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Physiological Reports

IF: 2.2
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Mitigating hemoglobin‐induced nephropathy: ApoHb‐hp protection of podocytes

Published:1 November 2024 DOI: 10.14814/phy2.70132 PMID: 39578364
Daniela Lucas, Carlos Munoz, Quintin O'Boyle, Ivan S. Pires, Andre F. Palmer, Pedro Cabrales

Abstract

This study investigates hemoglobin (Hb)-induced kidney injury and the protective role of the ApoHemoglobin-Haptoglobin (ApoHb-Hp) complex against heme and Hb damage. Hb facilitates oxygen (O2) delivery but poses challenges outside red blood cells (RBCs) due to toxic Hb and heme mechanisms. These are managed by binding to serum proteins like Haptoglobin (Hp) and Hemopexin (Hpx). During hemolysis, depletion of Hp and Hpx leaves tissues vulnerable to Hb and heme. To address this, we developed the ApoHb-Hp complex, based on Apohemoglobin, which is produced by removing heme from Hb, conjugated with Hp. This complex acts as a dual scavenger for Hb and heme, preventing tissue damage. Our findings demonstrate that ApoHb-Hp significantly protects MPC5 podocytes from Hb-induced damage. Fluorescent staining showed a higher percentage of nephrin-positive cells in the ApoHb-Hp group, and MTT assays revealed enhanced cell viability compared to Hb alone. Additionally, ApoHb-Hp reduced reactive oxygen species (ROS) production, with the Hb group exhibiting significantly elevated ROS levels. The ApoHb-Hp complex mitigated the depletion of protective mechanisms, as shown by significant increases in superoxide dismutase (SOD) and glutathione (GSH). Moreover, ApoHb-Hp treatment reduced the activation of the NLRP3 inflammasome signaling pathway and inflammatory cytokines IL-1β and IL-18. These findings underscore the therapeutic potential of ApoHb-Hp in mitigating Hb-induced renal damage by preserving podocyte viability and reducing oxidative stress. Overall, ApoHb-Hp maintained protective mechanisms depleted otherwise by Hb. These findings highlight ApoHb-Hp's potential as a therapeutic agent against Hb-induced renal damage, offering insights into its mechanisms and implications for treating conditions involving hemolysis.

Substances (2)

Materials
Procduct Name CAS Molecular Formula Supplier Price
Thiazolyl Blue 298-93-1 C18H16BrN5S 415 suppliers $7.00-$1596.00
2′,7′-Dichlorodihydrofluorescein Diacetate 4091-99-0 C24H16Cl2O7 192 suppliers $33.00-$1947.00

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