Caspase Inhibition after Neonatal Ischemia in the Rat Brain

Caspase-3 has been identified as a key protease in the execution of apoptosis and appears to be an important downstream event after hypoxia-ischemia in the immature brain. The efficacy of a pan-caspase inhibitor, boc-aspartyl-(Ome)-fluoromethyl-ketone (BAF), was tested in a model of unilateral focal ischemia with reperfusion in 7-day-old rats. The BAF inhibitor was given intraperitoneally 5 minutes before reperfusion via the carotid artery. This procedure reduced the activity of caspase-3 by 79% but did not induce a significant reduction in infarct volume (23.8 ± 7.5% versus 30.1 ± 6.4%). Animals were distributed in two populations. One population exhibited an infarct, whereas the other appeared to be fully protected. BAF-treated animals exhibiting an infarct mostly displayed necrotic cell death, whereas apoptotic nuclei were observed in untreated or vehicle-treated animals. Repeated dose of BAF (5 minutes before and 9 hours after reperfusion) did not also provide benefit after neonatal ischemia, although a general trend to reduce lesion was observed (20.5 ± 3.7% versus 34.4 ± 5.9%). These findings raise critical questions about the use of peptide ketone apoptotic inhibitors in improving histopathologic outcomes after neonatal stroke.

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Caspase Inhibition Prevents the Increase in Caspase-3, -2, -8 and -9 Activity and Apoptosis in the Cold Ischemic Mouse Kidney

American Journal of Transplantation ◽

10.1111/j.1600-6143.2004.00498.x ◽

2004 ◽

Vol 4 (8) ◽

pp. 1246-1254 ◽

Cited By ~ 50

Author(s):

Alkesh Jani ◽

Danica Ljubanovic ◽

Sarah Faubel ◽

Jun Kim ◽

Ron Mischak ◽

...

Keyword(s):

Caspase 3 ◽

Mouse Kidney ◽

Caspase Inhibition

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Caspase inhibition in apoptotic T cells triggers necrotic cell death depending on the cell type and the proapoptotic stimulus

Journal of Cellular Biochemistry ◽

10.1002/jcb.20670 ◽

2006 ◽

Vol 97 (6) ◽

pp. 1350-1361 ◽

Cited By ~ 20

Author(s):

Carsten Scheller ◽

Johanna Knöferle ◽

Anett Ullrich ◽

Johannes Prottengeier ◽

Tomas Racek ◽

...

Keyword(s):

T Cells ◽

Cell Death ◽

Necrotic Cell Death ◽

Cell Type ◽

Necrotic Cell ◽

Caspase Inhibition

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Biphasic changes in the levels of poly(ADP‐ribose) polymerase‐1 and caspase 3 in the immature brain following hypoxia–ischemia

International Journal of Developmental Neuroscience ◽

10.1016/j.ijdevneu.2005.08.002 ◽

2005 ◽

Vol 23 (8) ◽

pp. 673-686 ◽

Cited By ~ 22

Author(s):

Shannon S. Martin ◽

J. Regino Perez‐Polo ◽

Kristin M. Noppens ◽

Marjorie R. Grafe

Keyword(s):

Caspase 3 ◽

Immature Brain ◽

Hypoxia Ischemia

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The Protective Effect of Ceramide in Immature Rat Brain Hypoxia—Ischemia Involves Up-Regulation of BCL-2 and Reduction of TUNEL-Positive Cells

Journal of Cerebral Blood Flow & Metabolism ◽

10.1097/00004647-200101000-00005 ◽

2001 ◽

Vol 21 (1) ◽

pp. 34-40 ◽

Cited By ~ 54

Author(s):

Yong Chen ◽

Irene Ginis ◽

John M. Hallenbeck

Keyword(s):

