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. 2018 Mar 13;8:4448. doi: 10.1038/s41598-018-22791-y

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Association of tubular reabsorption with EPO in neonatal kidneys. (a–e) The expression of tubular transporters at P7 was lower than that at P14 (n = 6). (f–k) Inhibition of reabsorption results in reduced renal EPO production. (f) The expression of EPO mRNA decreased 90 minutes after the use of diuretics (combination of acetazolamide, furosemide, hydrochlorothiazide, and spironolactone) (n = 4). (g–k) Use of losartan, which reduces glomerular filtration and tubular reabsorption, elevated renal PtO₂ (g), and reduced pimonidazole-positive hypoxic area (h and i), EPO levels (j), and RBC counts, hematocrit levels, and hemoglobin levels (k) (n = 4). (l–t) Enhancing reabsorption results in increased EPO production. Long-term use of diuretics (combination of acetazolamide and furosemide) with normal saline supplement from P3 lowered renal PtO₂ (l), and increased pimonidazole-positive hypoxic area (m and n), EPO levels (o), RBC counts, hematocrit levels, and hemoglobin levels (p) at P7 compared with the control. The expression of sodium transporters was elevated in the diuretics group (q–t) (n = 4). Error bars indicate SEM. *P < 0.05 by t-test or Tukey-Kramer test. n.s., not significant. Scale bars denote 100 micrometer. In the graphs of mRNA expression, arbitrary unit is used for vertical axis. RBC, red blood cells; D, diuretics; Ctrl, control; Los, losartan.