Moretó M, Cristià E, Pérez-Bosque A, Afzal-Ahmed I, Amat C, Naftalin R J
Departament de Fisiologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain.
J Membr Biol. 2005 Jul;206(1):43-51. doi: 10.1007/s00232-005-0772-5.
Fluid and electrolyte absorption by colonic crypts depends on the transport properties of crypt cellular and paracellular routes and of the pericryptal sheath. As a low-Na(+) diet increases aldosterone and angiotensin II secretion, either hormone could affect absorption. Control and adrenalectomized (ADX) Sprague-Dawley rats were kept at a high-NaCl (HS) diet and then switched to low-NaCl (LS) diet for 3 days. Aldosterone or angiotensin II plasma concentrations were maintained using implanted osmotic mini-pumps. The extracellular Na(+) concentration in isolated rat distal colonic mucosa was determined by confocal microscopy using a low-affinity Na(+) -sensitive fluorescent dye (Sodium red, and Na(+) -insensitive BODIPY) bound to polystyrene beads. Crypt permeability to FITC-labelled dextran (10 kDa) was monitored by its rate of escape from the crypt lumen into the pericryptal space. Mucosal ion permeability was estimated by transepithelial electrical resistance (TER) and amiloride-sensitive short-circuit current (SCC). The epithelial Na(+) channel, ENaC, was determined by immunolocalization. LS diet decreased crypt wall permeability to dextran by 10-fold and doubled TER. Following ADX, aldosterone decreased crypt wall dextran permeability, increased TER, increased Na(+) accumulation in the pericryptal sheath and ENaC expression even in HS. Infusion of angiotensin II to ADX rats did not reverse the effects of aldosterone deprivation. These findings indicate that aldosterone alone is responsible for both the increase in Na(+) absorption and the decreased paracellular and pericryptal sheath permeability.
结肠隐窝对液体和电解质的吸收取决于隐窝细胞途径、细胞旁途径以及隐窝周围鞘的转运特性。由于低钠饮食会增加醛固酮和血管紧张素II的分泌,这两种激素都可能影响吸收。将对照大鼠和肾上腺切除(ADX)的Sprague-Dawley大鼠维持在高氯化钠(HS)饮食,然后切换到低氯化钠(LS)饮食3天。使用植入式渗透微型泵维持醛固酮或血管紧张素II的血浆浓度。通过共聚焦显微镜,使用与聚苯乙烯珠结合的低亲和力钠敏感荧光染料(钠红和钠不敏感的硼二吡咯)来测定分离的大鼠远端结肠黏膜中的细胞外钠浓度。通过FITC标记的葡聚糖(10 kDa)从隐窝腔逃逸到隐窝周围间隙的速率来监测隐窝对其的通透性。通过跨上皮电阻(TER)和氨氯地平敏感的短路电流(SCC)来估计黏膜离子通透性。通过免疫定位来确定上皮钠通道ENaC。LS饮食使隐窝壁对葡聚糖的通透性降低了10倍,并使TER增加了一倍。ADX后,即使在HS条件下,醛固酮也会降低隐窝壁葡聚糖通透性,增加TER,增加隐窝周围鞘中的钠积累以及ENaC表达。向ADX大鼠输注血管紧张素II并不能逆转醛固酮缺乏的影响。这些发现表明,仅醛固酮就导致了钠吸收的增加以及细胞旁和隐窝周围鞘通透性的降低。