体外培养大鼠星形胶质细胞水通道蛋白4的表达

doi:10.3969/j.issn.1006-9771.2017.09.012

《中国康复理论与实践》 ›› 2017, Vol. 23 ›› Issue (9): 1051-1055.doi: 10.3969/j.issn.1006-9771.2017.09.012

体外培养大鼠星形胶质细胞水通道蛋白4的表达

徐立新^{1, 2}, 董丽萍^{1, 2}, 师忠芳^{1, 2}, 闫旭^{1, 2}, 杨少华³, 袁芳^{1, 2}

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。

收稿日期:2017-05-17 修回日期:2017-06-22 出版日期:2017-09-25 发布日期:2017-10-10
通讯作者: 董丽萍,女,研究员。E-mail: d08270827@163.com。
作者简介:徐立新(1966-),女,汉族,北京市人,主管技师,主要研究方向：神经系统相关疾病基础研究。
基金资助:
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)

Expression of Aquaporin 4 in Astrocytes of Rats in Vitro

XU Li-xin^{1, 2}, DONG Li-ping^{1, 2}, SHI Zhong-fang^{1, 2}, YAN Xu^{1, 2}, YANG Shao-hua³, YUAN Fang^{1, 2}

1. Department of Pathophysiology, Beijing Neurosurgical Institute, Beijing 100050, China;
2. Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China;
3. Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, Texas 76107, USA

Received:2017-05-17 Revised:2017-06-22 Published:2017-09-25 Online:2017-10-10
Contact: Correspondence to DONG Li-ping. E-mail: d08270827@163.com

摘要/Abstract

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 mRNA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 mRNA表达,3 d和5 d AQP4 mRNA表达水平无显著性差异(P>0.05),7 d AQP4 mRNA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 mRNA表达无显著性差异(P>0.05)。AQP4蛋白表达与AQP4 mRNA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。

关键词: 水通道蛋白4, 星形胶质细胞, 细胞培养, 大鼠

Abstract: ObjectiveTo explore the expression of aquaporin 4 (AQP4) in primary and secondary cultured rat astrocytes. MethodsRat 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 AQP4 was 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. ResultsThe purity of GFAP-positive cells was more than 95%. The expression of AQP4 mRNA 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 AQP4 mRNA was not different between nine days of primary culture and nine days of secondary culture (P>0.05). AQP4 immunofluorescence staining showed the same trend of AQP4 mRNA. ConclusionAQP4 may express since three days of primary culture in rat astrocytes in vitro, and increase slowly until nine days of primary culture.

Key words: aquaporin 4, astrocyte, cell culture, rats

中图分类号:

R338.2

徐立新, 董丽萍, 师忠芳, 闫旭, 杨少华, 袁芳. 体外培养大鼠星形胶质细胞水通道蛋白4的表达[J]. 《中国康复理论与实践》, 2017, 23(9): 1051-1055.

XU Li-xin, DONG Li-ping, SHI Zhong-fang, YAN Xu, YANG Shao-hua, YUAN Fang. Expression of Aquaporin 4 in Astrocytes of Rats in Vitro[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2017, 23(9): 1051-1055.

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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

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