Effects of Roxadustat on Erythropoietin Production in the Rat Body

• Type of Material: Article
• Authors: Yukiko Yasuoka, Kitasato University; Yuichiro Izumi, Kumamoto University; Takashi Fukuyama, Kitasato University; Haruki Omiya, Iwate University; Truyen D Pham, Emory University; Hideki Inoue, Kumamoto University; Tomomi Oshima, Kitasato University; Taiga Yamazaki, Kitasato University; Takayuki Uematsu, Kitasato University; Noritada Kobayashi, Kitasato University; Yoshitaka Shimada, Kitasato University; Yasushi Nagaba, Kitasato University; Tetsuro Yamashita, Iwate University; Masashi Mukoyama, Kumamoto University; Yuichi Sato, Kitasato University; Susan Wall, Emory University; Jeff Sands, Emory University; Noriko Takahashi, Kitasato University; Katsumasa Kawahara, Kitasato University; Hiroshi Nonoguchi, Kitasato University
• Language: English
• Date: 2022-02-01
• Publisher: MDPI
• Publication Version: Final Published Version
• Copyright Statement: © 2022 by the authors.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.3390/molecules27031119
• Title of Journal or Parent Work: MOLECULES
• Volume: 27
• Issue: 3
• Grant/Funding Information: This study was supported by a Grant-in Aid for Scientific Research from the Ministry of Education, Culture, Sports, Sciences and Technology of Japan (26461259 (KK), 16K09654 (HN), 16K19493 (YI), 16K08505 (YY), 17K16578 (TF), 18K08247 (YI) and 19K09226 (TF)), the Science Research Promotion Fund from the Promotion, Kitasato University Medical Center: H25-003 (TF) and H24-011 (HN), Mutual Aid Corporation for Private Schools of Japan (HN) and by the NIH Grant to SMW (DK 119793).
• Keywords: Science & Technology; RENAL ANEMIA; INHIBITORS; Chemistry, Multidisciplinary; Chemistry; Western blotting; HIF2 alpha; proximal tubules; CELLS; hypoxia; Roxadustat; PHD inhibitor; EXPRESSION; Physical Sciences; GENE; erythropoietin; Life Sciences & Biomedicine; Biochemistry & Molecular Biology; HYPOXIA; deglycosylation; collecting ducts; RESPONSES
• Research Categories: Biology, Physiology; Health Sciences, Medicine and Surgery

Abstract

Anemia is a major complication of chronic renal failure. To treat this anemia, prolylhydrox-ylase domain enzyme (PHD) inhibitors as well as erythropoiesis-stimulating agents (ESAs) have been used. Although PHD inhibitors rapidly stimulate erythropoietin (Epo) production, the precise sites of Epo production following the administration of these drugs have not been identified. We developed a novel method for the detection of the Epo protein that employs deglycosylation-coupled Western blotting. With protein deglycosylation, tissue Epo contents can be quantified over an extremely wide range. Using this method, we examined the effects of the PHD inhibitor, Roxadustat (ROX), and severe hypoxia on Epo production in various tissues in rats. We observed that ROX increased Epo mRNA expression in both the kidneys and liver. However, Epo protein was detected in the kidneys but not in the liver. Epo protein was also detected in the salivary glands, spleen, epididymis and ovaries. However, both PHD inhibitors (ROX) and severe hypoxia increased the Epo protein abundance only in the kidneys. These data show that, while Epo is produced in many tissues, PHD inhibitors as well as severe hypoxia regulate Epo production only in the kidneys.