Fig. 3: Dialysis-based free hemoglobin removal reduces hemolysis-driven phospholipid peroxidation.
From: Normothermic human kidney preservation drives iron accumulation and ferroptosis
A Experimental design comparing RBC-based perfusion of deceased donor kidneys without (Group 1, n = 7) or with (Group 2, n = 4) dialysis-based free hemoglobin (fHb) removal. Perfusate fHb levels during perfusion (B), and perfusate (C) and effluent (D) fHb levels after 24 and 48 h of perfusion (Mean ± SD. Two-way ANOVA). E Total oxidized phosphatidylcholines (oxPC) in tissue as detected by targeted oxPC LC-MS/MS in control (n = 4), RBC-perfused (n = 7), and fHb removal (n = 3) groups (Mean ± SD. One-way ANOVA). F Heatmap of individual oxPC species. See Table S6 for lipid identities and their abbreviations. G Levels of arachidonic acid (AA)-derived lipid mediators in perfusate during RBC-based (n = 7) and fHb removal during RBC-based perfusion (n = 3–4) (Median with IQR). H, I Box plot of GSVA enrichment score of WP_Ferroptosis (I, n = 64 genes) and FerrDb v2 driver, suppressor and marker genes ( J, n = 313 genes) (One-way ANOVA). Boxes represent median with 25th and 75th percentile and error bars minimum to maximum. J Volcano plots of differentially expressed genes in kidneys after RBC-based perfusion or fHb removal during RBC-based perfusion, compared to control kidneys (left and middle panel). Correlation of Log2FC changes between RBC-perfused vs control kidneys and fHb removal vs control kidneys (right panel). Red highlights validated FerrDb v2 driver, suppressor, or marker genes with a protein product (n = 313). K Temporal expression dynamics in FerrDb v2 driver, suppressor, and marker genes for RBC-perfused and fHb removal groups relative to control kidneys (|Log2FC| ≥ 2 and adjusted P ≤ 0.05 for at least one of the comparisons). Two-sided Wald test with BH correction ( J, K). Association between perfusate fHb and lipid peroxidation markers after 48-h of perfusion: perfusate TBARS (MDA) (L) and tissue 4-hydroxynonenal (4HNE) (M) (Pearson’s correlation coefficient, two-sided. Shaded area indicates 95% CI). A Illustrated by Manon Zuurmond (scientific illustrator). Source data are provided as a Source Data file.
