在丙戊酸致畸性大鼠模型中，宫内暴露于丙戊酸与海马细胞数量及叶酸代谢的改变有关。

Intrauterine valproate exposure is associated with alterations in hippocampal cell numbers and folate metabolism in a rat model of valproate teratogenicity.

作者信息

Semmler Alexander, Frisch Christian, Bleul Christiane, Smith Desiree, Bigler Laurent, Prost Jean-Christophe, Blom Henk, Linnebank Michael

机构信息

Swiss Epilepsy Center, Clinic Lengg and Department of Neurology, University Hospital Zurich, Zurich, Switzerland.

University Bonn, Department of Epileptology, Germany.

出版信息

Seizure. 2017 Mar;46:7-12. doi: 10.1016/j.seizure.2017.01.003. Epub 2017 Jan 27.

DOI:10.1016/j.seizure.2017.01.003

PMID:28212902

Abstract

PURPOSE

Valproate is one of the most commonly used anticonvulsive drugs. Despite its significant benefits, the teratogenicity of valproate is a relevant problem in the treatment of women of childbearing age. In addition to major congenital malformations, such as neural tube defects, reduced intelligence and attention after intrauterine valproate exposure are reported. Until now the mechanisms of teratogenicity of VPA are poorly understood and concepts how to reduce valproate teratogenicity are lacking.

METHODS

In a rat model of valproate teratogenicity we examined hippocampal cell structure in 4 week old animals with a stereological approach. As potential mechanisms of VPA teratogenicity we examined histone acetylation by western blotting and metabolites of the folate metabolism as well as global DNA methylation by tandem mass spectrometry in the brain and liver tissue of newborn pups (p0).

RESULTS

We found an increase in the number of neurons in the hippocampal areas CA1/2 (p=0.018) and CA3 (p=0.022), as well as a decreased number of astrocytes in CA1/2 (p=0.004) and CA3 (p=0.003) after intrauterine VPA exposure, as a possible indication of altered cell differentiation during intrauterine VPA exposure. Valproate exposure was also associated with an increase in 5-methyl-tetrahydrofolate (THF) (p=0.002) and a decrease in 5-10-methenyl-THF in the brain of newborn pups, as well as a reduced homocysteine plasma level (p<0.001). The described changes in hippocampal cell numbers and folate metabolism were only significant after high-dose intrauterine VPA exposure indicating a dose-dependent effect. VPA exposure was not associated with changes in histone acetylation or global DNA methylation in brain tissue in newborn pups.

CONCLUSION

This study shows that intrauterine VPA exposure is associated with changes in hippocampal cell numbers in the CA1/2 and CA3 region and in folate metabolism.

摘要

目的

丙戊酸盐是最常用的抗惊厥药物之一。尽管其益处显著，但丙戊酸盐的致畸性在育龄女性治疗中是一个相关问题。除了神经管缺陷等主要先天性畸形外，还报道了宫内暴露于丙戊酸盐后智力和注意力下降。到目前为止，丙戊酸致畸性的机制了解甚少，且缺乏降低丙戊酸盐致畸性的概念。

方法

在丙戊酸致畸性大鼠模型中，我们采用体视学方法检查了4周龄动物的海马细胞结构。作为丙戊酸致畸性的潜在机制，我们通过蛋白质印迹法检测组蛋白乙酰化，通过串联质谱法检测新生幼崽（出生0天）脑和肝组织中叶酸代谢产物以及整体DNA甲基化。

结果

我们发现，宫内暴露于丙戊酸后，海马CA1/2区（p = 0.018）和CA3区（p = 0.022）的神经元数量增加，而CA1/2区（p = 0.004）和CA3区（p = 0.003）的星形胶质细胞数量减少，这可能表明宫内暴露于丙戊酸期间细胞分化发生改变。丙戊酸暴露还与新生幼崽脑中5-甲基四氢叶酸（THF）增加（p = 0.002）、5,10-亚甲基四氢叶酸减少以及血浆同型半胱氨酸水平降低（p < 0.001）有关。所述海马细胞数量和叶酸代谢的变化仅在高剂量宫内丙戊酸暴露后显著，表明存在剂量依赖性效应。丙戊酸暴露与新生幼崽脑组织中的组蛋白乙酰化或整体DNA甲基化变化无关。