Expression and Function of Endothelins, Endothelin Receptors, and  Endothelin Converting Enzyme in the Porcine Trachea

Yoshimura, Hayashi; Nishimura, Junji; Sakihara, Chie; Kobayashi, Sei; Takahashi, Shosuke; Kobayashi, Hideo

doi:10.1165/ajrcmb.17.4.2832

Cited by 13 publications

(6 citation statements)

References 48 publications

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“…In the current study, virus‐induced increases in ir‐ET levels in the lung were markedly attenuated by CGS 26303, an ECE inhibitor that has recently been shown to inhibit ET‐1 synthesis in cultured guinea‐pig tracheal epithelial cells [14]. The combination of these data indicate that virus‐associated increases in ir‐ET content were due, at least in part, to enhanced synthesis of ET by the airway epithelium, although recent studies indicate that tracheal smooth muscle cells can also express prepro‐ET‐1 and ECE‐1 messenger ribonucleic acid (mRNA) [15]. Although the underlying mechanism of virus‐induced increases in ET content is unknown, an increased expression of ET‐1 mRNA is likely to be involved [6].…”

Section: Discussionmentioning

confidence: 99%

The role of endothelin in mediating virus‐induced changes in endothelinB receptor density in mouse airways

Henry¹,

Carr²,

Goldie³

et al. 1999

Eur Respir J

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Emerging evidence supports a mediator role for endothelin (ET)-1 in airway diseases including asthma. Respiratory tract viral infections, are associated with increased levels of ET and altered ET receptor density and function in murine airways. To determine whether these virus-induced effects are causally linked, perhaps involving ET-1-induced ET B receptor downregulation, the current study investigated the influence of in vivo administration of CGS 26303, an ET-converting enzyme inhibitor, on virus-induced changes in ET-content and ET B receptor density.CGS 26303 (5 mg . kg -1 . day -1) or placebo was administered to mice via osmotic minipumps implanted subcutaneously. Two days after implantation, mice were inoculated with influenza A/PR-8/34 virus or sham-infected, and all measurements were performed on tissue obtained on the fourth day post-inoculation.Viral infection was associated with elevated levels of immunoreactive ET and decreased densities of ET B receptors in murine airways. Both of these effects were attenuated in virus-infected mice that had received CGS 26303. Virus-induced increases in wet lung weight were also inhibited by CGS 26303. Importantly, administration of CGS 26303 had no effect on the titres of infectious virus in the lungs and similarly, viral infection had no effect on the plasma levels of free CGS 26303.In summary, CGS 26303 inhibited the virus-induced changes in both immunoreactive endothelin content and endothelin B receptor density. These findings are consistent with the postulate that the elevated epithelial expression of endothelin-1 during respiratory tract viral infection is a contributing factor in the downregulation of endothelin B receptors in airway smooth muscle. Whether inhibitors of endothelin synthesis attenuate virus-induced exacerbations of asthma or airways hyperresponsiveness remains to be established. Eur Respir J 1999; 14: 92±97. The levels of endothelin (ET)-1 in the airways are significantly elevated in respiratory diseases such as asthma [1] as well as in several animal models of airways disease, including allergic inflammation [2,3] and respiratory tract viral infection [4]. ET-1, via its potent actions on a raft of different cell types within the airways and lungs (for review see [5]), may contribute significantly to airway wall remodelling, oedema, bronchial obstruction, long-lasting bronchoconstriction and the development of airway hyperresponsiveness in asthma and during respiratory tract viral infections.Respiratory syncitial virus induces the expression of ET-1 in bronchial epithelial cells [6] and the levels of immunoreactive (ir)-ET-1 are elevated in murine airways and lung during influenza A viral infection [4]. Increases in the production of ET-1 by the airway epithelium would be expected to lead to enhanced stimulation of ET A and ET B receptors in adjacent tissues such as the airway smooth muscle. However, respiratory tract viral infection in mice was associated with reductions in ET B receptor density, which was reflected in attenuated...

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Section: Discussionmentioning

confidence: 99%

The role of endothelin in mediating virus‐induced changes in endothelinB receptor density in mouse airways

Henry¹,

Carr²,

Goldie³

et al. 1999

Eur Respir J

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“…The PCR amplification of preproET‐1 cDNA was performed in a 25‐ μ L reaction volume with 0.25 unit of Taq polymerase (Roche Diagnostics) and corresponding PCR buffer (final concentrations: 10 m M Tris–HCl, 50 m M KCl, pH 8.3), 2.5 m M MgCl 2 , 0.2 m M of each dNTP and 15 pmol of each primer (sequences as published: Yoshimura et al . (1997), using 1 μ L of reverse transcription product as template.…”

