Matthews J B, Hassan I, Meng S, Archer S Y, Hrnjez B J, Hodin R A
Division of General Surgery, Department of Surgery, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.
J Clin Invest. 1998 May 15;101(10):2072-9. doi: 10.1172/JCI1042.
The basolateral Na-K-2Cl cotransporter (NKCC1) is a key component of the intestinal crypt cell secretory apparatus. Its fate during the transition to absorptive enterocyte and the potential impact of its altered expression on secretory output have not been addressed. In this report, NKCC1 mRNA was found to be expressed in rat jejunal crypt but not villus cells. Butyrate treatment of intestinal epithelial HT29 cells induced a differentiation pattern that recapitulated the rat intestinal crypt-villus axis, with NKCC1 mRNA levels decreasing in a time- and dose-dependent fashion in parallel with upregulation of apical brush-border markers. Butyrate but not acetate or proprionate decreased basal and cAMP-stimulated bumetanide-sensitive K+ (86Rb) uptake in both HT29 cells and the Cl--secreting T84 line. Butyrate markedly decreased transepithelial Cl- secretion in confluent T84 monolayers without effect on cAMP-regulated apical Cl- efflux. We conclude that NKCC1 regulation during enterocyte differentiation occurs at the level of gene expression, and that selective downregulation of NKCC1 gene expression and function by butyrate leads to a profound decrease in transepithelial Cl- secretion. These data emphasize the importance of NKCC1 in determining epithelial secretory capacity and suggest the possibility of modulation of the enterocytic transport phenotype as therapy for diarrheal disorders.
基底外侧钠-钾-2氯协同转运蛋白(NKCC1)是肠隐窝细胞分泌装置的关键组成部分。在向吸收性肠上皮细胞转变过程中它的命运以及其表达改变对分泌输出的潜在影响尚未得到研究。在本报告中,发现NKCC1 mRNA在大鼠空肠隐窝中表达,但在绒毛细胞中不表达。用丁酸盐处理肠上皮HT29细胞可诱导出一种重现大鼠肠隐窝-绒毛轴的分化模式,其中NKCC1 mRNA水平随时间和剂量依赖性降低,同时顶端刷状缘标记物上调。丁酸盐而非乙酸盐或丙酸盐可降低HT29细胞和分泌氯离子的T84细胞系中基础的和cAMP刺激的布美他尼敏感的钾离子(86Rb)摄取。丁酸盐显著降低汇合的T84单层细胞的跨上皮氯离子分泌,而对cAMP调节的顶端氯离子外流无影响。我们得出结论,肠上皮细胞分化过程中NKCC1的调节发生在基因表达水平,并且丁酸盐对NKCC1基因表达和功能的选择性下调导致跨上皮氯离子分泌显著减少。这些数据强调了NKCC1在决定上皮分泌能力方面的重要性,并提示调节肠细胞转运表型作为腹泻性疾病治疗方法的可能性。