摘要
目的观察重复高压氧预处理对大鼠脑皮层神经元缺氧损伤中丝裂原活化蛋白激酶p38(p38 MAPK)与其活化状态P-p38 MAPK蛋白表达水平的影响。方法原代培养的大鼠脑皮层神经元分为4组:正常对照组(Con组),细胞置常规培养箱内培养;高压氧预处理组(P组),培养细胞行0.35 MPa、2 h高压氧预处理,1次/d,连续3 d;缺氧处理组(H组),细胞置三气培养箱内培养4 h,设置培养条件为95%N2、5%CO2;高压氧预处理加缺氧处理组(PH组),高压氧预处理后行缺氧处理,条件设置同上,每项实验操作3次。四组细胞均在处理4 h后行免疫细胞化学染色和免疫蛋白印迹(Western blot)检测。实验数据采用单因素方差分析进行统计。结果 p38 MAPK表达水平在各组之间差异无统计学意义;缺氧损伤后,H组P-p38 MAPK水平比Con组高(q=8.15,P<0.01);高压氧预处理使PH组神经元P-p38 MAPK水平比H组低(q=4.06,P<0.05)。结论重复高压氧预处理抑制p38 MAPK激活,可能参与了对皮层神经元缺氧损伤的保护作用。
Objective To investigate the effect of repeated hyperbaric oxygen preconditioning on p38 mitogen-activated protein kinases （p38 MAPK） and phosphorylation p38 mitogen-aetivated protein kinases（ P-p38 MAPK） in hypoxia injury of rat cortical neurons. Methods Primary cultured rat cortical neurons were randomly assigned to 4 groups： control group （Con group ）, general cultured in common incubator; hyperbaric oxygen preconditioning group （P group ）, preconditioning with hyperbaric oxygen in proposal of 0. 35 MPa, 2 h, once/d, 3 d consecutively; hypoxia group （ H group）, cortical neurons were cultured in three gas incubator, set the culture condition as 95% N2, 5% CO2; hyperbaric oxygen preconditioning ＋ hypoxia group （PH group ）, cortical neurons were cultured in three gas incubator after preconditioning with hyperbaric oxygen, the culture condition was same as H group. Immunocytochemical stain and Western blot were performed after 4 h in hypoxia culture. Data were analyzed by one-factor analysis of variance. Results The level of p38 MAPK in cortical neurons among four groups had no significant difference. Compared with Con group, the level of P-p38 MAPK in H group increased evidently after hypoxia injury（ q = 8.15,P 〈 0. 01 ）, however, the P-p38 MAPK level of PH group decreased significantly compared with that of H group（ q = 4.06,P 〈 0. 05 ）. Conclusion The protective effect of repeated hyperbaric oxygen preconditioning on hypoxia injury in cortical neurons of rat may be involved in the suppressed activation of p38 MAPK.
出处
《中华脑科疾病与康复杂志（电子版）》
2012年第6期24-27,共4页
Chinese Journal of Brain Diseases and Rehabilitation（Electronic Edition）
基金
国家自然科学基金(81000564)
关键词
高压氧
神经元
预处理
缺氧
脑
丝裂原活化蛋白激酶磷酸酶类
Hyperbaric oxygen
Neurons
Preconditioning
Hypoxia, brain
Mitogenactivated protein kinase phosphatases
