Renal mouse proximal tubular cells are more susceptible than MDCK cells to chemical anoxia.
To elucidate the mechanisms responsible for the resistance of continuous cell lines to anoxic injury, we have compared the effects of ATP depletion induced by chemical anoxia on primary cultures of mouse proximal tubular (MPT) cells and on Madin-Darby canine kidney (MDCK) cells. Inhibition of ATP production by cyanide and 2-deoxyglucose (CN+DOG) in the absence of dextrose reduced cell ATP content to < 5% of control values in MPT cells and caused progressive deterioration in mitochondrial function as well as loss of cell viability in these cells. Cell free fatty acid (FFA) content rose from 4.3 +/- 0.9 to 23.7 +/- 2.0 micrograms/mg of total lipid weight after 4 h of CN + DOG (P < 0.05). The mitochondrial injury and cell death induced by CN + DOG in MPT cells was ameliorated by the addition of fatty acid-free bovine albumin to the cell medium, which reduced cell FFA content during chemical anoxia from 25.0 +/- 3.0 to 10.4 +/- 2.0 micrograms/mg (P < 0.05). The phospholipase A2 (PLA2) inhibitor, mepacrine, also resulted in functional protection and reduction of cell FFA content from 20.2 +/- 2.3 to 15.9 +/- 1.7 micrograms/mg (P < 0.05). These data suggest a role for phospholipase activation and accumulation of toxic lipid metabolites in the pathophysiology of MPT cell injury. We then compared cell injury induced by CN + DOG in MPT and MDCK cells. Despite comparable reduction in cell ATP content in the two cell types, injury was far more severe in MPT than MDCK cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Sheridan
A M
AM
Renal Section, Evans Memorial Department of Clinical Research, University Hospital, Boston, Massachusetts 02118.
Schwartz
J H
JH
Kroshian
V M
VM
Tercyak
A M
AM
Laraia
J
J
Masino
S
S
Lieberthal
W
W
D000255:Adenosine Triphosphate / Q000172:deficiency / Q000378:metabolism; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003486:Cyanides; D003847:Deoxyglucose; D004198:Disease Susceptibility; D004285:Dogs; D005227:Fatty Acids / Q000378:metabolism; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D006801:Humans; D000860:Hypoxia / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology*; D007231:Infant, Newborn; D007687:Kidney Tubules, Proximal / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008928:Mitochondria / Q000502:physiology; D011796:Quinacrine / Q000494:pharmacology; D012709:Serum Albumin / Q000494:pharmacology
D003486:Cyanides; D005227:Fatty Acids; D005230:Fatty Acids, Nonesterified; D012709:Serum Albumin; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D011796:Quinacrine