Hodeify Rawad F, Kreydiyyeh Sawsan Ibrahim
Department of Biology, Faculty of Arts & Sciences, American University of Beirut, Beirut, Lebanon.
Prostaglandins Leukot Essent Fatty Acids. 2007 May;76(5):285-92. doi: 10.1016/j.plefa.2007.02.002. Epub 2007 May 3.
An effect of PGE2 on water and chloride absorption was already established in a previous work. This study is an attempt to find the mechanism of action of the prostaglandin by investigating the involvement of three major transporters namely the Na+ -K+ ATPase, the Na+/H+ exchanger and the Na+ K+ 2Cl- cotransporter. Rats were injected with PGE2 and 15 min later, the colon was perfused in situ with Krebs Ringer buffer, and net water and chloride absorption were determined. When the involvement of the cotransporter and/or the exchanger was investigated, animals were injected with, respectively, furosemide and amiloride 10 min before PGE2. Superficial and crypt colonocytes were then isolated and the protein expression of the Na+ -K+ ATPase and the Na+ K+ 2Cl- was determined by western blot analysis. The effect of PGE2 on the pump activity in presence or absence of the transporters' inhibitors was also studied. PGE2 decreased net water and chloride absorption from the colon, increased the Na+ -K+ ATPase activity in superficial cells and reduced it in crypt cells. The prostaglandin was found to stimulate secretion in superficial cells by targeting the Na+ K+ 2Cl- symporter, and reduce absorption in crypt cells by targeting the Na+/H+ antiporter. Changes in the activity of the pump are secondary to changes in the activity of the other transporters.
PGE2对水和氯离子吸收的影响在之前的一项研究中已经得到证实。本研究试图通过研究三种主要转运蛋白,即Na+-K+ATP酶、Na+/H+交换体和Na+K+2Cl-协同转运体的参与情况,来探寻前列腺素的作用机制。给大鼠注射PGE2,15分钟后,用Krebs-Ringer缓冲液原位灌注结肠,测定水和氯离子的净吸收量。当研究协同转运体和/或交换体的参与情况时,在注射PGE2前10分钟分别给动物注射速尿和阿米洛利。然后分离表层和隐窝结肠细胞,通过蛋白质免疫印迹分析测定Na+-K+ATP酶和Na+K+2Cl-转运体的蛋白表达。还研究了在存在或不存在转运体抑制剂的情况下PGE2对泵活性的影响。PGE2降低了结肠水和氯离子的净吸收,增加了表层细胞中Na+-K+ATP酶的活性,降低了隐窝细胞中该酶的活性。发现前列腺素通过作用于Na+K+2Cl-同向转运体刺激表层细胞分泌,并通过作用于Na+/H+反向转运体减少隐窝细胞的吸收。泵活性的变化是其他转运体活性变化的继发结果。