thapsigargin and Hypertension--Renal

The negative functional effects of cyclic GMP are controlled by the sarcoplasmic reticulum calcium-ATPase (SERCA). The effects of cyclic GMP are blunted in cardiac hypertrophy. We tested the hypothesis that the interaction between cyclic GMP and SERCA would be reduced in hypertrophic cardiac myocytes. Myocytes were isolated from 7 control and 7 renal-hypertensive hypertrophic rabbits. Control and hypertrophic myocytes received 8-bromo-cGMP (8-Br-cGMP; 10(-7), 10(-6), 10(-5) mol/l), the SERCA blocker thapsigargin (10(-8) mol/l) followed by 8-Br-cGMP, or the SERCA blocker, cyclopiazonic acid (CPA; 10(-7) mol/l) followed by 8-Br-cGMP. Percent shortening and maximal rate of shortening and relaxation were recorded using a video edge detector. Changes in cytosolic Ca2+ were assessed in fura 2-loaded myocytes. In controls, 8-Br-cGMP caused a significant 36% decrease in percent shortening from 5.8 +/- 0.4 to 3.7 +/- 0.3%. Thapsigargin and CPA did not affect basal control or hypertrophic myocyte function. When 8-Br-cGMP was given following thapsigargin or CPA, the negative effects of 8-Br-cGMP on control myocyte function were reduced. In hypertrophic myocytes, 8-Br-cGMP caused a smaller but significant 17% decrease in percent shortening from 4.7 +/- 0.2 to 3.9 +/- 0.1%. When 8-Br-cGMP was given following thapsigargin or CPA, no significant changes occurred in hypertrophic cell function. Intracellular Ca2+ transients responded in a similar manner to changes in cell function in control and hypertrophic myocytes. These results show that the effects of cyclic GMP were reduced in hypertrophic myocytes, but this was not related to SERCA. In presence of SERCA inhibitors, the responses to cyclic GMP were blunted in hypertrophic as well as control myocytes.

Topics: Animals; Calcium; Cyclic GMP; Heart Ventricles; Hypertension, Renal; Hypertrophy; In Vitro Techniques; Indoles; Myocytes, Cardiac; Rabbits; Sarcoplasmic Reticulum Calcium-Transporting ATPases; Thapsigargin

This study examined whether alterations in intracellular or extracellular Ca2+ mobilization were related to differences in caffeine and phenylephrine (PHE)-induced contractions between two-kidney. one-clip hypertensive (2K-1C) and normotensive (2K) rat aortas. After depletion and reloading of intracellular Ca2+ stores, caffeine and PHE-induced contractions in Ca2+-free solution were increased in 2K-1C. Thapsigargin reduced the contraction to caffeine in 2K-1C and 2K with similar sensitivity. PHE-induced contraction in 1.6-mM Ca2+ solution was decreased in 2K-1C, and nifedipine was less effective in lowering this response. The responsiveness to extracellular Ca2+ was decreased in 2K-1C hypertensive rat aortas. Our results indicate an increased intracellular Ca2+ stores that are not related to alteration in Ca2+-ATPase function and a lower contribution of L-type channels to the contraction of 2K-1C aortas.

Topics: Animals; Aorta; Caffeine; Calcium; Calcium Channels, L-Type; Hypertension, Renal; In Vitro Techniques; Male; Nifedipine; Phenylephrine; Potassium Chloride; Rats; Rats, Wistar; Sarcoplasmic Reticulum; Thapsigargin; Vasoconstriction

1. The aim of the present study was to test in vitro if the increased contractile effect of phenylephrine and KCl, observed after the addition of ouabain in renal hypertensive rat aorta, is mediated by Ca2+ accumulated on the sarcoplasmic reticulum. 2. In aortas of one kidney (1K) rats, ouabain did not modify the concentration-effect curves stimulated with phenylephrine and KCl. 3. Contractile responses stimulated with phenylephrine and KCl were potentiated by ouabain in one kidney--one clip (1K-1C) aortas, and preincubation with thapsigargin abolished the increasing effect of ouabain on these contractions. 4. The addition of thapsigargin before phenylephrine and KCl did not modify the contractile response to phenylephrine and KCl or the resting vascular tone during the time incubation. 5. In the presence of ouabain, thapsigargin significantly increased the vascular tone only in 1K-1C rat aortas. 6. Increased intracellular Na+ concentration as a consequence of Na(+)-K(+)-ATPase inhibition induces increased accumulation of Ca2+ inside the sarcoplasmic reticulum in 1K-1C rat aortas. The differential effects in renal hypertensive and normotensive aortas suggest a possible role of this mechanism in modulating cytosolic Ca2+ in renal hypertension.

Topics: Animals; Aorta, Thoracic; Calcium-Transporting ATPases; Cardiotonic Agents; Enzyme Inhibitors; Hypertension, Renal; Male; Muscle Contraction; Muscle, Smooth, Vascular; Ouabain; Phenylephrine; Potassium Chloride; Rats; Rats, Wistar; Thapsigargin; Vasoconstrictor Agents