Effect of canrenone and amiloride on the prooxidative effect induced by aldosterone in human mononuclear leukocytes in vitro

2009 ◽  
Vol 32 (11) ◽  
pp. 895-898 ◽  
Author(s):  
C. Fiore ◽  
P. Sartorato ◽  
E. Pagnin ◽  
E. Ragazzi ◽  
L. A. Calò ◽  
...  
1992 ◽  
Vol 263 (5) ◽  
pp. E974-E979 ◽  
Author(s):  
M. Wehling ◽  
M. Christ ◽  
K. Theisen

Rapid nongenomic in vitro effects of aldosterone on intracellular electrolytes, cell volume, and Na(+)-H+ antiport have been found in human mononuclear leukocytes (HML). Binding of 125I-labeled aldosterone to plasma membranes of HML shares important features with these functional data. This includes a very low apparent dissociation constant (Kd) of 0.1 nM for both aldosterone and the effect on the Na(+)-H(+)-antiport, a high turnover rate, and the almost exclusive binding selectivity for aldosterone. Dexamethasone, RU 26988, corticosterone, ouabain, amiloride, and 18-hydroxyprogesterone were inactive as ligands. Deoxycorticosterone acetate had an intermediate activity with an apparent Kd of 100 nM. These findings are the first to demonstrate membrane binding of aldosterone being compatible with major aspects of its nongenomic effects.


Blood ◽  
1979 ◽  
Vol 54 (2) ◽  
pp. 359-370 ◽  
Author(s):  
RL Edwards ◽  
FR Rickles ◽  
AM Bobrove

Abstract Human mononuclear leukocytes generate the procoagulant material tissue factor (TF) following stimulation by endotoxin, mitogens, or antigens in vitro. We have examined tissue-factor generation by mononuclear cell subpopulations prepared in a variety of ways in order to determine the cell of origin of mononuclear cell TF and the conditions necessary for maximal in vitro TF generation. We have also examined the relationship between in vitro TF generation and in vivo or in vitro measures of delayed hypersensitivity in response to identical antigen stimulation. Our results demonstrate that the monocyte is responsible for the bulk of mononuclear cell TF generation in vitro and that adhesion alone is not sufficient stimulation for significant.


1987 ◽  
Vol 252 (4) ◽  
pp. E505-E508 ◽  
Author(s):  
M. Wehling ◽  
D. Armanini ◽  
T. Strasser ◽  
P. C. Weber

The in vitro effect of aldosterone on intracellular sodium and potassium concentration was investigated in human mononuclear leukocytes (HML). HML were separated from blood by a Percoll gradient and intracellular Na+ and K+ were determined before and after incubation for 1 h at 37 degrees C. The intracellular (ic) Na+ and K+ concentrations after separation of HML were 17 +/- 5 and 59 +/- 18 mmol/kg wet cells (mean +/- SD, n = 6), respectively. In the absence of aldosterone the ic Na+ concentration decreased to 12 +/- 4, whereas the ic Na+ concentration remained constant at 18 +/- 8 mmol/kg wet cells when aldosterone (1.4 nM) was added to the incubation medium. In parallel, the ic K+ concentration decreased without added aldosterone but remained constant with aldosterone. The effect of aldosterone on the ic Na+ and K+ concentrations of HML was blocked by the aldosterone antagonist canrenone (140 nM). Cortisol at a physiological concentration (40 nM) did not alter ic Na and K concentrations in these cells. The results suggest that aldosterone binding to specific receptors in human mononuclear leukocytes significantly contributes to the regulation of monovalent cation levels in these cells. Thus, this easily accessible model of HML allows study of the effects of mineralocorticoids in relation to their receptors in intact cells in normal and pathological states.


1978 ◽  
Vol 147 (2) ◽  
pp. 324-339 ◽  
Author(s):  
D G Nathan ◽  
L Chess ◽  
D G Hillman ◽  
B Clarke ◽  
J Breard ◽  
...  

Human mononuclear leukocytes were fractionated into populations of null, T and B cells by immunoabsorbent column chromatography followed by E-rosette formation and purification of T cells by differential centrifugation and osmotic lysis. The unfractionated and fractionated cell populations were first separately cultured for 14 days in plasma clots in the presence of two international units erythropoietin. Typical erythroid burst-forming unit (BFU-E)-derived colonies grew in the unfractionated cell cultures but not from T- or B-cell cultures. BFU-E colonies grew in null cell cultures but most of the colonies were small and variably hemoglobinized with less than three subcolonies. When intact T cells were added to null cells and cocultured, many typical large BFU-E colonies with more than 10 well homogenized subcolonies appeared. Increasing numbers of large BFU-E colonies in null cell cultures were induced by stepwise addition of T cells but not by the addition of B cells. A conditioned medium in which T cells had been induced to divide by tetanus toxoid substituted for intact T cells in this T-cell-dependent BFU-E colony formation observed in null cells. These findings demonstrate that the BFU-E, a committeded erythroid stem cell, resides in the null cell fraction of peripheral blood, but its proliferative capacity and differentiation in vitro requires a soluble product of T cells. Such experiments now permit a new approach to the assessment of various disorders of erythropoiesis. Erythroid hypoplasia in a particular case may be due to dysfunction of the committed precursor cell or to a failure of a helper effect induced by T cells.


1989 ◽  
Vol 257 (2) ◽  
pp. E170-E174 ◽  
Author(s):  
M. Wehling ◽  
S. Kuhls ◽  
D. Armanini

In vitro binding of aldosterone to mineralocorticoid receptors on human mononuclear leukocytes (HML) and its effects on the intracellular sodium and potassium concentrations of HML have already been described. In the present paper this easily accessible human cell model was investigated with regard to the regulation of the cell volume by aldosterone, since the concordant changes of sodium and potassium were expected to be accompanied by water and volume shifts. As determined by the measurement of cell diameter and the planimetric estimation of cell area in photographs, cell volume decreased by approximately 16% when cells were incubated in RPMI-1640 medium without aldosterone added for 1 h at 37 degrees C, a decrease not seen when 1.4 nM aldosterone was added to the incubation medium; the effect was half maximal at a concentration between 0.07 and 0.14 nM. One hundred forty nanomoles canrenone antagonized the action of aldosterone, but cortisol was ineffective. The results indicate concordant changes of intracellular sodium and potassium and cell volume, if studied under the same conditions. These data are the first to demonstrate that aldosterone is a major physiological determinant of lymphocyte volume in isotonic media.


Blood ◽  
1979 ◽  
Vol 54 (2) ◽  
pp. 359-370 ◽  
Author(s):  
RL Edwards ◽  
FR Rickles ◽  
AM Bobrove

Human mononuclear leukocytes generate the procoagulant material tissue factor (TF) following stimulation by endotoxin, mitogens, or antigens in vitro. We have examined tissue-factor generation by mononuclear cell subpopulations prepared in a variety of ways in order to determine the cell of origin of mononuclear cell TF and the conditions necessary for maximal in vitro TF generation. We have also examined the relationship between in vitro TF generation and in vivo or in vitro measures of delayed hypersensitivity in response to identical antigen stimulation. Our results demonstrate that the monocyte is responsible for the bulk of mononuclear cell TF generation in vitro and that adhesion alone is not sufficient stimulation for significant.


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