Accumulation of nonesterified fatty acids causes the sustained energetic deficit in kidney proximal tubules after hypoxia-reoxygenation.
Kidney proximal tubules exhibit decreased ATP and reduced, but not absent, mitochondrial membrane potential (Deltapsi(m)) during reoxygenation after severe hypoxia. This energetic deficit, which plays a pivotal role in overall cellular recovery, cannot be explained by loss of mitochondrial membrane integrity, decreased electron transport, or compromised F1F0-ATPase and adenine nucleotide translocase activities. Addition of oleate to permeabilized tubules produced concentration-dependent decreases of Deltapsi(m) measured by safranin O uptake (threshold for oleate = 0.25 microM, 1.6 nmol/mg protein; maximal effect = 4 microM, 26 nmol/mg) that were reversed by delipidated BSA (dBSA). Cell nonesterified fatty acid (NEFA) levels increased from <1 to 17.4 nmol/mg protein during 60- min hypoxia and remained elevated at 7.6 nmol/mg after 60 min reoxygenation, at which time ATP had recovered to only 10% of control values. Safranin O uptake in reoxygenated tubules, which was decreased 85% after 60-min hypoxia, was normalized by dBSA, which improved ATP synthesis as well. dBSA also almost completely normalized Deltapsi(m) when the duration of hypoxia was increased to 120 min. In intact tubules, the protective substrate combination of alpha-ketoglutarate + malate (alpha-KG/MAL) increased ATP three- to fourfold, limited NEFA accumulation during hypoxia by 50%, and lowered NEFA during reoxygenation. Notably, dBSA also improved ATP recovery when added to intact tubules during reoxygenation and was additive to the effect of alpha-KG/MAL. We conclude that NEFA overload is the primary cause of energetic failure of reoxygenated proximal tubules and lowering NEFA substantially contributes to the benefit from supplementation with alpha-KG/MAL.
Feldkamp
Thorsten
T
Nephrology Division, Department of Internal Medicine, Rm. 1560, MSRB II, University of Michigan Medical Center, Ann Arbor, MI 48109-0676, USA.
Kribben
Andreas
A
Roeser
Nancy F
NF
Senter
Ruth A
RA
Weinberg
Joel M
JM
D000255:Adenosine Triphosphate / Q000096:biosynthesis*; D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D045304:Cytochromes c / Q000494:pharmacology; D005230:Fatty Acids, Nonesterified / Q000378:metabolism* / Q000493:pharmacokinetics; D005260:Female; D066298:In Vitro Techniques; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D008928:Mitochondria / Q000502:physiology*; D010619:Phenazines / Q000378:metabolism; D011817:Rabbits; D015427:Reperfusion Injury / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D005230:Fatty Acids, Nonesterified; D010619:Phenazines; D012710:Serum Albumin, Bovine; D000255:Adenosine Triphosphate; D045304:Cytochromes c; C009195:safranine T