Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways
It is hypothesized that the uremic toxin indoxyl sulfate (IS) plays a role in the pathogenesis of renal anemia. To further explore that hypothesis, we examined the effects of IS on reactive oxygen species (ROS) production, levels of reduced glutathione (GSH), and erythrocyte death (eryptosis) in red...
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| Format: | Article |
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MDPI AG
2018-07-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/10/7/280 |
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| author | Gabriela Ferreira Dias Natalia Borges Bonan Thiago Maass Steiner Sara Soares Tozoni Silvia Rodrigues Lia Sumie Nakao Viktoriya Kuntsevich Roberto Pecoits Filho Peter Kotanko Andréa N. Moreno-Amaral |
| author_facet | Gabriela Ferreira Dias Natalia Borges Bonan Thiago Maass Steiner Sara Soares Tozoni Silvia Rodrigues Lia Sumie Nakao Viktoriya Kuntsevich Roberto Pecoits Filho Peter Kotanko Andréa N. Moreno-Amaral |
| author_sort | Gabriela Ferreira Dias |
| collection | DOAJ |
| description | It is hypothesized that the uremic toxin indoxyl sulfate (IS) plays a role in the pathogenesis of renal anemia. To further explore that hypothesis, we examined the effects of IS on reactive oxygen species (ROS) production, levels of reduced glutathione (GSH), and erythrocyte death (eryptosis) in red blood cells (RBC) from healthy controls (CON-RBC) and hemodialyzed patients (HD-RBC), respectively. RBC were incubated either in either TRIS-Glc-BSA buffer or IS at concentrations of 0.01, 0.09, and 0.17 mM, respectively. We measured ROS generation (expressed as % of DCFH-DA positive RBC), eryptosis (expressed as % of annexin-V positive RBC), and GSH levels after 6, 12, and 24 h. When incubated in buffer, ROS production was approximately seven-fold higher at all time points HD-RBC when compared to CON-RBC. Incubation with IS increased ROS production in CON-RBS dose-dependently up to 10-fold. Eryptosis in buffer-incubated HD-RBC was up to seven-fold higher as compared to COB-RBC. Incubation of CON-RBC with IS increased the eryptosis rate dose-dependently up to 6-fold. Pretreatment of CON-RBC with the organic anion transporter 2 (OAT2) specific inhibitor ketoprofen or with NADPH oxidase inhibitor diphenyleneiodonium-Cl blunted the IS effect on both ROS production and eryptosis induction. While GSH levels in HD-RBC were reduced when compared to CON-RBC, they were not affected by IS incubation. In summary, IS increases ROS generation and eryptosis in CON-RBC by an activity dependent of the IS influx through OAT2, and NADPH oxidase activity-dependent, and a GSH-independent mechanism. These findings lend support to a putative role of IS in the pathogenesis of renal anemia. |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-13T08:50:13Z |
| publishDate | 2018-07-01 |
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| spelling | doaj.art-bb5a17bce6814fea9bc3207431108b6b2022-12-22T02:53:31ZengMDPI AGToxins2072-66512018-07-0110728010.3390/toxins10070280toxins10070280Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent PathwaysGabriela Ferreira Dias0Natalia Borges Bonan1Thiago Maass Steiner2Sara Soares Tozoni3Silvia Rodrigues4Lia Sumie Nakao5Viktoriya Kuntsevich6Roberto Pecoits Filho7Peter Kotanko8Andréa N. Moreno-Amaral9Graduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná; Curitiba-PR 80215-901, BrasilienGraduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná; Curitiba-PR 80215-901, BrasilienGraduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná; Curitiba-PR 80215-901, BrasilienGraduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná; Curitiba-PR 80215-901, BrasilienDepartment of Basic Pathology, Federal University of Paraná, Curitiba-PR 80060-000, BrazilDepartment of Basic Pathology, Federal University of Paraná, Curitiba-PR 80060-000, BrazilRenal Research Institute, New York, NY 10065, USAGraduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná; Curitiba-PR 80215-901, BrasilienRenal Research Institute, New York, NY 10065, USAGraduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná; Curitiba-PR 80215-901, BrasilienIt is hypothesized that the uremic toxin indoxyl sulfate (IS) plays a role in the pathogenesis of renal anemia. To further explore that hypothesis, we examined the effects of IS on reactive oxygen species (ROS) production, levels of reduced glutathione (GSH), and erythrocyte death (eryptosis) in red blood cells (RBC) from healthy controls (CON-RBC) and hemodialyzed patients (HD-RBC), respectively. RBC were incubated either in either TRIS-Glc-BSA buffer or IS at concentrations of 0.01, 0.09, and 0.17 mM, respectively. We measured ROS generation (expressed as % of DCFH-DA positive RBC), eryptosis (expressed as % of annexin-V positive RBC), and GSH levels after 6, 12, and 24 h. When incubated in buffer, ROS production was approximately seven-fold higher at all time points HD-RBC when compared to CON-RBC. Incubation with IS increased ROS production in CON-RBS dose-dependently up to 10-fold. Eryptosis in buffer-incubated HD-RBC was up to seven-fold higher as compared to COB-RBC. Incubation of CON-RBC with IS increased the eryptosis rate dose-dependently up to 6-fold. Pretreatment of CON-RBC with the organic anion transporter 2 (OAT2) specific inhibitor ketoprofen or with NADPH oxidase inhibitor diphenyleneiodonium-Cl blunted the IS effect on both ROS production and eryptosis induction. While GSH levels in HD-RBC were reduced when compared to CON-RBC, they were not affected by IS incubation. In summary, IS increases ROS generation and eryptosis in CON-RBC by an activity dependent of the IS influx through OAT2, and NADPH oxidase activity-dependent, and a GSH-independent mechanism. These findings lend support to a putative role of IS in the pathogenesis of renal anemia.http://www.mdpi.com/2072-6651/10/7/280chronic kidney diseaseindoxyl sulfateeryptosisoxidative stress |
| spellingShingle | Gabriela Ferreira Dias Natalia Borges Bonan Thiago Maass Steiner Sara Soares Tozoni Silvia Rodrigues Lia Sumie Nakao Viktoriya Kuntsevich Roberto Pecoits Filho Peter Kotanko Andréa N. Moreno-Amaral Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways Toxins chronic kidney disease indoxyl sulfate eryptosis oxidative stress |
| title | Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways |
| title_full | Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways |
| title_fullStr | Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways |
| title_full_unstemmed | Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways |
| title_short | Indoxyl Sulfate, a Uremic Toxin, Stimulates Reactive Oxygen Species Production and Erythrocyte Cell Death Supposedly by an Organic Anion Transporter 2 (OAT2) and NADPH Oxidase Activity-Dependent Pathways |
| title_sort | indoxyl sulfate a uremic toxin stimulates reactive oxygen species production and erythrocyte cell death supposedly by an organic anion transporter 2 oat2 and nadph oxidase activity dependent pathways |
| topic | chronic kidney disease indoxyl sulfate eryptosis oxidative stress |
| url | http://www.mdpi.com/2072-6651/10/7/280 |
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