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Abstract: SA-PO285

Dexamethasone and Enalaprilat but Not SB525334 Exhibit Anti-Inflammatory Properties in Precision-Cut Kidney Slices (PCKSs) Obtained from Mice with Adriamycin-Induced Nephropathy

Session Information

Category: Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

  • 2000 Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

Authors

  • Pijacka, Wioletta, Sygnature Discovery Ltd, Nottingham, Nottingham, United Kingdom
  • Lincevicius, Gisele, Sygnature Discovery Ltd, Nottingham, Nottingham, United Kingdom
  • Willis, Jessica, Sygnature Discovery Ltd, Nottingham, Nottingham, United Kingdom
  • Levers, Connor Thomas, University of Nottingham, Nottingham, United Kingdom
  • Butler, Stephanie, Sygnature Discovery Ltd, Nottingham, Nottingham, United Kingdom
  • Vickers, Steven, Sygnature Discovery Ltd, Nottingham, Nottingham, United Kingdom
  • Cheetham, Sharon, Sygnature Discovery Ltd, Nottingham, Nottingham, United Kingdom
Background

Precision cut kidney slices (PCKS) provide a valuable step between in vitro and in vivo testing. PCKS obtained from rodents exposed to adriamycin exhibit specific pathological features associated with glomerular and tubular injury, inflammation and eventual fibrosis. In this study, we have evaluated the effects of dexamethasone, enalaprilat, and SB525334 on inflammation and fibrosis in PCKS from kidney slices from adriamycin-treated mice.

Methods

Male mice were dosed with adriamycin (11.25 mg/kg, iv) to induce nephropathy. On Day 7 mice were humanely sacrificed and kidneys were cut at 250µm on a vibratome. Slices were cultured in Williams Medium E media.
Tissue viability was evaluated at 0h to 96h by LDH assay and qPCR (versus control). Independent slices were treated with SB525334 (1µM), Dexamethasone (1µM) and enalaprilat (10µM) for 24 to 96h. Pro-fibrotic (Fn1, Col1a, Col3a, PAI1), pro-inflammatory genes (IL-6, TNFα, IL-1β and CCL2) and genes indicative of glomerular and tubular injury (nephrin, KIM-1 and WT1) were determined.

Results

PCKS were viable for 96h. All pro-inflammatory and pro-fibrotic gene expression were significantly increased from 24h onwards, p<0.001. Nephrin, WT1 expression decreased, and KIM-1 increased at 24h and remained elevated, p<0.001.
SB525334 decreased the expression of all pro-fibrotic genes, p<0.001 but not the pro-inflammatory markers (NS) and increased nephrin and KIM-1 (p<0.05, p<0.001) but had no effect on WT1.
Dexamethasone significantly reduced TNFα, IL-6, IL-1β, CCL2 (p<0.001) as well as Fn1, Col1a, Col3a and PAI1 (p<0.001). Dexamethasone increased WT1 and reduced KIM-1 (p<0.05, p<0.001) but unchanged nephrin.
Enalaprilat effects were limited, it reduced IL-1β (p<0.01) and increased WT1 (p<0.05).

Conclusion

Dexamethasone and enalaprilat produced a reduction in inflammation and fibrosis (dexamethasone only) leading to protection of the functional unit of the kidney. In contrast, SB525334 effects were limited to fibrosis but the effects on kidney health were inconclusive. Taken together, inflammation plays a crucial role in adriamycin-induced nephropathy and PCKS present a reliable platform to test novel anti-inflammatory and anti-fibrotic therapies.

Funding

  • Commercial Support – Sygnature Discovery