摘要
目的研究体外培养的神经细胞缺血缺氧性损伤后凋亡相关蛋白Bcl-2和Bax的表达,及应用MAPKs信号通路阻断剂U0126对其表达的调节作用。方法取出生1d的SD大鼠,断头处死后迅速解剖分离其皮层组织进行培养,3d后进行缺血缺氧处理致细胞损伤。并分为加药组和缺氧组,前者给予U0126,后者给予空白的DMSO溶剂作为对照。采用MTT法、免疫细胞化学、免疫蛋白印迹(western blot)技术,检测在缺氧性损伤急性期神经细胞的活力、凋亡相关蛋白Bcl-2和Bax的表达情况,及给予U0126后对神经细胞的活力、凋亡相关蛋白Bcl-2和Bax表达的影响。结果缺氧损伤后Bcl-2和Bax表达升高,细胞活力不佳,死亡较多。给予U0126后ERK1/2和ELK1磷酸化水平降低,同时Bax的表达水平降低,Bcl-2表达增高,细胞的状态和活力亦明显改善。结论给予MAPKs阻断剂U0126可以调节缺氧损伤后凋亡相关蛋白Bcl-2和Bax的表达．抑制细胞死亡,改善细胞状态。
Objective To study the effect of MAPKs signal pathway blocker U0126 on the expressions of apoptosis-related proteins, Bcl-2 and Bax, in neurons after hypoxic injury in vitro. Methods Brains from 1 day postnatal SD rats were taken for in vitro neuron culture. Three days later, the cultures were divided randomly into 3 groups： drug-treated, hypoxia and normal groups. Those in drug-treated and hypoxia groups were subjected to hypoxia and then the former was treated with 10 μmol/L U0126 dissolved with DMSO for 30 min while the latter accepted DMSO as control. Two days later, immunocytochemistry and Westem-blot were used to detect the expression of Bcl-2/Bax, as well as the phosphorylation of ERK1/2 and Elkl, both of which are key proteins of MAPKs signal transduction pathway. Results After hypoxic injury, Bcl-2/Bax expression rose and neurons displayed a poor viability with an increasing number of died cells. Compared with hypoxia group, Bcl-2 expression in drug-treated group showed an obvious increase, while Bax expression in drug-treated group decreased （P〈0.01）. Meanwhile, phosphorylation of ERK1/2 and Elk-1 in drug-treated group was lower than that in ischemia group. Conclusion U0126 can regulate the expressions of apoptosis-associated proteins, Bcl-2 and Bax, and improve the cell viability after neuronal hypoxic injury.
出处
《中华神经医学杂志》
CAS
CSCD
2006年第9期883-886,共4页
Chinese Journal of Neuromedicine
基金
上海市重点基金(024119037)上海市引进海外高层次留学人员专项基金(20040105)
关键词
缺氧损伤
凋亡
丝裂原活化蛋白激酶
Hypoxic injury
Apoptosis
Mitogen activated protein kinases
