摘要
目的:检测急性重复低氧预适应小鼠海马脑组织中蛋白磷酸酶(protein phosphatase,pp)1γ的变化,探讨pp1γ的变化在低氧预适应脑保护中的作用。方法:成年昆明小鼠随机分成低氧暴露1次组(H1)或低氧暴露4次(H4)组和常氧组(H0)。低氧结束后取海马组织,应用Real-time PCR技术和蛋白印记技术检测pp1γ表达的变化;应用磷酸酶活性实验对pp1活性的变化进行检测;应用甲基化测序实验对pp1γ启动子区(95~321 bp)甲基化变化进行检测。结果:pp1γ的mRNA在H4组表达较H0组降低,差异有统计学意义(P<0.01);pp1γ的蛋白质变化与mRNA变化一致,在H4组的表达较H0组降低,差异有统计学意义(P<0.05);pp1γ水解磷酸的活性在H4组和H1组均低于H0组(P<0.05);pp1γ启动子区(95~321 bp)DNA甲基化情况在三组间未见差异。结论:pp1γ变化可能参与了低氧预适应小鼠获得的脑保护,但该变化可能与其启动子区(95 bp^321 bp)的甲基化无关。
Objective：To study the changes of protein phosphatase（ pp） 1γ in hippocampus of mice with acute repeated hypoxic exposure and to explore the neuroprotection of pp1γ in the process of hypoxia preconditioning.Methods：Mice were randomly divided into 3 groups：1 run of hypoxia exposure（ H1）,4 runs of hypoxia exposure（ H4） and no hypoxia exposure group（ control group,H0）.Real-time PCR and Western Blot were used to examine the expression change of pp1γ in hippocampus after treatment.The protein phosphatase activity assay was used to examine the change of pp1 activity.Bisulfite-Modified DNA Sequencing was used to detect the DNA methylation of pp1γ promoter（ 95 - 321 bp）.Results：The mRNA level of pp1γ was found to be significantly decreased in H4 compared with H0（ P〈0.01）.The change of ppr at protein level was similar to that of mRNA,being significantly decreased in H4 compared with H0（ P〈0.05）.The activity of pp1γ was sinificantly decreased in H4 and H1 groups,less active than that in H1（ P〈0.05）.While DNA methylation of the promoter of PP1γ（ 95 - 321 bp） showed no change in the three groups.Conclusion：The changes of pp1γ may be involved in protection of the brain in hypoxic preconditioned mice,which may not be related to its promoter（ 95 bp - 321 bp） DNA methylation change.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2014年第6期644-648,共5页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金(81060212
81160244
81360316
81460283)
内蒙古自然科学基金资助项目(2010BS1104
2014MS0810)
中国博士后基金项目(20080430851)
内蒙古自治区高等学校科研项目(NJZY12221
NJZY12225)
内蒙古自治区高等学校青年科技英才支持计划资助(NJYT-13-A10)
研究生科研创新项目(S201410127(Y02))
教育部留学回国人员科研启动基金(46批)
关键词
缺氧预适应
海马
蛋白磷酸酶1γ
小鼠
hypoxic preconditioning
hippocampus
protein phosphatase 1γ
mouse
