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Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells

Background: A low level of serum magnesium ion (Mg(2+)) is associated with type 2 diabetes mellitus (T2D). However, the molecular mechanism of Mg(2+) deficiency has not been fully clarified. The current study sought to assesses the effect of reactive oxygen species on the expression of Mg(2+) channe...

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Autores principales: Hirota, Chieko, Takashina, Yui, Yoshino, Yuta, Hasegawa, Hajime, Okamoto, Ema, Matsunaga, Toshiyuki, Ikari, Akira
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8393788/
https://www.ncbi.nlm.nih.gov/pubmed/34440664
http://dx.doi.org/10.3390/cells10081893
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author Hirota, Chieko
Takashina, Yui
Yoshino, Yuta
Hasegawa, Hajime
Okamoto, Ema
Matsunaga, Toshiyuki
Ikari, Akira
author_facet Hirota, Chieko
Takashina, Yui
Yoshino, Yuta
Hasegawa, Hajime
Okamoto, Ema
Matsunaga, Toshiyuki
Ikari, Akira
author_sort Hirota, Chieko
collection PubMed
description Background: A low level of serum magnesium ion (Mg(2+)) is associated with type 2 diabetes mellitus (T2D). However, the molecular mechanism of Mg(2+) deficiency has not been fully clarified. The current study sought to assesses the effect of reactive oxygen species on the expression of Mg(2+) channels and miRNA. Methods: The expression of Mg(2+) channels and miRNA were examined by real-time polymerase chain reaction. Intracellular Mg(2+) concentration was measured by Magnesium Green fluorescence measurement. Results: The mRNA level of transient receptor potential melastatin 6 (TRPM6), which functions as Mg(2+) influx channel in the distal convoluted tubule (DCT) of the kidney, was decreased by glycated albumin (GA), but not by insulin in rat renal tubule-derived NRK-52E cells. The mRNA levels of TRPM7, a homologue of TRPM6, and CNNM2, a Mg(2+) efflux transporter located at the basolateral membrane of DCT, were changed by neither GA nor insulin. The generation of reactive oxygen species (ROS) was increased by GA. Hydrogen peroxide (H(2)O(2)) dose-dependently decreased TRPM6 mRNA, but it inversely increased the reporter activity of TRPM6. H(2)O(2) accelerated the degradation of TRPM6 mRNA in actinomycin D assay without affecting TRPM7 and CNNM2 mRNA expressions. Nine miRNAs were considered as candidates for the regulator of stability of TRPM6 mRNA. Among them, miR-24-3p expression was increased by H(2)O(2). The H(2)O(2)-induced reduction of TRPM6 mRNA was rescued by miR-24-3p siRNA. Magnesium Green fluorescence measurement showed that Mg(2+) influx is suppressed by H(2)O(2), which was rescued by an antioxidant and miR-24-3p siRNA. Conclusions: We suggest that GA decreases TRPM6 expression mediated by the elevation of ROS and miR-24-3p in renal tubular epithelial cells of T2D.
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spelling pubmed-83937882021-08-28 Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells Hirota, Chieko Takashina, Yui Yoshino, Yuta Hasegawa, Hajime Okamoto, Ema Matsunaga, Toshiyuki Ikari, Akira Cells Article Background: A low level of serum magnesium ion (Mg(2+)) is associated with type 2 diabetes mellitus (T2D). However, the molecular mechanism of Mg(2+) deficiency has not been fully clarified. The current study sought to assesses the effect of reactive oxygen species on the expression of Mg(2+) channels and miRNA. Methods: The expression of Mg(2+) channels and miRNA were examined by real-time polymerase chain reaction. Intracellular Mg(2+) concentration was measured by Magnesium Green fluorescence measurement. Results: The mRNA level of transient receptor potential melastatin 6 (TRPM6), which functions as Mg(2+) influx channel in the distal convoluted tubule (DCT) of the kidney, was decreased by glycated albumin (GA), but not by insulin in rat renal tubule-derived NRK-52E cells. The mRNA levels of TRPM7, a homologue of TRPM6, and CNNM2, a Mg(2+) efflux transporter located at the basolateral membrane of DCT, were changed by neither GA nor insulin. The generation of reactive oxygen species (ROS) was increased by GA. Hydrogen peroxide (H(2)O(2)) dose-dependently decreased TRPM6 mRNA, but it inversely increased the reporter activity of TRPM6. H(2)O(2) accelerated the degradation of TRPM6 mRNA in actinomycin D assay without affecting TRPM7 and CNNM2 mRNA expressions. Nine miRNAs were considered as candidates for the regulator of stability of TRPM6 mRNA. Among them, miR-24-3p expression was increased by H(2)O(2). The H(2)O(2)-induced reduction of TRPM6 mRNA was rescued by miR-24-3p siRNA. Magnesium Green fluorescence measurement showed that Mg(2+) influx is suppressed by H(2)O(2), which was rescued by an antioxidant and miR-24-3p siRNA. Conclusions: We suggest that GA decreases TRPM6 expression mediated by the elevation of ROS and miR-24-3p in renal tubular epithelial cells of T2D. MDPI 2021-07-26 /pmc/articles/PMC8393788/ /pubmed/34440664 http://dx.doi.org/10.3390/cells10081893 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hirota, Chieko
Takashina, Yui
Yoshino, Yuta
Hasegawa, Hajime
Okamoto, Ema
Matsunaga, Toshiyuki
Ikari, Akira
Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells
title Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells
title_full Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells
title_fullStr Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells
title_full_unstemmed Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells
title_short Reactive Oxygen Species Downregulate Transient Receptor Potential Melastatin 6 Expression Mediated by the Elevation of miR-24-3p in Renal Tubular Epithelial Cells
title_sort reactive oxygen species downregulate transient receptor potential melastatin 6 expression mediated by the elevation of mir-24-3p in renal tubular epithelial cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8393788/
https://www.ncbi.nlm.nih.gov/pubmed/34440664
http://dx.doi.org/10.3390/cells10081893
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