Estimation of Gelatinase Content in Rat Brain: Effect of Focal Ischemia

Planas, Anna M.; Solé, Sònia; Justicia, Carles; Rodrı́guez-Farré, Eduard

doi:10.1006/bbrc.2000.3881

Cited by 35 publications

(18 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To our knowledge, it has not been previously reported that MMP-2 activity and MMP-2 mRNA in newborn animals are more up-regulated after resuscitation with 100% O 2 than with ambient air. In adult animals, abnormal MMP activity has been implicated in cerebral ischemia (43). MMP-2 and MMP-9 expression levels were significantly increased soon after the ischemic onset (23,24,44).…”

Section: Discussionmentioning

confidence: 99%

Resuscitation with 100% O2 Increases Cerebral Injury in Hypoxemic Piglets

Munkeby

Børke

Björnland³

et al. 2004

Pediatr Res

120

View full text Add to dashboard Cite

Perinatal asphyxia is a major cause of immediate and postponed brain injury in the newborn. We hypothesized that resuscitation with 100% O 2 compared with ambient air is detrimental to the cerebral tissue. We assessed cerebral injury in newborn piglets that underwent global hypoxia and subsequent resuscitation with 21 or 100% O 2 by extracellular glycerol, matrix metalloproteinase (MMP) expression levels, and oxidative stress. Extracellular glycerol was sampled by cerebral microdialysis. MMP levels were analyzed in cerebral tissue by gelatin zymography, broad matrix degrading capacity, and real-time PCR. Total endogenous antioxidant capacity was measured by the oxygen radical absorbance capacity assay. Extracellular glycerol increased 50% after resuscitation with 100% O 2 compared with 21% O 2 . Total MMP activity was doubled in resuscitated animals at endpoint compared with baseline (p ϭ 0.018), and the MMP-2 activity was significantly increased in piglets that were resuscitated with 21% O 2 (p ϭ 0.003) and 100% O 2 (p ϭ 0.001) compared with baseline. MMP-2 mRNA level was 100% increased in piglets that were resuscitated with 100% O 2 as compared with 21% O 2 (p Ͻ 0.05). Oxygen radical absorbance capacity values in piglets that were resuscitated with 100% O 2 were considerably reduced compared with both baseline (p ϭ 0.001) and piglets that were resuscitated with 21% O 2 (p ϭ 0.001). In conclusion, our data show increased MMP-2 activity at both gene and protein levels, accompanied with cerebral leakage of glycerol, presumably triggered by augmented oxidative stress. Traditionally, asphyxiated newborn infants have been resuscitated using 100% oxygen in the delivery room (1). However, clinical trials have shown that room air is as efficient as pure oxygen in securing the survival of asphyxiated newborn infants (2-5). Therefore there is an ongoing debate whether to use ambient air or 100% O 2 in neonatal resuscitation (6 -9).Hyperoxia followed by reoxygenation is thought to increase the production of reactive oxygen species and disrupt the antioxidant mechanisms (10). Elevated oxidative stress can directly influence the cell cycle and additionally alter a number of significant cell functions, such as signal transduction, DNA and RNA synthesis, protein synthesis, and enzyme activation (11,12). Furthermore, hypoxic-ischemic injury and reactive oxygen species are found to trigger inflammation in the immature brain (13).Interstitial glycerol is a sensitive and reliable marker of cell damage in experimental cerebral ischemia (14 -16). Degradation of membrane phospholipids is a well-known phenomenon in acute brain injuries and is thought to underlie the disturbance of vital cellular membrane functions.Matrix metalloproteinases (MMPs) are a family of Ͼ20 zinc-and calcium-dependent endopeptidases that are involved in the remodeling of the extracellular matrix (ECM) in a variety of physiologic and pathologic conditions (17

show abstract

Section: Discussionmentioning

confidence: 99%

Resuscitation with 100% O2 Increases Cerebral Injury in Hypoxemic Piglets

Munkeby

Børke

Björnland³

et al. 2004

Pediatr Res

120

View full text Add to dashboard Cite

show abstract

“…The protein concentration in the homogenate was determined using Bradford reagent (Bio-Rad). Matrix metalloproteinase-2 and -9 in homogenates were concentrated with gelatin-sepharose beads (Gelatin Sepharose 4B, Amersham Biosciences, Piscataway, NJ, USA) and analyzed by gelatin zymography as previously reported with modifications (Planas et al, 2000). Briefly, 0.8 mg protein in 500 ml was incubated with 80 ml Gelatin-Sepharose 4B beads for 1 h at 41C with gentle rotation.…”

Section: Gelatin Zymographymentioning

confidence: 99%

Matrix Metalloproteinase-Mediated Disruption of Tight Junction Proteins in Cerebral Vessels is Reversed by Synthetic Matrix Metalloproteinase Inhibitor in Focal Ischemia in Rat

