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A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement

Chronic hypoxia in the renal tubulointerstitium plays a key role in the progression of chronic kidney disease (CKD). It is therefore important to investigate tubular hypoxia and the activity of hypoxia‐inducible factor (HIF)‐1α in response to hypoxia. Rarefaction of the peritubular capillary causes...

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Autores principales: Honda, Tomoko, Hirakawa, Yosuke, Mizukami, Kiichi, Yoshihara, Toshitada, Tanaka, Tetsuhiro, Tobita, Seiji, Nangaku, Masaomi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7769172/
https://www.ncbi.nlm.nih.gov/pubmed/33369883
http://dx.doi.org/10.14814/phy2.14689
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author Honda, Tomoko
Hirakawa, Yosuke
Mizukami, Kiichi
Yoshihara, Toshitada
Tanaka, Tetsuhiro
Tobita, Seiji
Nangaku, Masaomi
author_facet Honda, Tomoko
Hirakawa, Yosuke
Mizukami, Kiichi
Yoshihara, Toshitada
Tanaka, Tetsuhiro
Tobita, Seiji
Nangaku, Masaomi
author_sort Honda, Tomoko
collection PubMed
description Chronic hypoxia in the renal tubulointerstitium plays a key role in the progression of chronic kidney disease (CKD). It is therefore important to investigate tubular hypoxia and the activity of hypoxia‐inducible factor (HIF)‐1α in response to hypoxia. Rarefaction of the peritubular capillary causes hypoperfusion in CKD; however, the effect of hypoperfusion on HIFs has rarely been investigated. We induced hypoperfusion caused by coverslip placement in human kidney‐2 cells, and observed an oxygen gradient under the coverslip. Immunocytochemistry of HIF‐1α showed a doughnut‐shaped formation on the edge of a pimonidazole‐positive area, which we named the “HIF‐ring”. The oxygen tension of the HIF‐ring was estimated to be between approximately 4 mmHg and 20 mmHg. This result was not compatible with those of past research showing HIF‐1α accumulation in the anoxic range with homogeneous oxygen tension. We further observed the presence of a pH gradient under a coverslip, as well as a shift of the HIF ring due to changes in the pH of the culture medium, suggesting that the HIF ring was formed by suppression of HIF‐1α related to low pH. This research demonstrated that HIF‐1α activation mimics the physiological state in cultured cells with hypoperfusion.
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spelling pubmed-77691722020-12-31 A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement Honda, Tomoko Hirakawa, Yosuke Mizukami, Kiichi Yoshihara, Toshitada Tanaka, Tetsuhiro Tobita, Seiji Nangaku, Masaomi Physiol Rep Original Research Chronic hypoxia in the renal tubulointerstitium plays a key role in the progression of chronic kidney disease (CKD). It is therefore important to investigate tubular hypoxia and the activity of hypoxia‐inducible factor (HIF)‐1α in response to hypoxia. Rarefaction of the peritubular capillary causes hypoperfusion in CKD; however, the effect of hypoperfusion on HIFs has rarely been investigated. We induced hypoperfusion caused by coverslip placement in human kidney‐2 cells, and observed an oxygen gradient under the coverslip. Immunocytochemistry of HIF‐1α showed a doughnut‐shaped formation on the edge of a pimonidazole‐positive area, which we named the “HIF‐ring”. The oxygen tension of the HIF‐ring was estimated to be between approximately 4 mmHg and 20 mmHg. This result was not compatible with those of past research showing HIF‐1α accumulation in the anoxic range with homogeneous oxygen tension. We further observed the presence of a pH gradient under a coverslip, as well as a shift of the HIF ring due to changes in the pH of the culture medium, suggesting that the HIF ring was formed by suppression of HIF‐1α related to low pH. This research demonstrated that HIF‐1α activation mimics the physiological state in cultured cells with hypoperfusion. John Wiley and Sons Inc. 2020-12-28 /pmc/articles/PMC7769172/ /pubmed/33369883 http://dx.doi.org/10.14814/phy2.14689 Text en © 2020 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research
Honda, Tomoko
Hirakawa, Yosuke
Mizukami, Kiichi
Yoshihara, Toshitada
Tanaka, Tetsuhiro
Tobita, Seiji
Nangaku, Masaomi
A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
title A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
title_full A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
title_fullStr A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
title_full_unstemmed A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
title_short A distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
title_sort distinctive distribution of hypoxia‐inducible factor‐1α in cultured renal tubular cells with hypoperfusion simulated by coverslip placement
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7769172/
https://www.ncbi.nlm.nih.gov/pubmed/33369883
http://dx.doi.org/10.14814/phy2.14689
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