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The protein-bound uremic toxin p-cresyl-sulfate promotes intracellular ROS production and lipid peroxidation in 3T3-L1 adipose cells

Abstract : Patients with chronic kidney disease (CKD) often exhibit increased level of oxidative stress that contribute to the deterioration of renal function and uremic complications. White adipose tissue (WAT) has been recognized as a major site of production of radical oxygen species (ROS) in the context of metabolic diseases. This study was designed to decipher whether the protein bound uremic toxin p-cresyl-sulfate (p-CS) could contribute to ROS production in WAT and promote oxidative stress. Mouse 3T3-L1 adipocytes were incubated for 2 h in culture medium containing 212 μM p-CS, a concentration chosen to mimic levels encountered in end stage renal disease patients or KCl as a control and intracellular ROS production was measured using the fluorescent probe 5-6-carboxy-2',7'-dichlorodihydrofluorescein diacetate. Oxidative insult was estimated by the measurement of malondialdehyde (MDA) content and glutathione content. The effects of probenecid (1 mM) a potent inhibitor of organic anion transporter, apocynin (1 mM) an inhibitor of NADPH oxidase or common antioxidants such as α-tocopherol (2.5 μM), ascorbate (200 μM), and N-acetylcysteine (500 μM) were further evaluated. p-CS triggered a striking increase in ROS production (+228%, p \textless 0.01), in MDA content (+214%, p \textless 0.005) and a decrease in glutathione (-47%, P \textless 0.01). Pre-treatment of cells with probenecid, apocynin or antioxidants prevented the p-CS induced ROS production and oxidative insults. These results suggest that in uremic state, the intracellular accumulation of p-CS in adipose cells could contribute, through an activation of NADPH oxidase, to the redox imbalance often reported in CKD patients.
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Submitted on : Friday, November 26, 2021 - 5:35:19 PM
Last modification on : Thursday, December 23, 2021 - 1:43:39 PM
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L. Koppe, M. L. Croze, E. B. Monteiro, B. Benoit, E. Bres, et al.. The protein-bound uremic toxin p-cresyl-sulfate promotes intracellular ROS production and lipid peroxidation in 3T3-L1 adipose cells. Biochimie, Elsevier, 2021, 189, pp.137-143. ⟨10.1016/j.biochi.2021.06.020⟩. ⟨inserm-03452097⟩



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