低糖对星形胶质细胞水通道蛋白4表达及分布的影响  

Expression and distribution of AQP4 on astrocytes after glucose deprivation

作　　者：邓江山[1] 赵飞[1] 俞晓燕[1] 赵玉武[1] 

机构地区：[1]上海交通大学附属第六人民医院神经内科,200233

出　　处：《中国神经免疫学和神经病学杂志》2015年第4期263-268,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金资助项目(NO.31271125)

摘　　要：目的探讨低糖体外培养对大鼠原代星形胶质细胞水通道蛋白4(aquaporin 4,AQP4)表达量及分布的影响。方法对体外培养的原代大鼠星形胶质细胞进行0mmol/L(无糖)和1mmol/L葡萄糖低糖培养,以5.5mmol/L葡萄糖作为正常糖浓度对照进行如下实验:(1)采用活细胞成像技术检测无糖培养后的星形胶质细胞的体积变化;(2)以Western blot检测无糖培养0h、3h、6h、12h和24h,以及0mmol/L、1mmol/L和5.5mmol/L葡萄糖培养24h后星形胶质细胞的AQP4表达量;(3)以免疫荧光染色检测0mmol/L、1mmol/L和5.5mmol/L葡萄糖培养6h后AQP4在星形胶质细胞上的分布。结果 (1)与0min时比较,无糖培养15min(1.14±0.03)和30min时(1.15±0.02)星形胶质细胞相对体积均明显增大(P<0.01)。(2)无糖0mmol/L(含血清)培养实验中,与0h(AQP4相对表达量为1)比较,12h(1.55±0.21)、24h时(1.67±0.16)AQP4相对表达量均明显增加(P<0.01);0mmol/L、1mmol/L和5.5mmol/L葡萄糖(含血清)培养24h后,与5.5mmol/L组(AQP4相对表达量为1)比较,0mmol/L组AQP4相对表达量(3.23±0.23)明显增加(P<0.01)。(3)与0h时(AQP4相对表达量为1)比较,无糖(无血清)培养6h后AQP4蛋白相对表达量(1.32±0.09)明显上调(P<0.05),24h时相对表达量(0.80±0.05)下降(P<0.05);0mmol/L、1mmol/L和5.5mmol/L葡萄糖(无血清)培养24h后,与5.5mmol/L(AQP4相对表达量为1)组比较,0mmol/L组AQP4相对表达量(0.75±0.07)明显降低(P<0.05)。无论含或不含血清,与5.5mmol/L葡萄糖组比较,0mmol/L和1mmol/L组AQP4在星形胶质细胞向膜上集中特异性分布更加明显。结论低糖处理可引起星形胶质细胞水肿,AQP4蛋白表达量发生变化和向细胞膜上集中分布。AQP4在低血糖性脑水肿中可能发挥重要作用。Objective To study the effect of glucose deficient media on the expression and distribution of aquaporin 4（AQP4）in the cultured rat primary astrocytes.Methods（1）Primary astrocytes were treated with glucose deficient media（0mmol/L and 1mmol/L）and the control media contained 5.5mmol/L glucose.（2）The cell volume of astrocytes cultured in 0 mmol/L glucose deficient media was measured using live cell imaging technique.The expression of AQP4 was evaluated after 0h,3h,6h,12 hand 24hof culture in media with 0mmol/L glucose or after 24 hof incubation with media containing different concentration of glucose（0mmol/L,1mmol/L and 5.5 mmol/L）by western blot.Distribution of AQP4 on astrocytes cultured in media containing different concentration of glucose（0mmol/L,1mmol/L and 5.5mmol/L）for 6hwas detected by immunofluorescence staining.Results The relative fold of cells volume cultured in media with 0mmol/L glucose were（1.14±0.03）and（1.15±0.02）at 15 min and 30 min respectively,there was a significant increase of the volume compared with that at 0min（P0.01）.In the media containing 0mmol/L glucose（with serum）,the relative fold of protein expression of AQP4were（1.55±0.21）after 12hand（1.67±0.16）after 24 hcompared to that at 0h（the relative expression amount of AQP4 was 1,P0.01）.No significant changes were found after3 hand 6h.After being cultured in media with different concentration of glucose（with serum）for 24 h,the relative fold of AQP4 expression were（3.23±0.23）in the 0mmol/L group（P0.01）and（1.2±0.15）in the1mmol/L group（P0.05）compared with that in the 5.5 mmol/L group（The relative expression amount of AQP4 was 1）.In the media containing 0 mmol/L glucose（The relative expression amount of AQP4 was 1,without serum）,the relative fold of protein expression of AQP4 was（1.32±0.09）after 6 h,there was significant increase compared with that at 0h（P0.05）,and（0.80±0.05）after 24 h,a significant decrease was observed comparing with that at 0h�

关 键 词：水通道蛋白4 低血糖 星形胶质细胞 脑水肿 

分 类 号：R741.02[医药卫生—神经病学与精神病学]