Glycine protection against hypoxic injury in isolated rat proximal tubules: the role of proteases

Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association
M J TijsenJ F Wetzels

Abstract

Isolated rat proximal tubules are frequently used as a model to study hypoxic injury. Glycine is a very effective protective agent against hypoxia-induced cell injury in this model. The mechanisms involved in hypoxic renal injury and glycine protection are still debated. We have focused on the role of proteolytic enzymes. Isolated rat proximal tubules in suspension were gassed with either 95%O2/5%CO2 or 95%N2/5%CO2 to create normoxic or hypoxic conditions. Cell injury was assessed by the release of LDH. Activity of proteolytic enzymes was measured by quantifying the release of fluorescent 7-amino-4-methylcoumarin from specific substrates, which were added to tubules in suspension or to cytosolic fractions of permeabilized tubules. Fifteen minutes of hypoxia caused cell injury, which was completely prevented by glycine. Activities of serine-, aspartate-, and the calcium-dependent cysteine protease calpain were increased in these hypoxic tubules in suspension, but only calpain activity was attenuated by glycine. Cytosolic fractions obtained by digitonin-permeabilization of hypoxic (15 min) tubules showed increased proteolytic activity of all measured classes of proteases and glycine prevented these increases. In measurements perf...Continue Reading

Citations

Sep 25, 1998·Life Sciences·S M PetersJ F Wetzels
Nov 26, 2011·Canadian Journal of Physiology and Pharmacology·Israel Pérez-TorresGuadalupe Baños
Feb 13, 2001·International Journal of Experimental Pathology·P W Vanderklish, B A Bahr
Jul 23, 2004·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·Alkesh JaniCharles L Edelstein
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Jun 15, 2010·Neurochemistry International·Martina Baliova, Frantisek Jursky
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May 4, 2021·Journal of Cellular and Molecular Medicine·Sehoon ParkDong Ki Kim
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