PDK1在衰老小鼠小脑中的功能研究  

Functional study of PDK1 in the cerebellum of aging mice

作　　者：彭文婕 齐翠 刘睿 高隽[1,2] Peng Wenjie;Qi Cui;Liu Rui;Gao Jun(Dept of Neurobiology,School of Basic Medicine,Nanjing Medical University,Nanjing 211166;Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases,Shanghai Stomatological Hospital&School of Stomatology,Fudan University,Shanghai 200001)

机构地区：[1]南京医科大学基础医学院神经生物系,南京211166 [2]上海市颅颌面发育与疾病重点实验室,复旦大学附属口腔医院,上海200001

出　　处：《安徽医科大学学报》2024年第12期2065-2071,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:32371053、32100811);江苏省高等学校自然科学研究项目(编号:21KJB180008)。

摘　　要：目的研究3-磷酸肌醇依赖性蛋白激酶1(PDK1)在衰老小鼠小脑浦肯野细胞(PCs)中的功能。方法采用足迹印记实验、高架平衡木实验和转棒实验来检测小鼠步态特征、运动平衡能力和运动协调能力。应用免疫荧光观察小鼠小脑结构和细胞形态学改变。免疫印迹法检测小鼠小脑组织中PDK1、γ型蛋白激酶C(PKCγ)和蛋白激酶B(AKT)蛋白表达水平及核糖体蛋白S6(rpS6)磷酸化水平变化。结果随着年龄的增加,小鼠出现前后足重合程度、运动平衡和运动协调能力降低,小脑体积和PCs胞体体积减少,但PCs密度并未随着小鼠年龄增加而改变;此外,随着年龄的增加,小脑组织中PDK1蛋白表达水平下调,rpS6磷酸化水平、PKCγ和AKT蛋白表达水平也下调。结论随着年龄的增加,小鼠小脑体积及PCs胞体体积减少,这可能与小脑中PDK1表达下降以及rpS6磷酸化、PKCγ和AKT表达水平降低有关,最终影响小脑相关运动调控功能。Objective To investigate the effects of 3-phosphoinositide-dependent protein kinase-1(PDK1)on the purkinje cells(PCs)in aging mice.Methods The footprint test,elevated beam-walk test and rotarod test were used to measure the gait traits,motor balance and motor coordination of mice.Immunoflourescence was used to detect the structure of the cerebellum and the morphology of cells in the cerebellum.The expression level of PDK1,protein kinase C gamma(PKCγ)and protein kinase B(also known as AKT)and the phosphorylation of ribosomal protein S6(rpS6)in the cerebellum were detected by Western blot.Results The mice showed motor defects in the overlap between forelimb and hindlimb,motor balance and motor coordination ability in an age-dependent manner.Furthermore,the cerebellum and the soma size of the PCs decreased in an age-dependent manner.However,the density of the PCs was not affected by age.Additionally,the expression level of the PDK1,PKCγ,AKT and the phosphorylation of rpS6 in the cerebellum were reduced with age.Conclusion The cerebellar size and the cell soma size of PCs decrease in the old mice.These effects may contribute to the reduction of the expression of PKCγand AKT and the phosphorylation of rpS6 in the old mice,ultimately affecting the cerebellar motor coordination.

关 键 词：衰老 浦肯野细胞 3-磷酸肌醇依赖性蛋白激酶1 核糖体蛋白S6 γ型蛋白激酶C 蛋白激酶B 

分 类 号：R339.38[医药卫生—人体生理学]