less effective in OHSCs than in the kindled seizure tests, and diazepam was more (generalized seizures) or less (focal seizures and afterdischarge durations) effective in this animal model than in OHSCs. We conclude that the PAD test in OHSCs is a suitable in vitro model of evoked seizures. The model could serve as an initial screen to identify the most promising leads for further evaluation and characterization in in vivo models of efficacy and toxicity.