Abstract
BackgroundTranscription factor Nrf2 protects from experimental acute kidney injury (AKI) and is promising to limit progression in human chronic kidney disease (CKD) by upregulating multiple antioxidant genes. We recently demonstrated that deletion of Keap1, the endogenous inhibitor of Nrf2, in T lymphocytes significantly protects from AKI. In this study, we investigated the effect of Keap1 deletion on Nrf2 mediated antioxidant response in the renal tubular epithelial cells.MethodsWe deleted Keap1 exon 2 and 3 in the renal tubular epithelial cells by crossing Ksp-Cre mice with Keap1 floxed (Keap1f/f) mice. Deletion of Keap1 gene in the kidney epithelial cells of Ksp-Keap1-/- mice and its effect on Nrf2 target gene expression was performed using PCR and real-time PCR respectively. Histological evaluation was performed on H&E stained sections. Complete blood count, serum and urine analysis were performed to assess systemic effects of defective kidney development. Student’s T test was used to determine statistical difference between the groups.ResultsKsp-Cre resulted in the deletion of Keap1 exon 2 and 3 and subsequent upregulation of Nrf2 target genes, Nqo1, Gclm and Gclc in the kidney epithelial cells of Ksp-Keap1-/- mice at baseline. Renal epithelial cell specific deletion of Keap1 in Ksp-Keap1-/- mice caused marked renal pelvic expansion and significant compression of medullary parenchyma consistent with hydronephrosis in both (3 month-old) males and females. Kidneys from 6 month-old Ksp-Keap1-/- mice showed progressive hydronephrosis. Hematological, biochemical and urinary analysis showed significantly higher red blood cell count (p = 0.04), hemoglobin (p = 0.01), hematocrit (p = 0.02), mean cell volume (p = 0.02) and mean cell hemoglobin concentration (p = 0.003) in Ksp-Keap1-/- mice in comparison to Keap1f/f mice.ConclusionsThese unexpected findings demonstrate that Keap1 deletion in renal tubular epithelial cells results in an abnormal kidney development consistent with hydronephrosis and reveals a novel Keap1 mediated signaling pathway in renal development.
Highlights
Transcription factor Nrf2 protects from experimental acute kidney injury (AKI) and is promising to limit progression in human chronic kidney disease (CKD) by upregulating multiple antioxidant genes
Nrf2 target gene Nqo1, Gclm and Gclc were significantly upregulated in kidney epithelial cell from Ksp-Keap1-/- mice compared to Keap1f/f mice (Fig. 1d), which is consistent with our previous findings in T lymphocytes (11)
MRNA levels of Nrf2 and HO-1 were found to be reduced in the kidney epithelial cells of Ksp-Keap1-/- mice
Summary
Transcription factor Nrf protects from experimental acute kidney injury (AKI) and is promising to limit progression in human chronic kidney disease (CKD) by upregulating multiple antioxidant genes. We recently demonstrated that deletion of Keap, the endogenous inhibitor of Nrf, in T lymphocytes significantly protects from AKI. We investigated the effect of Keap deletion on Nrf mediated antioxidant response in the renal tubular epithelial cells. The Keap1-Nrf cytoprotective response is critical to combat reactive oxygen species (ROS) and electrophiles generated during endogenous and exogenous stresses [1,2,3]. Keap (Kelch-like ECH-associated protein 1) is a repressor protein that regulates transcriptional activity of Nrf (nuclear factor erythroid 2-related factor 2) by. Noel et al BMC Nephrology (2016) 17:110 S No. Primer name Sequence 5′-3′
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