Rhabdomyolysis aggravates renal iron accumulation and acute kidney injury in a humanized mouse model of sickle cell disease

Figures & data

Table 1. Oligonucleotide primer sequences.

Gene	Sequence	Ascension	Length (bp)
Ferritin LC	Forward 5′-GCCAATTCGCGGAAGAAGT-3′	NC_000073.7	101
	Reverse 5′-CGCGTTCCGTTAACTCGTATTT-3′
Ferritin HC	Forward 5′-GCACTGCACTTGGAAAAGAGT-3′	NC_000085.7	104
	Reverse 5′-ACTCATCCCACGTTTCCGAT-3′
β-actin	Forward 5′-GCTTTGCACATGCCGGA-3′	NC_000071.7	94
	Reverse 5′-GGTGACTATAGCCTTCTTTTGTGTC-3′

Figure 1. Glycerol-induced rhabdo is apparent in 5 h and aggravated in SS mice. Five hours after glycerol injection, plasma, and urine samples were harvested to measure rhabdo markers in AA and SS mice. (A) Plasma creatine kinase and (B) urinary myoglobin in control and rhabdo AA and SS mice (one-way ANOVA, with Holm-Šídák’s posthoc test).

Figure 2. Rhabdo increases plasma heme levels in SS mice. Plasma heme levels in control and rhabdo AA and SS mice (one-way ANOVA, with Holm-Šídák’s posthoc test).

Figure 3. Rhabdo increases kidney iron accumulation and upregulates ferritin heavy chain in SS mice. (A) Kidney sections were stained with Prussian blue to detect iron deposition in AA and SS mice, (B) scatter plots summarizing Prussian blue stain density in the mice, (C-D) relative mRNA expression of ferritin light chain (LC) and heavy chain (HC) in the kidneys of control and rhabdo AA and SS mice (one-way ANOVA, with Holm-Šídák’s posthoc test); scale bar = 50 µm.

Figure 4. Rhabdo induces oxidative stress in SS mice. (A) Plasma H₂O₂ and (B) urinary 8-OHdG in control and rhabdo AA and SS mice (one-way ANOVA, with Holm-Šídák’s posthoc test).

Figure 5. Rhabdo-induced oxidative stress aggravates AKI parameters in SS mice. (A) GFR, (B) urinary NGAL, (C) plasma L-FABP, and (D) plasma IL-18 in control AA and SS, rhabdo AA and SS, and TEMPOL + rhabdo SS groups (one-way ANOVA, with Holm-Šídák’s posthoc test).

Figure 6. Rhabdo-induced oxidative stress causes significant renal tubular damage in SS mice. (A) Representative H&E-stained kidney sections and (B) box & whiskers plot of tubular injury score in control AA (n = 4) and SS (n = 5), rhabdo AA (n = 5) and SS (n = 6), and TEMPOL + rhabdo SS (n = 5) groups. (one-way ANOVA, with Holm-Šídák’s posthoc test); scale bar = 50 µm.