Asberg A, Hartmann A, Attramadal H
Institute of Surgical Research, Medical Department B, The National Hospital, University of Oslo, Norway.
Scand J Clin Lab Invest. 1998 Jul;58(4):299-306. doi: 10.1080/00365519850186472.
The vasoconstrictive peptide endothelin-1 (ET-1) is an autocrine/paracrine peptide of putative pathophysiological importance in renal transplant medicine. The aim of the present study was to develop a method for analysis of gene expression of the renal endothelin system in humans. Only small amounts of tissue are available from renal cortical needle biopsies. Thus, in the present study we developed a quantitative assay based on the competitive reverse transcriptase polymerase chain reaction (RT-PCR) technology. We quantified endothelin A (ET(A)) and B (ET(B)) receptor subtype mRNAs and preproET-1 mRNA levels in renal cortex biopsies obtained before nephrectomy of healthy kidney donors. Mean (+/- SEM) mRNA levels of the ET(A) and ET(B) receptor subtypes in 26 living donors were 212 +/- 23 and 368 +/- 56 amol/microg total RNA, respectively. The preproET-1 mRNA level in 19 living donors was 213 +/- 28 amol/microg total RNA. The inter-assay coefficient of variation (CV) for the assay was 10%; the intra-assay CV was 6-13%. The competitive RT-PCR assay described provides an accurate tool for gene expression investigation of the human endothelin system in renal cortical needle biopsies.
血管收缩肽内皮素 -1（ET -1）是一种在肾移植医学中具有潜在病理生理重要性的自分泌/旁分泌肽。本研究的目的是开发一种分析人类肾内皮素系统基因表达的方法。肾皮质穿刺活检只能获取少量组织。因此，在本研究中，我们基于竞争性逆转录聚合酶链反应（RT -PCR）技术开发了一种定量检测方法。我们对健康肾供体肾切除术前获取的肾皮质活检组织中的内皮素A（ET(A)）和B（ET(B)）受体亚型mRNA以及前内皮素原 -1（preproET -1）mRNA水平进行了定量分析。26名活体供体中ET(A)和ET(B)受体亚型的平均（±SEM）mRNA水平分别为212±23和368±56 amol/μg总RNA。19名活体供体中的前内皮素原 -1 mRNA水平为213±28 amol/μg总RNA。该检测方法的批间变异系数（CV）为10%；批内CV为6 - 13%。所描述的竞争性RT -PCR检测方法为肾皮质穿刺活检中人类内皮素系统的基因表达研究提供了一种准确的工具。
