The mechanism of Heme oxygenase-1 regulated by Hypoxia-inducible factor-1 in acute kidney injury
Objective This study investigates the role and mechanism of hypoxia-inducible factor-1(HIF-1)in acute kidney injury(AKI)induced by renal ischemia-reperfusion(IR).Methods A total of 20 C57 mice,aged 6-8 weeks and weighing 20-25 g.This group included 10 wild-type(WT)mice and 10 mice with tubular epithelial cell-specific knockdown of HIF-1(cKO).The mice were randomly assigned to either a sham operation(Sham)group or an IR group,with 5 mice in each subgroup.1％pentobarbital sodium was administered intravenously for anesthesia.In the surgical procedure,the renal arteries and veins were ligated for 60 minutes and then released.Kidneys were harvested 24 hours later to establish the ischemia-reperfusion model.Renal tubular damage was evaluated using hematoxylin-eosin staining(HE).Glutathione(GSH),malondialdehyde(MDA),and iron levels in kidney tissue were determined by en-zyme-linked immunosorbent assay(ELISA).Western blotting was employed to assess heme oxygenase-1(HO-1)protein expression and proteins related to iron-induced cell death.Immunohistochemistry(IHC)was utilized to detect the expression of HIF-1,HO-1,and proteins associated with iron-induced cell death.All data were presented as mean±standard deviation,and group comparisons were made using an inde-pendent t-test.Results HE staining indicated that the IR group of WT mice exhibited more severe renal tubular epithelial cell damage compared to the Sham group(9.78±1.07 vs.2.43±0.93,t=5.438,P＜0.05).In the cKO mice,cell damage in the IR group was more severe than in the WT-IR group after HIF-1 knockdown(28.59±2.57 vs.9.78±1.07,t=3.348,P＜0.05),showing statistically significant differences.Western blotting results revealed higher expression of HIF-1 and HO-1 in the IR group of WT mice compared to the Sham group(HIF-1:0.996±0.068 vs.0.634±0.072,HO-1:0.734±0.153 vs.0.592±0.097,both P＜0.05).Immunohistochemistry findings showed that in the WT-IR group,both HIF-1 and HO-1 expressions were significantly higher compared to the cKO-IR group(HIF-1:8.52±0.21 vs.4.35±0.16,HO-1:8.53±0.22 vs.3.43±0.18,both P＜0.05).Conclusion In the renal ische-mia-reperfusion model,HIF-1 expression is upregulated.Knockdown of HIF-1 exacerbates kidney injury,suggesting that HIF-1 may play a protective role by regulating HO-1 to reduce cell death due to iron over-load.
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