Inhibition of caspase-1-like activity by Ac-Tyr-Val-Ala-Asp-chloromethyl ketone induces long-lasting neuroprotection in cerebral ischemia through apoptosis reduction and decrease of proinflammatory cytokines.
Broad spectrum caspase inhibitors have been found to reduce neurodegeneration caused by cerebral ischemia. We studied whether blockade of group I caspases, mainly caspase-1, using the inhibitor Ac-YVAD.cmk reduced infarct volume and produced prolonged neuroprotection. Ac-YVAD.cmk (300 ng/rat) was injected intracerebroventricularly 10 min after permanent middle cerebral artery occlusion in the rat. Drug treatment induced a significant reduction of infarct volume not only 24 hr after ischemia (total damage, percentage of hemisphere volume: control, 41.1 +/- 2.3%; treated, 26.5 +/- 2.1%; p < 0.05) but also 6 d later (total damage: control, 30.6 +/- 2.2%; treated, 23.0 +/- 2.2%; p < 0.05). Ac-YVAD. cmk treatment resulted in a reduction not only of caspase-1 (control, 100 +/- 20.3%; treated, 3.4 +/- 10.4%; p < 0.01) but also of caspase-3 (control, 100 +/- 30.3%; treated, 13.2 +/- 9.5%; p < 0.05) activity at 24 hr and led to a parallel decrease of apoptosis as measured by nucleosome quantitation (control, 100 +/- 11.8%; treated, 47 +/- 5.9%; p < 0.05). Six days after treatment no differences in these parameters could be detected between control and treated animals. Likewise, brain levels of the proinflammatory cytokines IL-1beta and TNF-alpha were reduced at 24 hr (39.5 +/- 23.7 and 51.9 +/- 10.3% of control, respectively) but not at 6 d. Other cytokines, IL-10, MCP-1, MIP-2, and the gaseous mediator nitric oxide, were not modified by the treatment. These findings indicate that blockade of caspase-1-like activity induces a long-lasting neuroprotective effect that, in our experimental conditions, takes place in the early stages of damage progression. Finally, this effect is achieved by interfering with both apoptotic and inflammatory mechanisms.
Rabuffetti
M
M
Schering-Plough Research Institute, and Department of Biotechnology, San Raffaele Science Park, Milan, Italy.
Sciorati
C
C
Tarozzo
G
G
Clementi
E
E
Manfredi
A A
AA
Beltramo
M
M
D000590:Amino Acid Chloromethyl Ketones / Q000008:administration & dosage / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D061945:Caspase Inhibitors; D002540:Cerebral Cortex / Q000187:drug effects / Q000276:immunology / Q000473:pathology*; D002544:Cerebral Infarction / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D018932:Chemokine CCL2 / Q000096:biosynthesis; D054426:Chemokine CXCL2; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis*; D020287:In Situ Nick-End Labeling; D007249:Inflammation; D007276:Injections, Intraventricular; D007375:Interleukin-1 / Q000096:biosynthesis; D016753:Interleukin-10 / Q000096:biosynthesis; D002546:Ischemic Attack, Transient / Q000473:pathology / Q000503:physiopathology*; D008297:Male; D015846:Monokines / Q000096:biosynthesis; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009707:Nucleosomes / Q000187:drug effects / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D018932:Chemokine CCL2; D054426:Chemokine CXCL2; D015853:Cysteine Proteinase Inhibitors; D016207:Cytokines; D007375:Interleukin-1; D015846:Monokines; C098738:N-acetyl-tyrosyl-valyl-alanyl-aspartyl chloromethyl ketone; D018696:Neuroprotective Agents; D009707:Nucleosomes; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10