Effects of Roxadustat on Erythropoietin Production in the Rat Body

Yasuoka, Yukiko; Yokoyama, Izumi; Fukuyama, Takashi; Omiya, Haruki; Pham, Truyen D.; Inoue, Hideki; Oshima, Tomomi; Yamazaki, Taiga; Uematsu, Takayuki; Kobayashi, Noritada; Shimada, Yuji; Nagaba, Yasushi; Yamashita, Tetsuro; Mukoyama, Masashi; Sato, Yuichi; Wall, Susan M.; Sands, Jeff M.; Takahashi, Noriko; Kawahara, Katsumasa; Nonoguchi, Hiroshi

doi:10.3390/molecules27031119

Cited by 5 publications

(20 citation statements)

References 51 publications

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“… 10 By exposing rats to 7% oxygen, Yukiko Yasuoka et al found that roxadustat produced EPO in some tissues stimulated by hypoxia, specifically regulating EPO production in the kidney. 11 Hypoxia stimulates various adaptive responses, many of which are mediated through transcriptional complexes of the HIF family, directly upregulating the transcription of genes involved in erythropoiesis, angiogenesis, vasomotor tone, metabolic pathways, and processes related to cell reproduction and survival. What is the mechanism by which roxadustat counteracts high-altitude hypoxia?…”

Section: Discussionmentioning

confidence: 99%

A Pharmacodynamic Evaluation of the Protective Effects of Roxadustat Against Hypoxic Injury at High Altitude

Guo¹,

Li²,

Li³

et al. 2023

DDDT

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Purpose To investigate roxadustat’s preventive effects on hypoxia damage in the quick ascent to high altitude. Methods The roxadustat (7.8 mg/kg, 15.6 mg/kg, and 31.2 mg/kg) and control groups of BALB/C mice were distributed at random. To evaluate roxadustat’s anti-hypoxic effectiveness at the recommended dose, an atmospheric pressure closed hypoxic experiment was used. Wistar rats were randomly assigned to groups that received normal oxygen, hypoxic, acetazolamide, or roxadustat in order to evaluate the protective effects against hypoxic damage. Animal blood was obtained for arterial blood-gas analysis, inflammatory factors, and the identification of oxidative stress indicators. Animal tissues were removed for pathological investigation. Results In each group, the mice’s survival time was noticeably extended compared to the normal oxygen group. The medium dose had the best time extension rate at 19.05%. Blood SatO 2 and PaO 2 were significantly higher in the roxadustat group compared to the hypoxic group. Erythrocyte content, hemoglobin content, and hematocrit were also significantly higher. Plasma levels of IL-6, TNF-α, and IFN-γ were also significantly lower in the roxadustat group. Roxadustat can also improve the level of oxidative stress in the tissues of hypoxic rats. According to the results of HE staining, roxadustat could greatly lessen the harm done to rat heart, brain, lung, liver, and kidney tissue as a result of hypoxia. Conclusion Roxadustat can greatly reduce inflammation, oxidative stress, and tissue damage brought on by hypoxia, showing that it can significantly enhance the body’s ability to adapt to high altitude exposure.

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Section: Discussionmentioning

confidence: 99%

A Pharmacodynamic Evaluation of the Protective Effects of Roxadustat Against Hypoxic Injury at High Altitude

Guo¹,

Li²,

Li³

et al. 2023

DDDT

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“…This capillary Western blotting approach is also a useful method for the detection of Epo protein in both blood and urine. Instead of pre-purification, we introduced deglycosylation of the samples before Western blotting, as Desharnais P. et al did in combination with a capillary Western blotting [74,79,[86][87][88]. It is true that non-specific bands at around 25 and 50 kDa appear in our Western blotting, especially in blood, but those bands do not interfere with the appearance of glycosylated and deglycosylated Epo bands.…”

Section: Capillary Electromigration Methodsmentioning

confidence: 99%

“…Blood and/or urine samples are deglycosylated before our Western blotting [79,[86][87][88]]. An amount of 1 µL of 10% SDS is added to the 10 µL sample and boiled for 3 min at 100 Glycosylated and deglycosylated samples were applied to SDS-PAGE (10-20% gradient gel, 414893; Cosmo Bio, Tokyo, Japan).…”

