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首页> 外文期刊>Canadian Journal of Physiology and Pharmacology >Endothelial cell-specific ETB receptor knockout: autoradiographic and histological characterisation and crucial role in the clearance of endothelin-1.
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Endothelial cell-specific ETB receptor knockout: autoradiographic and histological characterisation and crucial role in the clearance of endothelin-1.

机译:内皮细胞特异性ETB受体基因敲除:放射自显影和组织学特征以及在清除内皮素1中的关键作用。

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摘要

Inactivation of endothelin B receptors (ETB), either through selective pharmacological antagonism or genetic mutation, increases the circulating concentration of endothelin-1 (ET-1), suggesting ETB plays an important role in clearance of this peptide. However, the cellular site of ETB-mediated clearance has not yet been determined. We have used a novel mouse model of endothelial cell-specific knockout (KO) of ETB (EC ETB(-/-)) to evaluate the relative contribution of EC-ETB to the clearance of ET-1. Phenotypic evidence of EC-specific ETB KO was confirmed by immunocytochemistry and autoradiography. Binding of the radiolabelled selective ETB ligand BQ3020 was significantly and selectively decreased in EC-rich tissues of EC ETB(-/-) mice, including the lung, liver, and kidney. By contrast, ETA binding was unaltered. RT-PCR confirmed equal expression of ET-1 in tissue from EC ETB(-/-) mice and controls, despite increased concentration of plasma ET-1 in EC ETB(-/-). Clearance of an intravenous bolus of [(125)I]ET-1 was impaired in EC ETB(-/-) mice. Pretreatment with the selective ETB antagonist A192621 impaired [(125)I]ET-1 clearance in control animals to a similar extent, but did not further impair clearance in EC ETB(-/-) mice. These studies suggest that EC-ETB are largely responsible for the clearance of ET-1 from the circulation.
机译:通过选择性药理拮抗作用或基因突变使内皮素B受体(ETB)失活,会增加内皮素1(ET-1)的循环浓度,表明ETB在清除该肽中起着重要作用。然而,尚未确定ETB介导的清除的细胞部位。我们已经使用新型的ETB(EC ETB(-/-))内皮细胞特异性基因敲除(KO)小鼠模型来评估EC-ETB对清除ET-1的相对贡献。通过免疫细胞化学和放射自显影证实了EC特异性ETB KO的表型证据。放射性标记的选择性ETB配体BQ3020的结合在EC ETB(-/-)小鼠的富含EC的组织(包括肺,肝和肾)中显着且选择性降低。相反,ETA结合没有改变。尽管EC ETB(-/-)中血浆ET-1浓度增加,但RT-PCR证实了来自EC ETB(-/-)小鼠和对照的组织中ET-1的表达相等。 EC ETB(-/-)小鼠的[(125)I] ET-1静脉推注的清除受到损害。选择性ETB拮抗剂A192621的预处理在对照动物中损害[(125)I] ET-1的清除率达到相似的程度,但并未进一步损害EC ETB(-/-)小鼠的清除率。这些研究表明,EC-ETB很大程度上负责从循环中清除ET-1。

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