体外培养大鼠星形胶质细胞水通道蛋白4的表达  

Expression of Aquaporin 4 in Astrocytes of Rats in Vitro

作　　者：徐立新 董丽萍 师忠芳 闫旭[1,2] 杨少华 袁芳[1,2] XU Li-xin;DONG Li-ping;SHI Zhong-fang;YAN Xu;YANG Shao-hua;YUAN Fang(Department of Pathophysiology, Beijing Neurosurgical Institute, Beijing 100050, China;Beijing Tiantan Hospital Capital Medical University, Beijing 100050, China;Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, FortWorth, Texas 76107, USA)

机构地区：[1]北京市神经外科研究所病理生理室,北京市100050 [2]首都医科大学附属北京天坛医院,北京市100050 [3]北德克萨斯大学医学中心神经与药理系,美国德克萨斯州沃思堡76107

出　　处：《中国康复理论与实践》2017年第9期1051-1055,共5页Chinese Journal of Rehabilitation Theory and Practice

基　　金：国家自然科学基金项目(No.81271286);北京市自然科学基金项目(No.7152027)

摘　　要：目的研究水通道蛋白4(AQP4)在体外培养大鼠星形胶质细胞中的表达规律。方法取新生1 d Wistar大鼠大脑皮层行星形胶质细胞原代培养,分别于原代培养3 d、5 d、7 d、9 d及传代培养9 d行胶质纤维酸性蛋白(GFAP)免疫荧光染色;实时荧光定量聚合酶链反应及AQP4免疫荧光染色检测AQP4 m RNA及蛋白的表达。结果原代及传代培养不同时间点95%以上细胞GFAP免疫荧光染色阳性,且GFAP表达量逐渐增加。原代培养3 d有AQP4 m RNA表达,3 d和5 d AQP4 m RNA表达水平无显著性差异(P>0.05),7 d AQP4 m RNA表达升高(P<0.05),9 d持续升高(P<0.05);传代培养9 d与原代培养9 d AQP4 m RNA表达无显著性差异(P>0.05)。AQP4蛋白表达与AQP4 m RNA表达一致。结论大鼠星形胶质细胞原代培养3 d即有AQP4表达,之后逐渐增加,9 d左右表达渐趋稳定。Objective To explore the expression of aquaporin4(AQP4)in primary and secondary cultured rat astrocytes.Methods Rat cortical astrocytes from a newborn(one day)Wistar rat were cultured.Astrocytes were identified with immunofluorescence staining of glial fibrilillary acidic protein(GFAP).The expression of AQP4was determined with real-time quantitative polymerase chain reaction and immunofluorescence staining three,five,seven and nine days of primary culture,and nine days of secondary culture.Results The purity of GFAP-positive cells was more than95%.The expression of AQP4mRNA was found three days of primary culture,remained unchanged five days of primary culture(P>0.05),and increased seven and nine days of primary culture(P<0.05).The expression of AQP4mRNA was not different between nine days of primary culture and nine days of secondary culture(P>0.05).AQP4immunofluorescence staining showed the same trend of AQP4mRNA.Conclusion AQP4may express since three days of primary culture in rat astrocytes in vitro,and increase slowly until nine days of primary culture.

关 键 词：水通道蛋白4 星形胶质细胞 细胞培养 大鼠 

分 类 号：R338.2[医药卫生—人体生理学]