Section: Methods Of Epo Detection By Deglycosylation-coupled Western ...mentioning

confidence: 99%

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Progress in the Detection of Erythropoietin in Blood, Urine, and Tissue

et al. 2023

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Detection of erythropoietin (Epo) was difficult until a method was developed by the World Anti-Doping Agency (WADA). WADA recommended the Western blot technique using isoelectric focusing (IEF)-PAGE to show that natural Epo and injected erythropoiesis-stimulating agents (ESAs) appear in different pH areas. Next, they used sodium N-lauroylsarcosinate (SAR)-PAGE for better differentiation of pegylated proteins, such as epoetin β pegol. Although WADA has recommended the use of pre-purification of samples, we developed a simple Western blotting method without pre-purification of samples. Instead of pre-purification, we used deglycosylation of samples before SDS-PAGE. The double detection of glycosylated and deglycosylated Epo bands increases the reliability of the detection of Epo protein. All of the endogenous Epo and exogenous ESAs shift to 22 kDa, except for Peg-bound epoetin β pegol. All endogenous Epo and exogenous ESAs were detected as 22 kDa deglycosylated Epo by liquid chromatography/mass spectrum (LC/MS) analysis. The most important factor for the detection of Epo is the selection of the antibody against Epo. WADA recommended clone AE7A5, and we used sc-9620. Both antibodies are useful for the detection of Epo protein by Western blotting.

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“…In addition, Karin M. Kirschner et al reported that the HIF-PHD like roxadustat can combat hypoxic partial pressure in the treatment of preterm infant diseases(Kirschner, Kelterborn et al 2022), such as bronchial dysplasia. Yukiko Yasuoka et al found that roxadustat produced EPO in a number of tissues stimulated by hypoxia by exposing rats to 7% oxygen, speci cally regulating EPO production in the kidney(Yasuoka, Izumi et al 2022).…”

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confidence: 99%

A Pharmacodynamic Evaluation of the Protective Effects of Roxadustat Against Hypoxic Injury at High Altitude

Guo

et al. 2022

Preprint

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Objectives: To explore the protective effects of roxadustat on hypoxia damage in the fast access to high altitude. Methods: BALB/C mice were randomly divided into control, roxadustat-7.8 mg/kg, roxadustat-15.6 mg/kg and roxadustat-31.2 mg/kg groups. The anti-hypoxic effectiveness of roxadustat in an optimal dose was evaluated by atmospheric pressure closed hypoxic experiment. Wistar rats were randomly divided into normal pressure, hypoxia model, acetazolamide and roxadustat groups to evaluate the protective effects against hypoxic damage. Animal blood was collected for arterial blood-gas analysis, cytokines detection, oxidative stress indicators, and their organs were harvested for pathological examination. Results: Compared with the control group, the survival time of mice were significantly prolonged in all groups. The time prolongation rate of the medium dose was 19.05%, which was the best dose. Compared with the hypoxia model group, the blood SatO2 and PaO2 in the roxadustat group were significantly increased; Erythrocyte content, hemoglobin content and hematocrit were significantly increased; Plasma levels of IL-6, TNF-α and IFN-γ were significantly decreased; MDA content in the myocardial, brain, lung and liver tissue were significantly decreased, SOD activity and GSH content in the tissue were significantly increased. The results of HE staining indicated that roxadustat could significantly improve the damage of heart, brain, lung, liver and kidney tissue after hypoxia in rats. Conlusions: Roxadustat can significantly prevent hypoxia-induced tissue damage, oxidative stress and inflammatory response indicating that roxadustat can obviously improve the adaptation to high-altitude exposure.

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Effects of Roxadustat on Erythropoietin Production in the Rat Body

Cited by 5 publications

References 51 publications

A Pharmacodynamic Evaluation of the Protective Effects of Roxadustat Against Hypoxic Injury at High Altitude

A Pharmacodynamic Evaluation of the Protective Effects of Roxadustat Against Hypoxic Injury at High Altitude

Progress in the Detection of Erythropoietin in Blood, Urine, and Tissue

A Pharmacodynamic Evaluation of the Protective Effects of Roxadustat Against Hypoxic Injury at High Altitude

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