低氧预适应小鼠脑海马组织内与cPKCγ相互作用的ERK1/2磷酸化水平降低

Decreased phosphorylation of cPKCγ-interacting ERK1/2 in the hippocampus of hypoxic preconditioned mice

目的探讨与经典型蛋白激酶Cγ(cPKCγ)相互作用的细胞外信号调节激酶1/2(ERK1/2)的磷酸化水平在小鼠脑低氧预适应形成过程中的变化。方法用成年雄性BalB/c小鼠制备整体低氧预适应模型,将小鼠随机分为正常对照(H0)、早期低氧预适应(H3)和延迟性低氧预适应(H6)3组。应用免疫共沉淀,Western blot等方法检测胞质和膜相关成分中与cPKCγ相互作用的ERK1/2的磷酸化水平。结果小鼠海马组织中ERK1/2能与cPKCγ免疫共沉淀,且与H0组相比,H3及H6组ERK1/2的204位酪氨酸磷酸化水平明显降低(P<0.05)。结论小鼠海马组织中,cPKCγ与ERK1/2存在相互作用,且可能参与了脑低氧预适应的发生和发展。

Objective To determine the changes in phosphorylation of conventional protein kinase Cγ（cPKCγ）- interacting extracellular signal regulated kinase 1/2 （ERK1/2） in the hippocampus of hypoxic preconditioned （HPC） mice. Methods Healthy male BalB/c mice were used to develop ＂auto-hypoxia＂-induced HPC mice and randomly divided into 3 groups as follows： normoxic control （ H0）, early （ H3 ） and delayed hypoxic preconditioned groups （H6）. Co-immunoprecipitation and Western blot were applied to quantitatively analyze the phosphorylation level of cPKCγ-interacting ERK1/2 in cytosolic and particulate fractions of hippocampus of HPC mice. Results ERK1/2 was co-immunoprecipitated by cPKCγ/in hippocampus of mice. The phosphorylation level of cPKCγ/interacting ERK1/2 at tyrosine 204 decreased significantly in the hippocampus of H3 and H6 groups as compared with that of the H0 group （P 〈 0.05 ）. Conclusion cPKCγ interacts with ERK1/2 in the hippocampus of mice, and their interaction might be involved in the development of cerebral hypoxic preconditioning of mice.