Remote ischemic preconditioning causes transient cell cycle arrest and renal protection by a NF-κB–dependent Sema5B pathway
Jan Rossaint, … , Hermann Pavenstädt, Alexander Zarbock
Jan Rossaint, … , Hermann Pavenstädt, Alexander Zarbock
Published June 21, 2022
Citation Information: JCI Insight. 2022;7(14):e158523. https://doi.org/10.1172/jci.insight.158523.
View: Text | PDF
Research Article Immunology Nephrology

Remote ischemic preconditioning causes transient cell cycle arrest and renal protection by a NF-κB–dependent Sema5B pathway

Abstract

Acute kidney injury increases morbidity and mortality, and previous studies have shown that remote ischemic preconditioning (RIPC) reduces the risk of acute kidney injury after cardiac surgery. RIPC increases urinary high mobility group box protein-1 (HMGB1) levels in patients, and this correlates with kidney protection. Here, we show that RIPC reduces renal ischemia-reperfusion injury and improves kidney function in mice. Mechanistically, RIPC increases HMGB1 levels in the plasma and urine, and HMGB1 binds to TLR4 on renal tubular epithelial cells, inducing transcriptomic modulation of renal tubular epithelial cells and providing renal protection, whereas TLR4 activation on nonrenal cells was shown to contribute to renal injury. This protection is mediated by activation of induction of AMPKα and NF-κB; this induction contributes to the upregulation of Sema5b, which triggers a transient, protective G1 cell cycle arrest. In cardiac surgery patients at high risk for postoperative acute kidney injury, increased HMGB1 and Sema5b levels after RIPC were associated with renal protection after surgery. The results may help to develop future clinical treatment options for acute kidney injury.

Authors

Jan Rossaint, Melanie Meersch, Katharina Thomas, Sina Mersmann, Martin Lehmann, Jennifer Skupski, Tobias Tekath, Peter Rosenberger, John A. Kellum, Hermann Pavenstädt, Alexander Zarbock

×

Figure 5

HMGB1 induces TLR4-dependent cell cycle arrest in renal tubular epithelial cells.

Options: View larger image (or click on image) Download as PowerPoint
HMGB1 induces TLR4-dependent cell cycle arrest in renal tubular epitheli...
After induction of general anesthesia, renal IRI was induced in WT mice by clamping of the renal pedicles for 32 minutes. Some mice received a cyclin-dependent kinase (CDK) inhibitor (PD0332991, 100 mg/kg) or a control before inducing renal IRI. Twenty-four hours after the surgery, mice were sacrificed. (A) The recruitment of neutrophils (PMNs) into the kidney was analyzed by flow cytometry (n = 4). (B) Serum creatinine levels were measured by a photometric assay (n = 4). (C) Isolated murine renal tubular epithelial cells were treated with control, HMGB1 (0.1 μg/mL), a TLR4 inhibitor (TAK-242), or HMGB1 together with TLR4 inhibitor in vitro for 24 hours. Cell cycle analysis was performed by measuring cellular DNA content by flow cytometry (n = 6). (D) Isolated murine renal tubular epithelial cells were treated with different concentrations of HMGB1 in presence or absence of of TLR4 inhibitor (TAK-242), and cell apoptosis was analyzed by propidium iodide/annexin V staining and flow cytometry (n = 6). (E) Isolated murine renal tubular epithelial cells were treated with HMBG1 0.1 μg/mL, HMGB1 10 μg/mL, or HMGB1 10 μg/mL plus TLR4 inhibitor (TAK-242) for 1 hour. The proportion of cells in G0/G1 phase was analyzed by measuring cellular DNA content by flow cytometry (n = 6). (F and G) Isolated murine renal tubular epithelial cells from WT and TLR4fl/fl/Ksp-Cre+/T mice were treated with HMGB1 and TIMP-2 (F) and IGFBP7 (G) released into the supernatant were analyzed by ELISA (n = 4). (H and I) Cell cycle arrest in renal tubular epithelial cells was analyzed by flow cytometry (n = 4). (J–L) The levels of the chemokines CXCL1 (J), CXCL2 (K), and IL-6 (L) in kidney tissue homogenisates were analyzed by ELISAs (n = 4). Mann-Whitney U test (J–L) and 1-way ANOVA followed by Bonferroni testing (A–I) were used for statistical analysis; *P < 0.05.