水通道蛋白1在培养人眼小梁细胞中的表达及意义

Expression of aquaporin-1 in cultured human trabecular meshwork cells and significance

目的探讨培养人眼小梁细胞水通道蛋白1(AQPl)的存在、定位及意义。方法应用逆转录聚合酶链反应(RT-PCR)检测培养人眼小梁细胞AQP1mRNA的表达。Westernblot用兔抗人AQP1多克隆抗体检测AQP1蛋白表达。免疫荧光测定AQP1在培养人眼小梁细胞的所在部位。结果RT-PCR扩增出一条347bp标志AQP1mRNA的表达产物。Westernblot可见约28000相应位置的发光条带。免疫荧光定位AQP1在培养的人眼小梁细胞胞膜。结论AQP1在培养人眼小梁细胞的细胞膜表达,可能在调节房水通过小梁网流出系统中起到重要作用。

Objective To evaluate the localization and significance of aquaporin-1 ( AQP1) in cultured human trabecular meshwork(TM) cells. Methods The expression of AQP1 mRNA in cultured human TM cells was detected by reverse transcription polymerase chain reaction ( RT-PCR) , and expression of AQP1 protein was determined with rabbit anti-AQP1 polyclonal antibody by Western blot. Immunofluorescence technique was used to localize the expression of AQP1 in cultured human TM cells. Results A 347 bp fragment was amplified by RT-PCR. The band of 28 000 molecular weight could be observed by Western blot. The labeling was selectively localized to the plasma membrane in immunofluorescence. Conclusion AQP1 is expressed in cultured human TM cells plasma membrane,and it may play a physiological role in outflow across TM cells.