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The effects of nonselective ETA/ETB receptor blockade with intravenous bolus injection of bosentan (10 mg/kg) on renal excretory function and blood pressure were investigated in conscious, male, normotensive Wistar rats before and one week after bilateral renal denervation. Renal denervation was followed by an increase in urine flow rate from 4.54±0.38 to 5.72±0.36 µl/min.100 g b.w. (p<0.05) and a decrease in urine osmolality from 855.5±44.6 to 707.4±47.5 mosm/kg H2O (p<0.05). Bosentan administration in sham-operated rats resulted in decrease in urine flow rate from 4.54±0.38 to 3.49±0.34 µl/min.100 g b.w. (p<0.05), and increase in urine osmolality from 855.5±44.6 to 1075.0±76.1 mosm/kg H2O (p<0.05). Sodium excretion decreased from 226.9±20.0 to 155.1±11.0 nmol/min.100 g b.w. (p<0.01). Bosentan administration in renal denervated rats did not produce any changes in renal water or electrolyte excretions. Blood pressure, heart rate, clearance of Inulin or clearance of paraaminohippuric acid (PAH) did not change in sham-operated or renal denervated rats during nonselective ETA/ETB receptor blockade. Bosentan did not alter the baroreflex sensitivity or sympatho-vagal balance in sham-operated or renal denervated rats. In conclusion, an interaction between renal nerves and endothelins appears to be involved in the regulation of the renal excretory function.
Endothelin (ET), originally characterized as a vasoconstrictive peptide, has been found to have many different biological functions, including acting as a local hormonal regulator of pressure, fluid, ions and neurotransmitters in the inner ear. The objective of this study was to examine and quantify the mRNA expression of the endothelin type A and B receptors (ETAR and ETBR) in the strial vascularies (StV) and non-strial tissues (NSt) of the cochlear lateral wall using the real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique. The mouse tissue samples were harvested and RNA was extracted. RT was performed to obtain cDNA, and then the mRNA expression of each gene was measured via real-time PCR. We found that both receptor subtypes were expressed in the cochlear lateral wall, with a predominance of ETAR over ETBR. We showed that the mRNA expression of the two receptor subtypes was higher in the StV with a 1.8 times higher level of ETAR and an 8.1 times higher level of ETBR mRNAs than in the adjacent NSt of the lateral wall tissue. This study shows the existence and the quantity of ET receptor subtypes in the StV and NSt of the mouse cochlea. Our results suggest that an endothelin-mediated response via two different receptors, ETAR and ETBR, may play an important role in the physiological functions of the cochlear lateral wall by maintaining the homeostatic environment of the cochlea.
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Pathways of glomerular toxicity of cyclosporine-A: an "in vitro" study

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Background/Aims. Knowledge of renal toxicity of cyclosporine-A (CyA) is clouded by multiple effects on different glomerular and tubular cells and on kidney and systemic hemodynamics. To focus on glomerular action of CyA we used glomeruli isolated in vitro, with the aim of dissecting the effects on recruitment of glomerular vasoconstricting systems, like endothelin-1 (ET) and angiotensins (AI and AII). Methods. We studied the pathways of CyA damage on pig glomeruli isolated in vitro with the technique of sieving through mesh filters of different sizes, and incubated in an appropriate culture medium. The supernatant was sampled at different time intervals to measure ET, AI and AII concentrations upon addition of ET 10-12 or CyA 4·10-7 M, with or without either selective endothelin receptor A (ETA) or B (ETB), or unselective ETA-ETB receptor inhibitors. Results. CyA increased ET concentration (from 9.7±0.3 to 11.4±0.4 pg·ml-1, p<0.002), and the added ET released AI in the medium (from 26.6±4.7 to 39.1±4.6 pg·ml-1, p<0.05) when ETB receptors were blocked. In contrast, CyA stimulated angiotensins release independent of ET receptors blockade, hence, irrespective of ET concentration in the medium, from 26.6±4.7 to 38.0±2.1 pg·ml-1 for AI, p<0.05, and from 12.3±1.0 to 14.8±0.9 pg·ml-1 for AII, p<0.05. Conclusion. CyA releases ET and angiotensins independently by a direct action. Glomerular CyA toxicity might be mediated by recruitment of vasoconstricting peptides and modulated by relative ETA and ETB receptor occupancy.
The role of renal nerves in the effects of concomitant NO synthase and non-selective ETA/ETB receptor inhibition on renal function was investigated in conscious normotensive Wistar rats. NO synthase inhibition alone (10 mg/kg b. w. i.v. L-NAME) in sham-operated rats with intact renal nerves induced an increase in systolic, diastolic and mean arterial pressure, urine flow rate, sodium, chloride and calcium excretion (p<0.05). The effect of L-NAME was markedly reduced by bosentan (10 mg/kg b.w. i.v.) and the values of urine flow rate, sodium, chloride and calcium excretions returned to control level (p<0.05). L-NAME administration one week after a bilateral renal denervation increased blood pressure to a similar extent as in sham-operated rats but decreased urine flow rate (p<0.05) and did not change electrolyte excretion. ETA/ETB receptor inhibition with bosentan during NO synthase inhibition in the renal denervated rats did not produce changes in urine flow rate or electrolyte excretion. NO synthase inhibition as well as concurrent NO synthase and ETA/ETB receptor inhibition did not change clearance of inulin or paraaminohippuric acid in sham-operated or renal denervated rats. These results indicate that renal sympathetic nerves play an important modulatory role in NO and endothelin induced effects on renal excretory function.
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