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目的 鸡胚内注射甲氨喋呤（ＭＴＸ）， 制作鸡胚神经管缺陷动物模型。利用该模型， 进一步研究观察病变脊髓的病理变化及组织化学改变。方法 选用上海农科院“莱杭”鸡受精蛋６６ 枚，在孵化期第４、５ 天， 注射ＭＴＸ， 计量为：０．０１ ｍｇ／ｋｇ 及０ ．０２ ｍｇ／ｋｇ。孵化３ 周后， 选取病变鸡， 采用ＮＡＤＰＨ黄递酶组织化学染色， 测定病变脊髓神经元ＮＯＳ含量； 电镜观察病变脊髓神经元内线粒体结构变化。结果 ＮＡＤＰＨ黄递酶组织化学染色显示神经管缺陷病变脊髓出现ＮＯＳ阳性染色神经元， 神经元数量减少， 形态发育差。电镜观察， 病变脊髓线粒体肿胀， 嵴减少。结论 神经管缺陷病变脊髓有ＮＯ产生，ＮＯ对神经细胞有毒性作用， 使线粒体功能受损， 能量代谢障碍。
Objective To investigate the effect of methotrexate (MTX) injected into chicken embryos on the animal model of neural tube defects in chick embryo. Using this model, we further studied the pathological changes and histochemical changes of the diseased spinal cord. Methods Sixty 66 eggs of “Lai Hang” chicken from Shanghai Academy of Agricultural Sciences were selected and injected MTX at the fourth and fifth day of hatching period. The dosage was 0.01 mg / kg and 0. 02 mg / kg. After incubation for 3 weeks, the diseased chickens were selected and NADPH-diaphorase histochemical staining was performed to determine the content of NOS in the diseased spinal cord neurons. The mitochondrial structure of spinal cord neurons was observed under electron microscope. Results NADPH diaphorase histochemical staining showed that NOS positive stained neurons were found in the spinal cord of neural tube defects, the number of neurons decreased and the morphology was poorly differentiated. Electron microscopy, lesions of the mitochondria swelling of the spinal cord, crest reduction. Conclusion There is NO production in the spinal cord of neural tube defects and NO is toxic to nerve cells, impaired mitochondrial function and impaired energy metabolism.