脑室周围白质软化症的体外模型中磷酸化ERK蛋白的表达及应用PD98059后对其表达的影响  

The expression of phosphorylated ERK and the effect of PD98059 in the periventricular leukomalacia model in vitro

作　　者：张晓璐[1] 张彦波[2] 李娟[1] 李新民[2] 

机构地区：[1]吉林大学公共卫生学院卫生检验教研室,吉林长春130021 [2]加拿大曼尼托巴大学医学院精神病学系神经精神药理实验室

出　　处：《中国实验诊断学》2009年第11期1497-1500,共4页Chinese Journal of Laboratory Diagnosis

摘　　要：目的观察体外构建脑室周围白纸软化症(Periventricular leukomalacia,PVL)模型的发病过程中磷酸化细胞外信号调节激酶(phosphorylated extracellular signal regulated kinase,p-ERK)的表达变化及应用ERK通路抑制剂PD98059后对其表达的影响,以探讨ERK通路是否参与PVL的发病过程。方法共分为三种干预方法:给予CG4细胞糖氧剥夺(oxygen-glucose deprivation,OGD)6,12,24 hr,以构建PVL体外模型,检测p-ERK蛋白表达变化;给予CG4细胞OGD6,12,24 hr后诱导分化3 days,以构建PVL后分化发育的体外模型,检测p-ERK蛋白表达变化;PD98059干预后给予OGD12,24 hr,检测p-ERK表达的变化。以上蛋白检测均采用蛋白免疫印迹方法。结果与对应时间点的正常对照组比较,OGD实验组p-ERK在OGD6,12,24 hr后均出现明显的下降(P<0.01);OGD后诱导分化3 days实验组,p-ERK在OGD6 hr后诱导分化先出现暂时性的表达升高,随后在12和24 hr时间点,p-ERK表达均出现明显的下降(P<0.01);应用PD98059后OGD12,24 hr实验组,p-ERK反而出现明显的升高(P<0.01)。结论在构建的PVL和随后的分化发育的体外疾病模型中,ERK信号转导通路参与其中。此外,PD98059作为ERK通路的抑制剂在OGD过程中反而起到激活的作用。Objective To study the change of phosphorylated extracellular signal regulated kinase （p-ERK） expression and the effect of ERK inhibitor PD98059 on the periventricular leukomalacia （PVL） model in vitro and discuss whether ERK pathway involves in the pathogenesis of PVL. Methods this study includes three interventional manners： CG4 cells were treated with oxygen-glucose deprivation for 6,12,24 hr to mimic PVL in vitro and the level of p-ERK was analyzed; CG4 cells were treated with oxygen-glucose deprivation for 6,12,24 hr and following by differentiation 3 days to mimic PVL following by the developmental model in vitro and the level of p-ERK was analyzed;CC,4 cells were incubated with PID8059 before OGD 12,24 hr treatment and the level of p-ERK was analyzed. The level of p-ERK expression was analyzed by using western blot. Results Comparing with the nonnal group by which the corresponding time point, the level of p-ERK expression was decreased by OGD 6, 12,24 hr （ P 〈 0. 01 ） ;p-ERK expression was increased by OGD 6 hr following by the differentiation 3 days and then was decreased by OGD 12,24 hr following by the differentiation 3 days （ P 〈 0.01 ） ; Whereas, the level of p-ERK expression was up-regulated by ERK inhibitor with OGD 12,24 hr treatment. Conclusion ERK pathway involves in the pathogenesis of PVL and differentiation model in vitro. Moreover, PD98059, ERK inhibitor, can activate ERK instead of inhibition during OGD.

关 键 词：糖氧剥夺 脑室周围白质软化症 细胞外信号调节激酶 

分 类 号：R742.89[医药卫生—神经病学与精神病学]