PDF(Pubmed)
Abstract：
The intrarenal endothelin (ET) system is an established moderator of kidney physiology and mechanistic contributor to the pathophysiology and progression of chronic kidney disease in humans and rodents. The aim of the present study was to characterize ET system by combining single cell RNA sequencing (scRNA-seq) data with immunolocalization in human and rodent kidneys of both sexes. Using publicly available scRNA-seq data, we assessed sex and kidney disease status (human), age and sex (rats), and diurnal expression (mice) on the kidney ET system expression. In normal human biopsies of both sexes and in rodent kidney samples, the endothelin-converting enzyme-1 (ECE1) and ET-1 were prominent in the glomeruli and endothelium. These data agreed with the scRNA-seq data from these three species, with ECE1/Ece1 mRNA enriched in the endothelium. However, the EDN1/Edn1 gene (encodes ET-1) was rarely detected, even though it was immunolocalized within the kidneys, and plasma and urinary ET-1 excretion are easily measured. Within each species, there were some sex-specific differences. For example, in kidney biopsies from living donors, men had a greater glomerular endothelial cell endothelin receptor B (Ednrb) compared with women. In mice, females had greater kidney endothelial cell Ednrb than male mice. As commercially available antibodies did not work in all species, and RNA expression did not always correlate with protein levels, multiple approaches should be considered to maintain required rigor and reproducibility of the pre- and clinical studies evaluating the intrarenal ET system.
摘要：
肾内内皮素（ET）系统是肾脏生理学的既定调节剂，是人类和啮齿动物慢性肾脏疾病的病理生理学和进展的机制贡献者。这项研究的目的是通过将单细胞RNA测序（scRNA-seq）数据与两种性别的人类和啮齿动物肾脏中的免疫定位相结合来表征ET系统。使用公开可用的scRNA-seq数据，我们评估了性别和肾脏疾病状态（人类），年龄和性别（老鼠），和昼夜表达(小鼠)对肾脏ET系统的表达。在正常人的两性活检和啮齿动物的肾脏样本中，内皮素转换酶-1（ECE1）和ET-1在肾小球和内皮中突出。这些数据与来自这3个物种的scRNA-seq数据一致，ECE1/Ece1mRNA富集在内皮中。然而,EDN1/Edn1基因（编码ET-1）很少被检测到，即使它在肾脏中免疫定位，血浆和尿ET-1排泄量容易测量。在每个物种中，有一些性别差异。例如,在活体捐献者的肾脏活检中，男性的肾小球内皮细胞内皮素受体B(Ednrb)高于女性.在老鼠身上,雌性小鼠的肾内皮细胞Ednrb大于雄性小鼠。由于市售抗体并非在所有物种中均有效，RNA表达并不总是与蛋白质水平相关，应考虑多种方法来维持评估肾内ET系统的术前和临床研究所需的严谨性和可重复性.
