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α1-肾上腺素能受体亚型在心脏细胞培养中的表达

Expression of alpha1-adrenergic receptor subtypes in heart cell culture.

作者信息

Luther H P, Podlowski S, Schulze W, Morwinski R, Buchwalow I, Baumann G, Wallukat G

机构信息

Medical Clinic I, Department of Cardiology, Humboldt-University (Charité), Berlin, Germany.

出版信息

Mol Cell Biochem. 2001 Aug;224(1-2):69-79. doi: 10.1023/a:1011991117624.

Abstract

Three alpha1-AR subtypes have been cloned so far and are designated as alpha1a, alpha1b,, and alpha1d. Organ-specific distribution pattern and subtype-specific effects are known but not fully understood. To address a cell-type specific expression pattern in the heart we investigated expression pattern of alpha1-AR subtypes on RNA- and protein-level in heart tissue, cultured cardiomyocytes and non-myocytes of the rat. Each alpha1AR-subtype mRNA was present in neonatal and adult rat heart culture but the relative distribution pattern was significantly different. While the alpha1a-AR subtype is preferentially expressed in adult cardiomyocytes, the alpha1b-AR subtype was preferentially expressed in the non-myocyte cell fraction. The RT-PCR results were confirmed by Western-blotting (alpha1b) and immunocytochemical studies. Incubation with an alpha1-agonist (phenylephrine) for 72 h led to a significant reduction of the alpha1b-AR in neonatal heart cell culture on both mRNA and protein level. In contrast, incubation with an alpha1-antagonist (prazosin) induced a 1.6 fold upregulation of the alpha1a-AR mRNA without significant effects on radioligand binding and functional assay. The results indicate a distribution pattern of the alpha1-AR subtype which is specific for cell type and ontogeny of the rat heart and may be regulated by adrenergic agents.

摘要

迄今为止,已克隆出三种α1-肾上腺素能受体(AR)亚型,分别命名为α1a、α1b和α1d。器官特异性分布模式和亚型特异性效应已为人所知,但尚未完全了解。为了研究心脏中细胞类型特异性的表达模式,我们在RNA和蛋白质水平上研究了大鼠心脏组织、培养的心肌细胞和非心肌细胞中α1-AR亚型的表达模式。每种α1AR亚型的mRNA在新生大鼠和成年大鼠心脏培养物中均有表达,但相对分布模式存在显著差异。α1a-AR亚型在成年心肌细胞中优先表达,而α1b-AR亚型在非心肌细胞部分优先表达。RT-PCR结果通过蛋白质免疫印迹法(α1b)和免疫细胞化学研究得到证实。用α1-激动剂(去氧肾上腺素)孵育72小时导致新生心脏细胞培养物中α1b-AR在mRNA和蛋白质水平上显著降低。相反,用α1-拮抗剂(哌唑嗪)孵育诱导α1a-AR mRNA上调1.6倍,对放射性配体结合和功能测定无显著影响。结果表明,α1-AR亚型在大鼠心脏的细胞类型和个体发育中具有特异性分布模式,可能受肾上腺素能药物调节。

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