Development of enzyme-linked immunosorbent assays for urinary thiazide-sensitive Na-Cl cotransporter measurement

Kiyoshi Isobe, Takayasu Mori, Takako Asano, Hiroyuki Kawaguchi, Shigeaki Nonoyama, Naonori Kumagai, Fumiaki Kamada, Tetsuji Morimoto, Matsuhiko Hayashi, Eisei Sohara, Tatemitsu Rai, Sei Sasaki, Shinichi Uchida

研究成果: Article査読

26 被引用数 (Scopus)

抄録

The Na-Cl cotransporter (NCC) in the distal convoluted tubules in kidney is known to be excreted in urine. However, its clinical significance has not been established because of the lack of quantitative data on urinary NCC. We developed highly sensitive enzymelinked immunosorbent assays (ELISAs) for urinary total NCC (tNCC) and its active form, phosphorylated NCC (pNCC). We first measured the excretion of tNCC and pT55-NCC in urinary exosomes in pseudohypoaldosteronism type II (PHAII) patients since PHAII is caused by NCC activation. Highly increased excretion of tNCC and pNCC was observed in PHAII patients. In contrast, the levels of tNCC and pNCC in the urine of patients with Gitelman's syndrome were not detectable or very low, indicating that both assays could specifically detect the changes in urinary NCC excretion caused by the changes of NCC activity in the kidney. Then, to test whether these assays could be feasible for a more general patient population, we measured tNCC and pNCC in the urine of outpatients with different clinical backgrounds. Although urinary protein levels >30 mg/dl interfered with our ELISA, we could measure urinary pNCC in all patients without proteinuria. Thus we established highly sensitive and quantitative assays for urinary NCC, which could be valuable tools for estimating NCC activity in vivo.

本文言語English
ページ(範囲)F1374-F1381
ジャーナルAmerican Journal of Physiology - Renal Physiology
305
9
DOI
出版ステータスPublished - 01-11-2013
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生理学
  • 泌尿器学

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