低渗液对AQP4蛋白在体外培养星形胶质细胞中的表达变化  

Changes of astrocytes AQP4 protein expression during hypotonic medium in vitro

作　　者：李燕华[1] 孙善全[1] 

机构地区：[1]重庆医科大学神经生物学研究室,重庆400016

出　　处：《中国急救医学》2004年第2期87-90,共4页Chinese Journal of Critical Care Medicine

基　　金：国家自然科学基金资助课题 (No 3 0 0 70 2 47)

摘　　要：目的　研究低渗培养液对星形胶质细胞AQP4蛋白的表达变化 ,并探讨其在脑水肿中的作用机制。方法　取出生后 2d的Wistar大鼠大脑皮层进行星形胶质细胞纯培养 ,分别予不同程度的低渗培养液作用于星形胶质细胞 ,建立星形胶质细胞对低渗压反应的实验模型。应用四甲基偶氮唑蓝 (MTT)比色法测定细胞活性 ,采用免疫细胞化学法检测AQP4蛋白的表达 ,通过光镜观察低渗液对星形胶质细胞形态学影响。结果　体外培养的星形胶质细胞在不同程度的低渗培养液作用 3、6、12、2 4h后 ,各时相点的AQP4蛋白表达水平均明显高于对照组 (P <0 .0 5 ) ,而且与作用时间和低渗压的程度密切相关 ;在光镜下 ,星形胶质细胞出现典型的细胞水肿形态学改变 ,其活性明显下降。结论　低渗液可导致星形胶质细胞水肿 ,AQP4蛋白表达增强 ,而且AQP4表达上调与低渗压的程度和作用时间有关 ,抑制AQP4的表达可能为脑水肿的治疗提供新的理论依据。Objective To study the expression of aquaporin-4(AQP4) protein in cultured rat astrocytes after exposure to hypotonic medium and the mechanisms of brain edema formation.Methods Rat cerebral cortical astrocytes from 2 days newborn Wistar rat were undergone the pure culture and its model of cell edema was established by exposure to hypotonic medium. The reactivity and expression of AQP4 protein of astrocytes to hypotonic medium were studied by morphological changes, immunocytochemistry, MTT and imaging analysis. Results The OD values (MTT) of astrocytes cultured for 3, 6, 12, 24h with hypotonic medium(268, 254, 240 mOsm) were significantly lower than those in the control group(P<0.05). Microscope showed typical features of cell edema. Immunocytochemistry analysis confirmed that the expression of AQP4 protein was markedly increased in comparison with that of the control group(P<0.05). Conclusion Hypotonic medium could induce the expression of AQP4 protein. Upgraduated expression of AQP4 is correlated with hypotonic medium and time dependent manner. AQP4 inhibition may be a new direction for potential therapeutic intervention in brain edema.

关 键 词：星形胶质细胞 低渗培养液 水通道蛋白-4 体外培养 

分 类 号：R741[医药卫生—神经病学与精神病学]