神经酰胺诱导小鼠皮层神经元凋亡

Ceramide Induces Apoptosis in Culture Cortical Neurons of Fetal Mice

本实验以原代培养的小鼠大脑皮层神经元为模型，观察了天然神经酰胺在神经元凋亡中的作用。从测定ＬＤＨ漏出率、ＭＴＴ代谢率、ＤＮＡ凝胶电泳，Ｇｉｅｍｓａ染色以及电镜观察等各个方面，探讨了神经酰胺对神经元的作用模式。研究发现，神经酰胺在５００～１０００ｎＭ浓度范围内，作用１２ｈ以上，即可诱导原代培养的皮层神经元凋亡，而且此作用具有时间依赖性和剂量依赖性。表明神经酰胺不仅对ＨＬ－６０细胞有促凋亡的作用，对大脑皮层神经元同样具有促凋亡的作用。

In this study, the effect of natural ceramide has been tested on primary cultures of cortical neurons. In order to study if the cortical neurons undergoing apoptosis, some markers such as the leakage rate of lactate dehydrogenase(LDH) and the MTT metabolic rate were assessed, also the morphological changes were observed by Giemsa staining. DNA ladders were determined by gel electrophoresis, as well as the cell ultrastructurales were observed under electronmicroscope. The results showed that 500 to 1000nM of ceramide could induce apoptosis of culture cortical neurons in dose-dependent and time-dependent way after 12 hrs of incubation. The ultrastructural changes are consistent with the changes observed by Giemsa staining or DNA ladders on gel electrophoresis.