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目的:通过观察重复性低氧刺激对蛋白激酶CβI和βII亚型(cPKCβI,cPKCβII)膜转位水平(或激活程度)的影响,初步探讨cPKCs特定亚型在脑低氧预适应发生发展过程中的作用。方法:按我室已建立的小鼠低氧预适应模型方法,制备重复性低氧1-4次小鼠(H1-H4)。应用SDS-聚丙烯酰胺凝胶蛋白电泳(SDS-PAGE)、蛋白印迹(Western blot)等生化技术,并结合应用Gel Doc凝胶成像系统,半定量检测小鼠海马和大脑皮层组织内cPKCβI和βII的膜转位水平。结果:随低氧刺激次数(H1-H4)的增加,小鼠海马组织内cPKCβII的膜转位水平升高,H4组膜转位水平显著高于H0(100%,n=10)和H1(79·5%±10·7%,n=10)组(173·3%±21·3%,P<0·01,n=10);同样,大脑皮层内cPKCβII膜转位水平也随低氧次数的增加而升高,H3(119·8%±7·7%,n=6)和H4(131·8%±4·7%,n=6)组膜转位水平均显著高于H0(100%,n=6)组(P<0·05,n=6);而cPKCβI亚型的膜转位水平无论在大脑皮层还是海马组织内的变化均不显著(P>0·05,n=8)。结论:cPKCβII膜转位(激活)可能在脑低氧预适应的发生发展过程中发挥着重要作用。
OBJECTIVE: To investigate the effect of repetitive hypoxia on the translocation (or activation) of protein kinase CβI and βII subtypes (cPKCβI, cPKCβII), and to investigate the role of specific subtypes of cPKCs in the development of hypoxic preconditioning Role. Methods: Hypoxic preconditioning mice model established in our laboratory was used to prepare repeated hypoxia 1-4 mice (H1-H4). The biochemical techniques such as SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot were used to detect the expression of cPKCβI and βII in the hippocampus and cerebral cortex of mice by using Gel Doc imaging system. Membrane translocation level. Results: The membrane translocation level of cPKCβII in hippocampus of mice increased with the increase of the number of hypoxia stimulation (H1-H4). The level of membrane translocation in H4 group was significantly higher than that of H0 (100%, n = 10) and H1 79.5% ± 10.7%, n = 10) (173.3% ± 21.3%, P <0.01, n = 10). Similarly, the level of cPKCβII translocation in the cerebral cortex also decreased The translocation levels of the three groups of H3 (119.8% ± 7.7.7%, n = 6) and H4 (131.8% ± 4.77%, n = 6) H0 (100%, n = 6) group (P <0 · 05, n = 6). However, the cPKCβI subtypes had no significant change in the level of membrane translocation in cerebral cortex and hippocampus (P> 0.05) , n = 8). Conclusion: cPKCβII membrane translocation (activation) may play an important role in the development of hypoxic preconditioning.