Indoxyl Sulfate Contributes to Uremic Sarcopenia by Inducing Apoptosis in Myoblasts

Author(s)

Cardoso Rodrigues, Gabriela Gomes ^[1] ; Delle, Humberto ^[1] ; Oliveira Brito, Rodrigo Barbosa ^[1] ; Cardoso, Vinicius Oliveira ^[2] ; Santos Fernandes, Kristianne Porta ^[3] ; Mesquita-Ferrari, Raquel Agnelli ^[3] ; Cunha, Regiane Stafim ^[4] ; Marques Stinghen, Andrea Emilia ^[4] ; Dalboni, Maria Aparecida ^[1] ; Barreto, Fellype Carvalho ^[5]

Affiliation

^[1] Univ Nove Julho, Postgrad Program Med, Sao Paulo - Brazil

^[2] Univ Nove Julho, Postgrad Program Rehabil Sci, Sao Paulo - Brazil

^[3] Univ Nove Julho, Postgrad Program Biophoton Appl Hlth Sci, Sao Paulo - Brazil

^[4] Univ Fed Parana, Basic Pathol Dept, Expt Nephrol Lab, Curitiba, Parana - Brazil

^[5] Univ Fed Parana, Dept Internal Med, Nephrol Serv, Curitiba, Parana - Brazil

• Document type: Journal article
• Source: ARCHIVES OF MEDICAL RESEARCH; v. 51, n. 1, p. 21-29, JAN 2020.
• Web of Science Citations: 0

Abstract

Objetive. Uremic sarcopenia is a complication of chronic kidney disease, particularly in its later stages, which leads to musculoskeletal disability. Uremic toxins have been linked to the pathogenesis of several manifestations of uremic syndrome. We sought to investigate whether indoxyl sulphate (IS), a protein-bound uremic toxin, is implicated in the development of uremic sarcopenia. Material and Methods. Myoblasts were exposed to IS at normal (0.6 mg/L, IS0.6), uremic (53 mg/L, IS53) or maximum uremic (236 mg/L, IS236) concentrations for 24, 48 and 72 h. Cell viability was evaluated by MTT assay and by 7-aminoactinomycin D staining. ROS generation and apoptosis were evaluated by flow cytometry. MyoD and myogenin mRNA expression was evaluated by qRT-PCR and myosin heavy chain expression by immunocytochemistry. Results. Myoblast viability was reduced by IS236 in a time-dependent pattern (p < 0.05; 84.4, 68.0, and 63.6%). ROS production was significantly higher (p < 0.05) in cells exposed to IS53 and IS236 compared to control (untreated cells). The apoptosis rate was significantly higher in cells treated with IS53 and IS236 than in control after 48h (p < 0.05; 4.7 +/- 0.1% and 4.6 +/- 0.3% vs. 3.1 +/- 0.1%, respectively) and 72h (p < 0.05; 9.6 +/- 1.1% and 10.4 +/- 0.3% vs. 3.1 +/- 0.7%, respectively). No effect was observed on MyoD, myogenin, myosin heavy chain expression, and markers of myoblast differentiation at any IS concentration tested or time-point experiment. Conclusions. These data indicate that IS has direct toxic effects on myoblast by decreasing its viability and increasing cell apoptosis. IS may be a potential target for treating uremic sarcopenia. (C) 2020 IMSS. Published by Elsevier Inc. (AU)

• FAPESP's process: 14/14192-1 - Effect of vitamin 25 (OH) d on the expression of Toll-like receptor and inflammatory mediators in the uremic environment
• Grantee: Maria Aparecida Dalboni
• Support Opportunities: Regular Research Grants