茶碱对小脑颗粒神经元凋亡的保护作用及其作用机理

为探讨茶碱对神经元凋亡的影响及其作用机理，建立小脑颗粒神经元的低钾凋亡模型，用ＤＮＡ凝胶电泳、ＦＤＡ染色和放射免疫等方法，发现经不同浓度茶碱（１～２０ｍｍｏｌ／Ｌ）处理，低钾培养的小脑颗粒神经元存活率相应提高２２％～１００％，量效关系有高度显著性（Ｐ＜０．０１），ＥＣ５０＝２．９７ｍｍｏｌ／Ｌ；经茶碱（２０ｍｍｏｌ／Ｌ）处理的细胞，其ＤＮＡ凝胶电泳带没有“梯子”状分布；与不同浓度茶碱（５～８０ｍｍｏｌ／Ｌ）在ＣＯ２培养箱孵育１５ｍｉｎ，小脑颗粒神经元内ｃＡＭＰ均升高，且是剂量依赖性的。认为茶碱对撤去极化浓度氯化钾诱发小脑颗粒神经元的凋亡有保护作用，此作用产生除与其升高小脑颗粒神经元内ｃＡＭＰ有关外，还有其它机理。

Protective effect of theophylline and its mechanisn on apoptosis in cerebellar granule neurons

The low K + apoptotic model of cerebellar granule neurons was made to study the effect of theophylline on apoptosis in cerebellar granule neurons. Using DNA gel electrophoresis, FDA staining and radioimmuoassay methods, the survial rate of low K + cultured cerebellar granlue neurons could have been promoted from 22% to 100% as being treated with diffferent concentration theophylline (1～20 mmol/L), and the dose-effect relationship was statistically significant (P<0.01). EC 50 =2.97mmol/L. Neurons treated with 20-mmol/L-theophylline did not show 180bp ladder characteristic of endonuclear activation on its DNA gel electrophoresis. The i of the neurons rose after being incubated together with different concentration theophylline (5～80 mmol/L) for 15 minutes in CO 2 incubator and the prometed range relied on the dose of theophylline. It can be concluded that theophylline can prevent cerebellar granule neurons from apoptosis induced by depolarizing concentration of KCL deprivation in culture.