血清对大鼠星形胶质细胞形态和水通道蛋白-4表达的影响  

Effect of Serum on Cultured Astrocytes Morphology and Expression of Aquaporin-4 in Rats

作　　者：贾梅 师忠芳[1] 王玉娇 闫旭[1] 徐立新[1] 董丽萍[1] 李佳欣[1] 陈烨[1] 袁芳[1] JIA Mei;SHI Zhong-fang;WANG Yu-jiao;YAN Xu;XU Li-xin;DONG Li-ping;LI Jia-xin;CHEN Ye;YUAN Fang(Department of Pathophysiology,Beijing Neurosurgical Institute,Beijing Tiantan Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学北京市神经外科研究所病理生理室,首都医科大学附属北京天坛医院,北京市100050

出　　处：《中国康复理论与实践》2018年第4期385-390,共6页Chinese Journal of Rehabilitation Theory and Practice

基　　金：1.国家自然科学基金项目(No.81271286);2.北京市自然科学基金项目(No.7152027);3.北京市神经外科研究所创新探索基金项目(No.2017-5)。

摘　　要：目的通过比较培养大鼠星形胶质细胞在有血清培养基和无血清培养基中的形态及水通道蛋白-4(AQP-4)表达水平,探讨血脑屏障破坏对AQP-4表达的影响。方法取雌性Wistar大鼠大脑皮层细胞,分别在无血清培养基、DMEM培养基加10%胎牛血清(FBS)及无血清培养基加10%FBS中培养。倒置相差显微镜观察星形胶质细胞形态和大小,细胞免疫荧光染色及逆转录实时荧光定量聚合酶链反应(RT-q PCR)检测胶质纤维酸性蛋白(GFAP)、AQP-4及代谢型谷氨酸受体5(m Glu R5)的表达水平。结果在无血清培养基中,星形胶质细胞的细胞核和胞体小且突起细长,折光性强;而在两种添加10%FBS培养基中,星形胶质细胞呈多角扁平形且突起短小,折光性弱。细胞免疫荧光及RT-q PCR结果分析显示,两种加10%FBS培养基中星形胶质细胞GFAP和AQP-4的蛋白和m RNA表达水平均低于无血清培养基(P<0.001),而m Glu R5蛋白和m RNA表达无显著性差异(P>0.05)。结论血清改变培养星形胶质细胞的形态,降低GFAP和AQP-4的表达。Objective To explore the effect of blood-brain barrier disruption on expression of AQP-4,through comparing the cell morphology and the expression of aquaporin-4(AQP-4)of cultured astrocytes in medium with and without fetal bovine serum(FBS).Methods Cerebral cortical astrocytes from female Wistar rats were cultured in serum free medium,DMEM supplemented with 10%FBS,and serum free medium supplemented with 10%FBS.Phase contrast microscope was used to detect the cell morphology and cell size.Immunofluorescence staining and reverse real-time quantitative polymerase chain reaction(RT-qPCR)were used to examine the expression of glial fibrillary acidic protein(GFAP),AQP-4 and metabotropic glutamate receptor 5(mGluR5).Results Astrocytes in serum free medium showed extensive process bearing morphology,small body and nucleus,and high refractivity.In contrast,in two kinds of 10%FBS-containing medium,astrocytes were flat with large body and nucleus,weak refractivity,as well as short process.Analysis of immunofluorescence staining and RT-qPCR revealed a down-regulation of GFAP and AQP-4 protein and mRNA expression in two kinds of 10%FBS-containing medium,compared with that in serum free medium(P<0.001),however,there was no difference in mGluR5 protein and mRNA expression(P>0.05).Conclusion FBS changed astrocyte morphology and down-regulated the expression of GFAP and AQP-4.

关 键 词：星形胶质细胞 血清 水通道蛋白-4 胶质纤维酸性蛋白 代谢型谷氨酸受体5 大鼠 

分 类 号：R651.1[医药卫生—外科学]