雌激素对H_2O_2诱导PC12细胞氧化应激的保护作用研究  

A study on the Neuroprotective effect of estrogen on H_2O_2-induced oxidative stress in PC12 cells

作　　者：裴新[1] 陆地[2] 周宇冠 陈穗云[1] 

机构地区：[1]云南大学生命科学学院,云南昆明650091 [2]昆明医学院,云南昆明650031 [3]禄劝县第二人民医院,云南禄劝651506

出　　处：《云南大学学报（自然科学版）》2010年第5期608-612,共5页Journal of Yunnan University(Natural Sciences Edition)

基　　金：国家自然学基金资助项目(30560170);云南省教育厅科学研究基金重点项目资助(07Z10392)

摘　　要：为了探讨雌激素对H2O2诱导PC12细胞氧化损伤的影响及其可能机制.用H2O2处理PC12细胞建立氧化应激模型,并加入雌激素预处理作为保护.MTT法检测细胞活力,利用荧光探针对细胞内ROS进行荧光染色检测荧光强度,Hoechst/PI荧光染色,分析细胞凋亡情况,Western blotting检测MAPK(Mitogen-Activated Protein Kinase)的磷酸化水平.结果显示:H2O2作用于PC12细胞后显著降低细胞活力,10-8～10-6mol/L的雌激素预处理后均能部分阻断H2O2对细胞活力的影响.H2O2能显著增强PC12细胞的ROS荧光强度,雌激素可明显减少H2O2处理后PC12细胞内的ROS含量.H2O2激活MAPK,而雌激素抑制了p38的磷酸化,增加了ERK的磷酸化.以上结果表明,雌激素可以拮抗H2O2诱导的氧化应激,抑制p38的磷酸化,增强ERK的磷酸化可能是其发挥保护作用的机制之一.To investigate the neuroprotective effect of estrogen on the H2O2-induced oxidative stress in PC12 cells,oxidative damaged PC12 cells was treated by H2O2 to construct the model on the oxidative stress reaction,and estrogen was added as a pretreatment to protect the cells.The survival rate of the cells was determined by MTT method,intracellular ROS generation was detected by DCFH-DA fluorescent probe,apoptosis was analyzed by Hoechst/PI fluorescence staining,and the level of mitogen-activated protein（MAP） kinase phosphorylation was detected by the Western blotting.The results showed that：The pretreated pheochromocytoma cell line PC12 by estrogen（0.001—1 μmol/L） recovered significantly from the hydrogen peroxide（H2O2）-induced cell injury.The intensity of DCFH-DA fluorescence was significantly increased after the treatment by H2O2,indicating that ROS generation was successfully induced in PC12 cell,and estrogen（1μmol/L） decreased the DCFH-DA fluorescence intensity significantly.Estrogen suppressed H2O2-induced phosphorylation of p38MAPK and increased H2O2-induced phosphorylation of ERK1 /2.It was suggest that estrogen protected against H2O2-induced oxidative stress in vitro and its protective effect was associated with the inhibition of p38 MAPK and the activation of ERK.

关 键 词：雌激素 氧化应激 丝裂原活化蛋白激酶 

分 类 号：Q591.8[生物学—生物化学]