Umur Ahmet Sukru, Selcuki Mehmet, Bursali Adem, Umur Nurcan, Kara Burcu, Vatansever H Seda, Duransoy Yusuf Kurtulus
Faculty of Medicine, Department of Neurosurgery, Celal Bayar University, Manisa, Turkey.
Childs Nerv Syst. 2012 May;28(5):729-37. doi: 10.1007/s00381-011-1673-9. Epub 2012 Jan 14.
The aim of this study is to elucidate the preventive effect of folic acid (FA) on teratogenic effects of valporic acid (VA) in early stage chick embryos on neural tube development.
One hundred and fifty specific pathogen-free (SPF) chick eggs were used to investigate the neurulation in five groups. Group A was the control group. Group B was injected 0.02 ml of saline (0.9% NaCl) and was used for sham group. VA (0.72 mg) in 0.02 ml saline was injected in Group C, and 0.342 mcg of FA in 0.02 ml NaCl were administered to the embryos in Group D. VA (0.72 mg) + 0.342 mcg of FA in 0.02 ml saline were administered simultaneously to the eggs in Group E. At the end of 72 h, all embryos were extracted from eggs and were fixed, and for histological analyses hematoxylin and eosine was used, for detection of apoptotic cells terminal deoxyribonucleotide transferase-mediated dUTP-X nick end labeling (TUNEL) was used and for distribution of P53, bcl-2 and caspase-3, caspase-6, caspase-8 and caspase-9 immunoperoxidase techniques were used.
While there were no neural tube defects in the embryos of groups A, B and D, eight embryos died in group C and there were 12 embryos with retarded embryological development. In contrast to that, no death was observed in group E, but only eight embryos were detected with maldevelopmental delay stage.
These results suggested that VA may induce apoptotic mechanisms but not through the p53 pathway. In addition, FA effectively prevents the teratogenic influence of VA on chick embryo at neurulation stages by stopping cascade of apoptosis before caspase 3 expression.
本研究旨在阐明叶酸（FA）对丙戊酸（VA）在鸡胚早期神经管发育中致畸作用的预防效果。
使用150枚无特定病原体（SPF）鸡胚研究五组中的神经胚形成情况。A组为对照组。B组注射0.02 ml生理盐水（0.9% NaCl），用作假手术组。C组在0.02 ml生理盐水中注射0.72 mg VA，D组胚胎在0.02 ml NaCl中给予0.342 mcg FA。E组鸡蛋同时在0.02 ml生理盐水中给予VA（0.72 mg）+ 0.342 mcg FA。在72小时结束时，从鸡蛋中取出所有胚胎并进行固定，组织学分析使用苏木精和伊红染色，检测凋亡细胞使用末端脱氧核苷酸转移酶介导的dUTP-X缺口末端标记（TUNEL）法，检测P53、bcl-2以及半胱天冬酶-3、半胱天冬酶-6、半胱天冬酶-8和半胱天冬酶-9的分布使用免疫过氧化物酶技术。
A、B和D组胚胎未出现神经管缺陷，C组有8枚胚胎死亡，12枚胚胎存在胚胎发育迟缓。相比之下，E组未观察到死亡情况，但仅检测到8枚胚胎有发育延迟阶段。
这些结果表明，VA可能诱导凋亡机制，但并非通过p53途径。此外，FA通过在半胱天冬酶3表达之前阻止凋亡级联反应，有效预防了VA对鸡胚神经胚形成阶段的致畸影响。