Infarct Volume ◽

Treated Group ◽

Protective Role ◽

Necrosis Factor Alpha ◽

Immature Rat ◽

Tnf Α ◽

Hypoxia Ischemia ◽

Old Rats ◽

Factor Alpha ◽

The Right

Preconditioning brain with tumor necrosis factor alpha (TNF-α) can induce tolerance to experimental hypoxia and stroke and ceramide is a downstream messenger in the TNF-α signaling pathway. A hypoxic-ischemic (HI) insult in the immature rat injures brain primarily through apoptosis. Apoptosis is regulated by Bcl-2 family proteins. The authors explored whether ceramide protects against HI in the immature rat, and whether Bcl-2 family protein expression is involved. Hypoxia-ischemia was produced in seven-day-old rats by ligating the right carotid artery, followed by 2 hours of 8% oxygen exposure. Thirty minutes after HI, C2-ceramide (150 μg/kg) was injected intraventricularly. Infarct volume was measured 5 days later. C2-ceramide reduced HI-induced brain damage by 45% to 65% compared with HI/dimethyl sulfoxide (DMSO) (vehicle control) or HI only groups. In separate experiments, brains of sham-operated control and HI only animals and animals subjected to HI plus C2-ceramide or DMSO infusion were sampled 6 hours, 24 hours, and 5 days after treatments and analyzed for Bcl-2, Bcl-xl, and Bax expression (Western blotting), and apoptosis (TUNEL assay). Augmented Bcl-2 and Bcl-xl levels in the C2-ceramide treated group were associated with a significant decrease in TUNEL-positive cells. The results support a protective role for ceramide in neonatal HI.

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Concurrent Assessment of Calpain and Caspase-3 Activation after Oxygen–Glucose Deprivation in Primary Septo-Hippocampal Cultures

Journal of Cerebral Blood Flow & Metabolism ◽

10.1097/00004647-200111000-00004 ◽

2001 ◽

Vol 21 (11) ◽

pp. 1281-1294 ◽

Cited By ~ 47

Author(s):

Jennifer K. Newcomb-Fernandez ◽

Xiurong Zhao ◽

Brian R. Pike ◽

Kevin K. W. Wang ◽

Andreas Kampfl ◽

...

Keyword(s):

Cell Death ◽

Caspase 3 ◽

Apoptotic Cell ◽

Glucose Deprivation ◽

Apoptotic Cell Death ◽

Oxygen Glucose Deprivation ◽

Necrotic Cell Death ◽

Necrotic Cell ◽

Lactate Dehydrogenase Release ◽

Hippocampal Cultures

The contributions of calpain and caspase-3 to apoptosis and necrosis after central nervous system (CNS) trauma are relatively unexplored. No study has examined concurrent activation of calpain and caspase-3 in necrotic or apoptotic cell death after any CNS insult. Experiments used a model of oxygen–glucose deprivation (OGD) in primary septo-hippocampal cultures and assessed cell viability, occurrence of apoptotic and necrotic cell death phenotypes, and protease activation. Immunoblots using an antibody detecting calpain and caspase-3 proteolysis of α-spectrin showed greater accumulation of calpain-mediated breakdown products (BDPs) compared with caspase-3–mediated BDPs. Administration of calpain and caspase-3 inhibitors confirmed that activation of these proteases contributed to cell death, as inferred by lactate dehydrogenase release. Oxygen–glucose deprivation resulted in expression of apoptotic and necrotic cell death phenotypes, especially in neurons. Immunocytochemical studies of calpain and caspase-3 activation in apoptotic cells indicated that these proteases are almost always concurrently activated during apoptosis. These data demonstrate that calpain and caspase-3 activation is associated with expression of apoptotic cell death phenotypes after OGD, and that calpain activation, in combination with caspase-3 activation, could contribute to the expression of apoptotic cell death by assisting in the degradation of important cellular proteins.

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Inhibition of ?-ketoglutarate dehydrogenase complex promotes cytochromec release from mitochondria, caspase-3 activation, and necrotic cell death

Journal of Neuroscience Research ◽

10.1002/jnr.10756 ◽

2003 ◽

Vol 74 (2) ◽

pp. 309-317 ◽

Cited By ~ 50

Author(s):

Hsueh-Meei Huang ◽

Hsiu-Chong Ou ◽

Hui Xu ◽

Huan-Lian Chen ◽

Corinne Fowler ◽

...

