目的探讨糖尿病脑病大鼠皮层中蛋白激酶Cγ(PKCγ)表达及作用。方法36只成年雄性SD大鼠随机分为正常对照组(N组)、2型糖尿病模型组(DM组)、2型糖尿病模型+侧脑室DMSO注射组(DM+DMSO组)和2型糖尿病模型+侧脑室注射G06983组(DM+G06983组),每组各9只。N组采用正常饲料喂养,DM组、DM+DMSO组、DM+G06983组通过高脂饮食和腹腔注射链尿佐菌素(STZ)建立2型糖尿病模型,之后DM+DMSO组、DM+G06983组分别予以侧脑室注射DMSO、G069835uL。通过Morris水迷宫检测大鼠空间记忆能力。放免法检测血清胰岛素水平。取大鼠前额叶皮层组织,通过Weaernblot法检测PKCγ蛋白表达和亚细胞分布。结果与N组比较,DM组大鼠水迷宫逃避潜伏期时间延长,搜索策略能力降低,空腹血糖显著升高,血清胰岛素水平和体重明显减少,差异均有统计学意义(P〈0.05)。与DM组、DM+DMSO组比较,DM+G06983组大鼠水迷宫逃避潜伏期进一步延长,搜索策略得分显著降低,差异均有统计学意义(P〈0.05)。与N组比较,DM组大鼠皮层PKCγ蛋白全细胞表达量未有明显变化(P〉0.05),膜转位水平明显高于正常脑组织,差异有统计学意义(P〈0.05)。结论PKCγ膜转位(激活)可能在2型糖尿病脑病的发展过程中起到神经保护作用。
Objective To explore the expression and role of PKCγ in the cortex of diabetic encephalopathy of rats. Methods 36 adult male SD rats were randomly divided into four groups (9 rats in each group): normal control group (N group), type 2 diabetes model group (DM group), type 2 diabetes model + DMSO intracerebroventricular injection group (DM+DMSO group) and type 2 diabetes model+Go6983 intraeerebroventricular injection group (DM+Go6983 group). The rats in N group were fed with normal feedstuff, the models of DM, DM+DMSO and DM+Go6983 groups were established by application of high-fat diet and intraperitoneal injection of streptozocin (STZ). 5 μL DMSO or Go6983 was intracerebroventricular injected in DM +DMSO group or DM+Go6983 group respectively. The spatial memories were determined by Morris water maze test in rats. The serum insulin levels were detected by radio-immunity method. PKCγ protein expression and subcellular distribution in the prefrontal cortex of rats were explored by Western blot method. Results Compared with N group, the escape latencies of water maze in DM group were longer, the scores of search platform strategy in Morris water maze test of DM group were lower, the fasting plasma glucose of DM group increased significantly, while serum insulin levels and body weight decreased remarkably, the differences were statistically significant (P 〈 0.05). Compared with DM group and DM+DMSO group, the water maze latency of rats in DM+Go6983 group extended and the scores of search platform strategy reduced, the differences were statistically significant (P 〈 0.05). Compared with N group, the expression of PKCγprotein in cortex of DM group had no change (P 〉 0.05). The mere-brane translocation of PKCγ was significantly higher than that of normal brain tissues, the difference was statistically significant (P 〈 0.05). Conclusion The PKCγ membrane translocation (activated) may play a neuroprotective role in the development of type 2 di