摘要
目的探讨培养神经元缺氧后蛋白激酶A(PKA)活性变化及其与神经元缺氧凋亡的关系。方法建立体外培养W istar大鼠胎鼠皮层神经元模型及培养神经元缺氧模型,用不同浓度的PKA抑制剂(Rp-cAMP),在不同的缺氧时相点观察神经元细胞膜和细胞质中PKA的活性、Bc l-2和TUNEL表达的规律。结果随着缺氧时间的延长,培养神经元细胞膜PKA的活性显著增加,Bc l-2表达降低,TUNEL荧光染色阳性率及平均荧光强度显著升高。而应用Rp-cAMP后细胞凋亡情况显著减轻。结论PKA和Bc l-2均参与了缺氧神经元凋亡;PKA抑制剂Rp-cAMP可减轻缺氧神经元凋亡,且该作用与Bc l-2表达降低有关;PKA的激活在缺氧神经元凋亡中起促发作用。
Objective To explore the relationship between changes of activity of protein kinase A （PKA） and cell apoptosis in cultured neuron after cell hypoxia. Methods Based on the model of rat cortical neuron in vitro, we added four different concentration of RpcAMP, which specifically inhibit PKA, into the culture media of four different hypoxic time group every 2 hours. Then we observed the expression of Bcl-2 and the situation of neuron apoptosis. Results With the prolonging of hypoxic time, the membrane PKA activity obviously increased and simultaneously Bcl-2 expression decreased, the positive rate of fluorescence staining and the average fluorescent intensity of TUNEL significantly increased. Pre - conditioning of Rp-cAMP reversed the Bcl-2 expression reduce and neuron apoptosis increase. Conclusion PKA and Bcl-2 are involved in the process of neuronal apoptosis after hypoxia. Rp-cAMP can restrain the hypoxic neuronal apoptosis through the expression of Bcl-2. Activation of PkA can be harmful to cultured neuron during hypoxia procedure.
出处
《局解手术学杂志》
2006年第3期166-168,共3页
Journal of Regional Anatomy and Operative Surgery
基金
国家自然科学基金资助项目(39670269)