Yang

Estrada

Thompson

et al. 2006

J Cereb Blood Flow Metab

904

772

View full text Add to dashboard Cite

Matrix metalloproteinases (MMPs) disrupt the blood-brain barrier (BBB) during reperfusion. Occludin and claudins are recently described tight junction proteins (TJPs) that form the BBB. We hypothesized that the opening of the BBB was because of the degradation of TJPs by the MMPs. Spontaneously hypertensive rats had a 90 mins middle cerebral artery occlusion with reperfusion for 2, 3, or 24 h. Matrix metalloproteinases were measured by immunohistochemistry and in situ and gel zymography. Real-time polymerase chain reaction (PCR) measured mRNAs of MMP-2 and -9, furin, membrane-type MMP (MT1-MMP), occludin, and claudin-5. There was opening of the BBB in the piriform cortex after 3 h of reperfusion, and an MMP inhibitor, BB-1101 (30 mg/kg), prevented the opening. At 3 h, in situ zymograms showed gelatinase activity. Zymography and PCR showed greater increases in MMP-2 than in MMP-9. There were increased mRNA and immunohistochemistry for MT1-MMP and furin, which activate MMP-2. Claudin-5 and occludin mRNA expression decreased at 2 h in both hemispheres with fragments of both proteins seen on Western blot by 3 h on the ischemic side; treatment with BB-1101 reversed the degradation of the TJPs. Immunohistochemistry at 3 h showed fragmented TJPs within the endothelial cell clefts. By 24 h, in situ zymography showed gelatinase activity and gel zymography showed elevated levels of MMP-9. Disrupted TJPs previously seen in endothelial cells appeared in the surrounding astrocytes. Our results provide direct evidence that MMPs open the BBB by degrading TJPs and that an MMP inhibitor prevents degradation of the TJPs by MMPs.

show abstract

“…MMPs degrade components of the basal lamina, and several lines of evidence support their involvement in blood-brain barrier breakdown after ischemia (Asahi et al, 2000(Asahi et al, , 2001Fujimura et al, 1999;Gasche et al, 1999;Heo et al, 1999;Rosenberg et al, 1998). MMP-9 becomes activated in brain tissue during the first day after cerebral ischemia/reperfusion (Asahi et al, 2000;Planas et al, 2000Planas et al, , 2001Rosenberg et al, 1998Rosenberg et al, , 2001) and also after permanent ischemia (Romanic et al, 1998). A correlation between MMP-9 activity and hemorrhagic transformation of the ischemic lesion has been reported (Heo et al, 1999;Lapchack et al, 2000;Sumii et al, 2002).…”

mentioning

confidence: 94%

Neutrophil Infiltration Increases Matrix Metalloproteinase-9 in the Ischemic Brain after Occlusion/Reperfusion of the Middle Cerebral Artery in Rats

Justicia

Panés

Solé

et al. 2003

J Cereb Blood Flow Metab

Self Cite

221

191

View full text Add to dashboard Cite

Summary:Matrix metalloproteinase-9 (MMP-9) activity increases in the brain during the first day after focal ischemia and might be involved in the pathogenesis of tissue damage. We previously showed MMP-9 in the extracellular space of brain parenchyma along with neutrophil recruitment after ischemia. In the present study, we tested whether neutrophils were a direct source of enhanced MMP-9 in the ischemic brain. Neutrophil infiltration was prevented either by injecting an antibody against ICAM-1, which abrogates neutrophil adhesion to the endothelial vessel wall, or by inducing neutropenia. Onehour intraluminal middle cerebral artery occlusion with reperfusion was induced, and studies were performed at 24 hours. Circulating neutrophils expressed 95-kDa MMP-9 and dimers, and infiltrated neutrophils stained positive for MMP-9. The expression of MMP-9 (mainly 95-kDa proform and dimers and, to a lesser extent, 88-kDa form) increased in brain after ischemia/reperfusion. Treatments preventing neutrophil infiltration failed to preclude the ischemia-induced increase in 88-kDa MMP-9 form and gelatinase activity in neurons and blood vessels. However, these treatments prevented the major increase in 95-kDa MMP-9 form and dimers. We conclude that neutrophil infiltration highly contributes to enhanced MMP-9 in the ischemic brain by releasing MMP-9 proform, which might participate in the tissular inflammatory reaction.

show abstract

Estimation of Gelatinase Content in Rat Brain: Effect of Focal Ischemia

Cited by 35 publications

References 25 publications

Resuscitation with 100% O2 Increases Cerebral Injury in Hypoxemic Piglets

Resuscitation with 100% O2 Increases Cerebral Injury in Hypoxemic Piglets

Matrix Metalloproteinase-Mediated Disruption of Tight Junction Proteins in Cerebral Vessels is Reversed by Synthetic Matrix Metalloproteinase Inhibitor in Focal Ischemia in Rat

Neutrophil Infiltration Increases Matrix Metalloproteinase-9 in the Ischemic Brain after Occlusion/Reperfusion of the Middle Cerebral Artery in Rats

Contact Info

Product

Resources

About