Loss of receptor interacting protein kinases 3 and caspase-8 augments intrinsic apoptosis in tubular epithelial cell and promote kidney ischaemia-reperfusion injury.

Abstract

BACKGROUND:

Ischaemia-reperfusion injury (IRI) is associated with programmed cell death that promotes inflammation and organ dysfunction. Necroptosis is mediated by members of receptor interacting protein kinases (RIPK1/3). Inhibition of RIPK1/3 provides a pro-survival benefit in kidney IRI. Caspase-8 initiates apoptosis and contributes to IRI. We studied whether inhibiting both RIPK3 and caspase-8 would provide an additional benefit in kidney IRI.

METHODS:

A clamp was applied to the left kidney pedicle for 45 min followed by right kidney nephrectomy. Kidney and serum from wild type, RIPK3-/- , and RIPK3-/- caspase-8-/- double knockout (DKO) mice were collected post-IRI for assessment of injury. Tubular epithelial cells (TEC) isolated from wild type, RIPK3-/- , and DKO mice were treated with interferons-γ and interleukin-1β to induce apoptotic death.

RESULTS:

Kidney IRI of DKO mice did not show improvement over RIPK3-/- mice. We have found that DKO triggered 'intrinsic' apoptosis in TEC in response to interleukin-1β and interferons-γ. Up-regulation of the B-cell lymphoma 2 (Bcl-2)-associated death promoter, the Bcl-2-homologous antagonist killer and Bcl-2-associated X protein and enhanced activation of caspase-3 and 9 were found in DKO TEC. TEC infected with Murine cytomegalovirus that encodes multiple cell death inhibitors resist to death.

CONCLUSION:

We show that the deletion of both RIPK3 and caspase-8 does not provide additive benefit in IRI or TEC death and may enhance injury by up-regulation of intrinsic apoptosis. This suggests blocking multiple death pathways may be required for the prevention of kidney IRI clinically.

• Authors: Baekjun Sung, Ye Su, Jifu Jiang, Patrick Mcleod, Weihua Liu, Aaron Haig, Douglas R Green, Zhu-Xu Zhang, Anthony M Jevnikar
• Journal: Nephrology (Carlton, Vic.) (Nephrology (Carlton)) Vol. 24 Issue 6 Pg. 661-669 (Jun 2019) ISSN: 1440-1797 [Electronic] Australia
• PMID: 30175514 (Publication Type: Journal Article)
• Copyright: © 2018 Asian Pacific Society of Nephrology.

Chemical References

• IFNG protein, mouse
• IL1B protein, mouse
• Interleukin-1beta
• Interferon-gamma
• Receptor-Interacting Protein Serine-Threonine Kinases
• Ripk3 protein, mouse
• Casp8 protein, mouse
• Caspase 8

Topics

• Animals
• Apoptosis (drug effects)
• Caspase 8 (genetics, metabolism)
• Disease Models, Animal
• Epithelial Cells (drug effects, enzymology, pathology)
• Interferon-gamma (pharmacology)
• Interleukin-1beta (pharmacology)
• Kidney Diseases (enzymology, genetics, pathology)
• Kidney Tubules (drug effects, enzymology, pathology)
• Male
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• NIH 3T3 Cells
• Necroptosis
• Receptor-Interacting Protein Serine-Threonine Kinases (deficiency, genetics)
• Reperfusion Injury (enzymology, genetics, pathology)
• Signal Transduction