Section: Methodsmentioning

confidence: 99%

Endothelin synthesis and receptors in porcine kidney

Bäcker

Bokemeyer

Kramer

2001

Acta Physiologica Scandinavica

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As insufficient information on the endothelin (ET) system in the porcine kidney is available at present, we investigated renal ET-1 synthesis and ET receptors in this species. Because ET specifically affects renal and glomerular haemodynamics and distal tubular reabsorption, we studied ET-1 synthesis in isolated glomeruli and in inner medullary collecting duct (IMCD) cells and preproET-1 mRNA in renal cortex, isolated glomeruli and papillary tissue. In addition, we characterized density and properties of ET receptors in membranes from isolated glomeruli and papillary tissue. In contrast to isolated IMCD cells, which synthesized 120 +/- 11 fmol h(-1) mg-1 protein of ET-1, no such synthesis was found with isolated glomeruli in our assay system. Nevertheless, with RT-PCR preproET(-1) mRNA was clearly present in renal cortex and glomeruli as well as in papillary tissue. Glomerular membranes were found to have ET receptors with Bmax of 1.6 +/- 0.2 pmol mg-1 protein and Kd of 311 +/- 33 pmol L(-1). Using BQ-123 (10-5 M), a specific blocker of ETA receptors, we found that 58% of total receptors are ETA receptors. Thus, presumably 42% are ETB receptors (Bmax 0.7 +/- 0.1 pmol mg-1 protein; Kd 429 +/- 110 pmol L(-1)). Bosentan (10-5 M), an ETA- and ETB-receptor antagonist, blocked all ET receptors in glomerular membranes. Papillary membranes showed ET receptors with Bmax of 2.1 +/- 0.2 pmol mg-1 protein and Kd of 137 +/- 11 pmol L(-1). In the presence of BQ-123 (10-5 M) we found that all receptors are ETB receptors (Bmax 2.3 +/- 0.4 pmol mg-1 protein; Kd 162 +/- 25 pmol L(-1)). Bosentan (10-5 M) again blocked all ET receptors in papillary membranes, thus confirming our previous finding that IMCD cells possess high-affinity ETB receptors mediating the diuretic effects of ET. Thus, in the porcine kidney the ET system may act in an autocrine/paracrine manner at the glomerular as well as at the IMCD level.

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“…Primers for ET-1 and the report gene, HPRT, were already used in our laboratory. Except for endothelin converting enzyme (ECE)-1 and inducible NO synthase (iNOS), 14,15 the previously reported end-point PCR primers were unadapted to the Sybr Green RTQ-PCR system. To develop specific porcine primers adapted to Sybr Green RTQ-PCR conditions, we engaged classic primers for ET A , ET B , neuronal NO synthase (nNOS), endothelial NO synthase (eNOS), and VEGF in end-point PCR (GeneAmp PCR system 2400) to amplify cDNA from pig tissue.…”

Section: Real-time Quantification Pcrmentioning

confidence: 99%

Bosentan for the Prevention of Overcirculation-Induced Experimental Pulmonary Arterial Hypertension

et al. 2003

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Background-The dual endothelin-receptor antagonist bosentan has been reported to improve pulmonary arterial hypertension, but the role of endothelins in the pathogenesis of the condition remains uncertain. We investigated the roles of endothelin-1 (ET-1), nitric oxide (NO), vascular endothelial growth factor (VEGF), and tenascin in overcirculation-induced pulmonary hypertension in piglets, as a model of early pulmonary arterial hypertension, with or without bosentan therapy. Methods and Results-Thirty 3-week-old piglets were randomized to placebo or to bosentan 15 mg/kg BID after the anastomosis of the left subclavian artery to the pulmonary arterial trunk or after a sham operation. Three months later, the animals underwent a hemodynamic evaluation followed by cardiac and pulmonary tissue sampling for morphometry, immunohistochemistry, and real-time quantitative PCR. Chronic systemic-to-pulmonary shunting increased circulating plasma ET-1, pulmonary mRNA for ET-1, ET B receptor, inducible NO synthase, VEGF, and pulmonary ET-1 and VEGF proteins. There were increases in myocardial mRNA for ET A receptor and VEGF and in myocardial VEGF protein. Pulmonary and myocardial tissue mRNA for tenascin did not change. Normalized-flow pulmonary artery pressure increased from 20 (2) to 33 (1) mm Hg [mean (SEM)], arteriolar medial thickness increased on average by 83%, and these changes were completely prevented by bosentan therapy. Right ventricular end-systolic elastance increased in proportion to pulmonary arterial elastance with or without bosentan. Conclusions-Experimental overcirculation-induced pulmonary arterial hypertension appears to be causally related to an activation of the pulmonary ET-1 system and as such is completely prevented by the dual endothelin receptor antagonist bosentan. (Circulation. 2003;107:1329-1335.)

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Expression and Function of Endothelins, Endothelin Receptors, and Endothelin Converting Enzyme in the Porcine Trachea

Cited by 13 publications

References 48 publications

The role of endothelin in mediating virus‐induced changes in endothelinB receptor density in mouse airways

The role of endothelin in mediating virus‐induced changes in endothelinB receptor density in mouse airways

Endothelin synthesis and receptors in porcine kidney

Bosentan for the Prevention of Overcirculation-Induced Experimental Pulmonary Arterial Hypertension

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