title: Cathepsin and calpain inhibitor E64d attenuates matrix metalloproteinase-9 activity after focal cerebral ischemia in rats.
abstract: OBJECTIVE
  Matrix metalloproteinases (MMPs) and cysteine proteases (calpain and cathepsin B) play an important role in cell death and are upregulated after focal cerebral ischemia. Because there is a significant interaction between MMP-9 with calpain and cathepsin B, we investigated the role of E64d (a calpain and cathepsin B inhibitor) on MMP-9 activation in the rat focal ischemia model.
  
  
  METHODS
  Male Sprague-Dawley rats were subjected to 2 hours of middle cerebral artery occlusion by using the suture insertion method followed by 22 hours of reperfusion. In the treatment group, a single dose of E64d (5 mg/kg IP) was administrated 30 minutes before the induction of focal ischemia, whereas the nontreatment group received dimethyl sulfoxide only. The neurological deficits, infarct volumes, Evans blue extravasation, brain edema, and MMP-9 activation in the brain were determined.
  
  
  RESULTS
  Pretreatment with E64d produced a significant reduction in the cerebral infarction volume (353.1+/-19.8 versus 210.3+/-23.7 mm3) and the neurological deficits. Immunofluorescence studies showed MMP-9, calpain, and cathepsin B activation colocalized to both neurons and the neurovascular endothelial cells after ischemia, which was reduced by E64d.
  
  
  CONCLUSIONS
  These results suggest that E64d treatment provides a neuroprotective effect to rats after transient focal cerebral ischemia by inhibiting the upregulation of MMP-9.
authors.affiliation: Department of Physiology and Pharmacology, Loma Linda University School of Medicine, California, USA.
authors.forename: Tamiji
authors.initials: T
authors.lastname: Tsubokawa
authors.forename: Ihsan
authors.initials: I
authors.lastname: Solaroglu
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yatsushige
authors.forename: Julian
authors.initials: J
authors.lastname: Cahill
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Yata
authors.forename: John H
authors.initials: JH
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects; D001929:Brain Edema / Q000209:etiology / Q000473:pathology; D002154:Calpain / Q000037:antagonists & inhibitors*; D002401:Cathepsin B / Q000037:antagonists & inhibitors*; D002543:Cerebral Hemorrhage / Q000209:etiology; D015853:Cysteine Proteinase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D004353:Drug Evaluation, Preclinical; D004789:Enzyme Activation / Q000187:drug effects; D004790:Enzyme Induction / Q000187:drug effects; D005070:Evans Blue / Q000493:pharmacokinetics; D005119:Extravasation of Diagnostic and Therapeutic Materials; D020244:Infarction, Middle Cerebral Artery / Q000150:complications / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D007930:Leucine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis; D061965:Matrix Metalloproteinase Inhibitors; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors*; D018696:Neuroprotective Agents / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D011292:Premedication; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D016037:Single-Blind Method
chemical_list: D015853:Cysteine Proteinase Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D009419:Nerve Tissue Proteins; D018696:Neuroprotective Agents; D005070:Evans Blue; D002154:Calpain; D002401:Cathepsin B; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D007930:Leucine; C108192:aloxistatin