Caspase inhibitor z-DEVD-fmk attenuates calpain and necrotic cell death in vitro and after traumatic brain injury.

Abstract

In studies designed to evaluate the therapeutic window for treatment of traumatic brain injury, the caspase 3 inhibitor z-DEVD-fmk improved neurologic function and reduced lesion volumes when administered at 1 but not at 4, 8, or 24 hours after injury. Moreover, neither caspase 3 nor PARP, a caspase 3 substrate, were cleaved in injured, untreated cortex from 1 to 72 hours after injury. Few cortical neurons expressed active caspase 3 or were TUNEL positive from 6 to 24 hours after injury, and TUNEL staining was primarily Type I (necrotic). Nissl staining revealed extensive neuronal necrosis in the injured cortex from 6 to 24 hours after impact. Considered together, these data suggested that z-DEVD-fmk may reduce neuronal necrosis, so we used an in vitro model of necrotic cell death induced by maitotoxin to test this further and explore the potential mechanism(s) involved. Z-DEVD-fmk (1 nM-100 microM) significantly attenuated maitotoxin induced neuronal cell death and markedly reduced expression of the 145 kD calpain-mediated alpha-spectrin breakdown product after maitotoxin injury. Neither the 120 kD caspase-mediated alpha-spectrin cleavage product nor cathepsin B were expressed after maitotoxin injury. In a cell free assay, z-DEVD-fmk reduced hydrolysis of casein by purified calpain I. Finally, z-DEVD-fmk reduced expression of the 145 kD calpain-mediated alpha-spectrin cleavage fragment after traumatic brain injury in vivo. These data suggest that neuroprotection by z-DEVD-fmk may, in part, reflect inhibition of calpain-related necrotic cell death.

• Authors: Susan M Knoblach, Daniel A Alroy, Maria Nikolaeva, Ibolja Cernak, Bogdan A Stoica, Alan I Faden
• Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism (J Cereb Blood Flow Metab) Vol. 24 Issue 10 Pg. 1119-32 (Oct 2004) ISSN: 0271-678X [Print] United States
• PMID: 15529012 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Caspase Inhibitors
• Cysteine Proteinase Inhibitors
• Oligopeptides
• benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone
• Calpain
• Casp3 protein, mouse
• Caspase 3
• Caspases

Topics

• Animals
• Brain Injuries (drug therapy, metabolism, pathology)
• Calpain (antagonists & inhibitors, metabolism)
• Caspase 3
• Caspase Inhibitors
• Caspases (metabolism)
• Cell-Free System
• Cysteine Proteinase Inhibitors (pharmacology)
• In Vitro Techniques
• Male
• Mice
• Mice, Inbred C57BL
• Necrosis
• Nerve Degeneration (drug therapy, metabolism, pathology)
• Oligopeptides (pharmacology)