Treatment of hypoxic-ischemic brain injury in newborn rats with TPCK 3 h after hypoxia decreases caspase-9 activation and improves neuropathologic outcome.
N-Tosyl-L-phenylalanyl-chloromethyl ketone (TPCK) reduces apoptosis in vitro. Pretreatment with TPCK reduces brain injury. Would treatment after injury reduce damage? Seven-day-old rats had the right carotid artery ligated and were subjected to 2.5 h of 8% oxygen and were treated intraperitoneally 3 h after hypoxia with 10 mg/kg of TPCK or vehicle. Brain damage was measured 22 days after injury. bcl-2, bax, and cytochrome c where measured by Western blot 24 h after injury. Caspase-9 and caspase-3 activity were measured enzymatically 24 h after injury. Treatment with TPCK reduced the loss of the right hemisphere caused by injury from 27.6 +/- 2.8% SEM (vehicle, n = 56) to 19.8 +/- 2.8% (TPCK, n = 61, p < 0.05). Hypoxic ischemia increased cytosolic cytochrome c from 0.25 +/- 0.04 to 0.4 +/- 0.04 optical density (OD; p < 0.05), but TPCK had no effect (0.31 +/- 0.03 OD). TPCK reduced caspase-9 activity from 72 +/- 30 to 43 +/- 5 fluorescence units/h/mg (p < 0.05 vs. vehicle), and caspase-3 activity from 66 +/- 10 to 39 +/- 3.7 fluorescence units/h/mg (p < 0.05 vs. vehicle). Treatment with TPCK 3 h after hypoxic ischemia reduced brain infarct size. TPCK may act by reducing caspase-9 activation by cytochrome c.
Feng
Yangzheng
Y
Department of Pediatrics, University of Mississippi Medical Center, Jackson, MS 39216, USA.
LeBlanc
Michael H
MH
D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D053453:Caspase 9; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D003574:Cytochrome c Group / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004334:Drug Administration Schedule; D004789:Enzyme Activation / Q000187:drug effects; D020925:Hypoxia-Ischemia, Brain / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015842:Serine Proteinase Inhibitors / Q000008:administration & dosage*; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000008:administration & dosage*
D061945:Caspase Inhibitors; D003574:Cytochrome c Group; D018696:Neuroprotective Agents; D015842:Serine Proteinase Inhibitors; D014108:Tosylphenylalanyl Chloromethyl Ketone; C506435:Casp9 protein, rat; D053453:Caspase 9; D020169:Caspases