水孔蛋白1在人腹膜组织的表达及腹膜透析对其表达的影响  

Expression of aquaporin-1 in human peritoneum and effect of peritoneal dialysis on this expression

作　　者：方炜[1] 钱家麒[1] 余志远[1] 陈诗书[2] 

机构地区：[1]上海第二医科大学附属仁济医院肾脏科,200001 [2]上海第二医科大学人类基因治疗研究中心

出　　处：《中华肾脏病杂志》2003年第1期29-33,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金资助项目(39870903);上海市教育委员会资助基金(98QB65)

摘　　要：目的观察水孔蛋白1(aquaporin-1,AQP1)在人腹膜组织的表达以及腹膜透析(腹透)对其表达的影响,以期探讨长期腹透后腹膜超滤功能下降的可能机制。方法采用Western blot、免疫组织化学(组化)以及RT-PCR等技术观察正常对照者、尿毒症非透析患者以及腹透患者的腹膜活检标本AQP1在蛋白质和基因水平表达。结果各组腹膜均有AQP1表达,除了毛细血管和小静脉内皮细胞外,腹膜间皮细胞也表达AQP1。半定量分析表明各组.AQP1蛋白和mRNA的表达量差异均没有显著性意义。结论本研究支持AQP1是腹膜转运超小孔的分子结构。研究结果提示腹膜间皮细胞可能也参与了腹膜跨细胞的水转运。腹透对腹膜AQP1的表达量没有明显影响,因此进一步深入研究AQP1结构或分布的改变,以及与超滤衰竭的关系可能对探讨腹膜超滤衰竭的发生机制具有一定意义。Objective To investigate the expression of aquaporin-1 (AQPl) in human peritoneal tissue and the effect of peritoneal dialysis (PD) on this expression. Methods Peritoneal biopsies were obtained from normal subjects( n = 10), uremic predialysis patients( n = 12) at catheter insertion and PD patients(n = 10) at the time of catheter remove, reinsertion or renal transplantation. Western blot, immunohistochemical staining and RT-PCR techniques were used to investigate AQPl expression in peritoneal tissue. Results All peritoneal samples expressed AQPl at both mRNA and protein levels. Western blot using affinity-purified anti-AQPl antibody revealed a major band at 28 000 as well as more diffuse bands between 35 000 and 50 000. The 28 000 band represents the nonglycosylated form of the protein while 35 000 - 50 000 bands correspond to glycosylated AQPl. Despite the difference in the origin of the tissue, the signal pattern for AQPl was similar in peritoneal samples obtained from normal subjects, uremic predialysis patients or PD patients. When the level of AQPl protein expression in normal subjects was treated as 100%, semi-quantitative analysis showed that peritoneal AQPl protein expression in uremic predialysis patients and PD patients was(84. 0 9±21. 7)% and(92. 0±24. 4)% respectively and there was no significant difference existed. AQPl positive deposits were very specific and restrict distributed in the mesothelial cells, endothelial cells of capillaries, venules and small veins, whereas no signal was detected in arterioles. In some PD patients with disruption of mesothelium, AQPl was not stained in peritoneal lining denuded of mesothelial cells. Within cells, the protein appeared to be located in cell membrane. Semi-quantitative RT-PCR using β-actin as internal standard showed that the levels of AQPl mRNA expression in normal subject, uremic predialysis patients and PD patients were (70. 1 ± 15. 9)% , (59. 3 ± 13. 3)% and (66. 4 ± 12. 9)% respectively, and no significant difference was found in statistical te

关 键 词：水孔蛋白-1 腹膜透析 跨细胞水转运 超滤衰竭 

分 类 号：R459.5[医药卫生—治疗学]