STIMULATION OF SODIUM-TRANSPORT BY ALDOSTERONE AND ARGININE VASOTOCIN IN A6 CELLS
STIMULATION OF SODIUM-TRANSPORT BY ALDOSTERONE AND ARGININE VASOTOCIN IN A6 CELLS
复制标题
DOI：
10.1016/0167-4889(88)90208-x
复制
发表时间：
1988-12-09
期刊：
影响因子：
--
通讯作者：
REIF, MC
中科院分区：
文献类型：
--
作者：
BINDELS, RJM;SCHAFER, JA;REIF, MC
本文观察了醛固酮和精氨酸缩血管素（AVT）对体外培养的A6细胞Na+跨上皮转运的影响。所有实验均使用在MillicellTM培养板插入物上生长2-4周的细胞在确定的无血清培养基中进行。在过滤器上接种细胞后2天省略胎牛血清不会影响电位差（PD）的发展或测试的激素反应。将细胞层置于Ussing室中用于短路电流（ISC）和跨上皮电导（G）测量。基线值为（n = 93）：PD，51.0 ±。0.2 mV（顶侧负）; ISC，14.55 ±。0.06μ A/cm 2; G，0306 ±。0.001 mS/cm 2。与对照相比，在培养箱中用10-7 M醛固酮预处理24小时的细胞中ISC和G更高（ISC，28 ± 0.01）。2对16 +-。2 μ A/cm 2，G，0.41 ±。0.04相对于0.26 ±。0.01 mS/cm 2，n = 5），并且两者均保持稳定至少6小时。在未用醛固酮处理的细胞中，10-7 M AVT在添加后1分钟内增加ISC，在15分钟内产生最大ISC，然后在接下来的5小时内下降到基线水平。向醛固酮预处理的细胞中加入AVT导致ISC比未预处理的细胞中显著更大的峰值增加（与对照相比ISC的变化：8.1 ± 0.01）。0.4 vs. 4.9 .+-。0.4μ A/cm 2，n = 5，P < 0.001），表明协同效应。在AVT存在下获得的阿米洛利的剂量-反应曲线显示阿米洛利完全抑制ISC。用醛甾酮预处理A6细胞24小时使阿米洛利剂量-反应曲线向右移动，表现为表观Ki值加倍（从0.17 ± 0.01）。0.02至0.33至0.04 μ M）。总之，A6细胞生长在确定的，无血清的培养基中表达的醛固酮和AVT之间的刺激跨上皮Na+转运大于加性协同作用。
The effects of aldosterone and arginine vasotocin (AVT) on transepithelial Na+ transport of cultured A6 cells were investigated. All experiments were performed with cells grown on MillicellTM culture-plate inserts for a period of 2-4 weeks in defined, serum-free medium. Omitting fetal bovine serum 2 days after seeding the cells on filters did not influence potential difference (PD) development or the hormonal responses tested. The cell layers were placed in an Ussing chamber for short-circuit current (ISC) and transepithelial conductance (G) measurements. Base-line values were (n = 93): PD, 51.0 .+-. 0.2 mV (apical side negative); ISC, 14.55 .+-. 0.06 .mu.A/cm2; G, 0306 .+-. 0.001 mS/cm2. ISC and G were higher in cells pretreated with 10-7 M aldosterone for 24 h in the incubator, when compared to controls (ISC, 28 .+-. 2 vs. 16 .+-. 2 .mu.A/cm2, G, 0.41 .+-. 0.04 vs. 0.26 .+-. 0.01 mS/cm2, n = 5) and both remained stable for at least 6 h. In cells not treated with aldosterone, 10-7 M AVT increased ISC within 1 min after addition, producing a maximum ISC within 15 min which then declined to baseline levels over the next 5 h. Addition of AVT to aldosterone-pretreated cells resulted in a significantly greater peak increase in ISC than in non-pretreated cells (change in ISC compared to controls: 8.1 .+-. 0.4 vs. 4.9 .+-. 0.4 .mu.A/cm2, n = 5, P < 0.001), indicating a synergistic effect. A dose-response curve for amiloride obtained in the presence of AVT showed that amiloride completely inhibits ISC. Pretreatment of the A6 cells with aldosteone for 24 h shifted the amiloride dose-response curve to the right, as expressed in a doubling of the apparent Ki value (from 0.17 .+-. 0.02 to 0.33 to 0.04 .mu.M). In conclusion, A6 cells grown in defined, serum-free medium express a greater than additive synergism between aldosterone and AVT in stimulating transepithelial Na+ transport.