Calphostin C对培养神经元缺氧后Bcl-2表达及神经元凋亡的研究

目的探讨蛋白激酶C(PKC)对神经元缺氧凋亡的影响及作用机制.方法建立体外培养孕Wister大鼠皮层神经元模型及培养神经元缺氧模型,在此基础上观察神经元存活率、Bcl-2和凋亡DNA表达的规律;然后用3种不同浓度的PKC催化亚基特异性抑制剂CalphostinC预孵育培养神经元后进行缺血处理,观察神经元上述指标的改变.结果随着缺氧时间的延长和CalphostinC浓度的增加,培养神经元的存活数显著下降、Bcl-2免疫组化阳性细胞数明显减少、TUNEL荧光染色平均荧光强度显著升高.结论(1)PKC和Bcl-2均参与了缺氧神经元凋亡;(2)PKC抑制剂CalphostinC可加重缺氧神经元凋亡;(3)PKC的激活在缺氧神经元的凋亡中起保护作用,且该作用可能是通过促进Bcl-2表达实现的.

Influence of protein kinase C on the expression of Bcl-2 and anoxic cell apoptosis of cultured rat cortical neuron

Objective: To investigate the mechanism that how protein kinase C(PKC) affects the anoxic neuron apoptosis. Methods: First, in vitro models of neuronal culture and anoxic neuronal culture were established. Second, three different concentrations of Calphostin C were used to cocultured with the neurons. Finaly, the number of survival neurons and the expression of Bcl - 2 and apoptotic DNA fragments were detected in each group. Results: With hypoxic time prologation and the concentration of Calphostin C increasing, the number of survival neurons decreased obviously, the number of neurons that positively stained by Bcl - 2 decreased. And on the contrary the neurons' average optical density of TUNEL was significantly increased. Conclusions: (1) Both PKC and Bcl - 2 are participate in the process of anoxic neuron apoptosis. (2) Calphostin C can aggravate anoxic neuron apoptosis. (3) The activation of PKC can protect anoxic neuron apoptosis through promoting the expression of Bcl - 2.