Ruthenium red delays the onset of cell death during oxidative stress of rat hepatocytes.
Our objective was to determine if ruthenium red protects against lethal oxidative injury of rat hepatocytes. tert-Butyl hydroperoxide, 100 mumol/L, was used to produce oxidative stress. After 2 hours of oxidative stress, cell viability was greater with than without 25 mumol/L ruthenium red (37% vs. 4.6%; P less than 0.01). Despite this cytoprotection, ruthenium red did not alter the rate or extent of glutathione depletion, malondialdehyde generation, or adenosine triphosphate depletion. In contrast, ruthenium red did retard loss of the mitochondrial membrane potential (78% vs. 42% within 30 minutes; P less than 0.01). However, the protective effect of ruthenium red could not solely be explained by preserving the mitochondrial membrane potential. Indeed, ruthenium red still improved cell survival after 2 hours of exposure to 10 mumol/L carbonyl cyanide m-chlorophenylhydrazone (CCCP), a mitochondrial uncoupler (39% vs. 13%; P less than 0.01). Cytosolic free calcium values did not change during the uncoupling of mitochondria, suggesting that the cytoprotective properties of ruthenium red cannot be explained by blocking mitochondrial calcium transport. Ruthenium red did inhibit proteolysis after 2 hours of exposure to tert-butyl hydroperoxide (434 +/- 62 vs. 242 +/- 20 nmol/10(6) cells; P = 0.016) or CCCP (236 +/- 50 vs. 99 +/- 38 nmol/10(6) cells; P = 0.04). The results indicate that ruthenium red appears to protect against hepatocellular injury by inhibiting degradative proteolytic activity. It is concluded that proteolysis may be an important mechanism contributing to lethal oxidative injury of hepatocytes.
Department of Internal Medicine, Mayo Medical School, Clinic, and Foundation, Rochester, Minnesota.
J L
JL
Groskreutz
S F
SF
Bronk
G J
GJ
Gores
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects*; D015687:Cell Hypoxia; D002462:Cell Membrane / Q000187:drug effects; D002478:Cells, Cultured; D005978:Glutathione / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000502:physiology; D011506:Proteins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012430:Ruthenium Red / Q000494:pharmacology*
D011506:Proteins; D012430:Ruthenium Red; D008315:Malondialdehyde; D000255:Adenosine Triphosphate; D005978:Glutathione; D002118:Calcium