Keyword(s):

Cell Death ◽

Caspase 3 ◽

Necrotic Cell Death ◽

Necrotic Cell ◽

Dehydrogenase Complex ◽

Ketoglutarate Dehydrogenase

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Caspase-3 Activity and Carbonyl Cyanide m-Chlorophenylhydrazone-induced Apoptosis in HL-60 cells

Alternatives to Laboratory Animals ◽

10.1177/026119290102900313 ◽

2001 ◽

Vol 29 (3) ◽

pp. 243-249 ◽

Cited By ~ 8

Author(s):

Petr Mlejnek

Keyword(s):

Cell Death ◽

Dna Cleavage ◽

Caspase 3 ◽

Chromatin Condensation ◽

Experimental System ◽

Apoptotic Bodies ◽

Nucleosomal Dna ◽

Carbonyl Cyanide ◽

Induced Apoptosis ◽

Fluoromethyl Ketone

The role of caspase proteases in carbonyl cyanide m-chlorophenylhydrazone (CCCP)-induced apoptosis of human promyelocytic HL-60 cells was examined. Treatment of HL-60 cells with micromolar concentrations of CCCP resulted in cell death, with typical apoptotic features such as chromatin condensation, formation of apoptotic bodies, nucleosomal fragmentation of DNA and a distinct increase in caspase-3 activity. The results, however, indicated that full caspase-3 inhibition by the selective inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethyl ketone (Z-DEVD-FMK) did not prevent cell death, nor did it affect the manifestation of apoptotic hallmarks, including apoptotic bodies formation and nucleosomal DNA fragmentation. The only distinct effect that Z-DEVD-FMK exhibited was to retard the disruption of the plasma membrane. We therefore assume that caspase-3 activity itself is not essential for the manifestation of apoptotic features mentioned above. Similarly, the pan-specific caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone (Z-VAD-FMK) did not prevent cell death. On the contrary, Z-VAD-FMK completely prevented DNA cleavage and apoptotic body formation, but it failed to completely counteract chromatin condensation. Thus, in the presence of Z-VAD-FMK, application of CCCP concentrations that otherwise induced apoptosis, resulted in the appearance of two morphologically different groups of dead cells with intact DNA. The first group included cells with necrotic-like nuclear morphology, and therefore could be taken as being “truly” necrotic in nature, because they had intact DNA. The cells of the second group formed small single-spherical nuclei with condensed chromatin. In spite of having intact DNA, they could not be taken as “truly” necrotic cells. It is evident that in the experimental system, caspase proteases play an essential role in the formation of apoptotic bodies and in the cleavage of nucleosomal DNA, but not in the condensation of chromatin. Therefore, it is likely that the choice between cell death modalities is not solely a matter of the caspase proteases present.

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Neuroprotective Effect of DAHP via Antiapoptosis in Cerebral Ischemia

Behavioural Neurology ◽

10.1155/2018/5050469 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Yanhua Qin ◽

Weiming Hu ◽

Yang Yang ◽

Zhiying Hu ◽

Weiyun Li ◽

...

Keyword(s):

Nitric Oxide ◽

Brain Injury ◽

Cerebral Ischemia ◽

Caspase 3 ◽

Focal Cerebral Ischemia ◽

Infarct Volume ◽

Neuroprotective Effect ◽

Ischemia Reperfusion ◽

Tunel Staining ◽

Cox 2

Aberrant production of nitric oxide following inducible nitric oxide synthase (iNOS) expression has been implicated in cell death and contributes to ischemic brain injury. Tetrahydrobiopterin (BH4) is an essential cofactor of NOS activity. Herein, we evaluated antiapoptotic and anti-inflammatory effects of diamino-6-hydroxypyrimidine (DAHP), a guanosine 5′-triphosphate cyclohydrolase 1 (GTPCH1) inhibitor on focal cerebral ischemia-reperfusion injury by middle cerebral artery occlusion and reperfusion (MCAO) and investigated the underlying mechanism. Sprague-Dawley rats were divided into five groups. Experimental groups were subjected to 1.5 h transient MCAO. T2-weighted imaging was performed to evaluate brain edema lesions in the stroke rats. Infarct volume was estimated by 2,3,5-triphenyltetrazolium chloride (TTC) staining after 24 h reperfusion. Western blotting and immunohistochemistry were performed to detect iNOS, caspase-3, Bcl-2, COX-2, and TNF-α protein expressions. Apoptosis was determined by TUNEL staining. T2 hyperintensity changes were observed in primary ischemic region. DAHP pretreatment significantly suppressed iNOS overexpression, caspase-3, and TNF-α. There was also attenuation of neuronal apoptosis with decrement in proteins Bcl-2 and COX-2 expressions. On the basis of our results, we hypothesize DAHP to have a neuroprotective function against focal cerebral ischemia and might attenuate brain injury by decreasing reactive oxygen species (ROS) production, subsequently inhibiting apoptosis.

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