Humanin is a novel neuroprotective agent against stroke.
OBJECTIVE
Humanin (HN) is a 24-amino acid peptide best known for its ability to protect neurons from damage caused by Alzheimer disease-related proteins. This study examines the neuroprotective effects of HNG (a potent form of HN) on focal cerebral ischemia/reperfusion injury in mice.


METHODS
Mice underwent middle cerebral artery occlusion for 75 minutes followed by 24-hour reperfusion. Mice were pretreated with 0.1 microg HNG (intracerebroventricularly) 30 minutes before ischemia; posttreated at 0, 2, 4, and 6 hours after ischemia; or pretreated with 1 microg HNG (intraperitoneally) 1 hour before ischemia. Neurological deficits and cerebral infarct volume were evaluated. Neuronal apoptosis and activated poly(ADP-ribose) polymerase expression were measured by TUNEL and Western blot analysis, respectively. Activated ERKs were examined by Western blot analysis.


RESULTS
Pretreatment with 0.1 microg HNG (intracerebroventricularly) 30 minutes before ischemia reduced cerebral infarct volume from 56.2+/-3.0% to 26.1+/-1.4% (P<0.01). HNG posttreatment after 4 hours of reperfusion reduced cerebral infarct volume to 45.6+/-2.6% (P<0.05). Pretreatment with 1 microg HNG (intraperitoneally) 1 hour before ischemia or posttreatment after 2 hours of reperfusion reduced cerebral infarct volume significantly. HNG also significantly improved neurological function and inhibited both neuronal apoptosis as well as poly(ADP-ribose) polymerase activation. A significant decrease of phospho-ERK was observed in mice treated with HNG, whereas phospho-JNK and phospho-p38 levels were not altered.


CONCLUSIONS
Our results demonstrate that HNG protects against cerebral ischemia/reperfusion injury in mice. HNG offers neuroprotection in vivo at least in part by inhibiting ERK activation. These findings suggest a potential therapeutic role for HNG in the treatment of stroke.
Xu
Xingshun
X
Department of Pharmacology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN 37614, USA.
Chua
Chu C
CC
Gao
Jinping
J
Hamdy
Ronald C
RC
Chua
Balvin H L
BH
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001812:Blood-Brain Barrier; D002545:Brain Ischemia / Q000201:enzymology / Q000209:etiology / Q000517:prevention & control*; D004353:Drug Evaluation, Preclinical; D004789:Enzyme Activation / Q000187:drug effects; D020244:Infarction, Middle Cerebral Artery / Q000150:complications; D007276:Injections, Intraventricular; D048031:JNK Mitogen-Activated Protein Kinases / Q000032:analysis; D008297:Male; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D009069:Movement Disorders / Q000209:etiology / Q000517:prevention & control; D009474:Neurons / Q000187:drug effects / Q000473:pathology; D018696:Neuroprotective Agents / Q000493:pharmacokinetics / Q000627:therapeutic use*; D010455:Peptides / Q000493:pharmacokinetics / Q000627:therapeutic use*; D011897:Random Allocation; D015427:Reperfusion Injury / Q000201:enzymology / Q000473:pathology / Q000517:prevention & control; D020521:Stroke / Q000201:enzymology / Q000209:etiology / Q000517:prevention & control*; D048051:p38 Mitogen-Activated Protein Kinases / Q000032:analysis
C450880:Gly(14)-Humanin; D018696:Neuroprotective Agents; D010455:Peptides; D048031:JNK Mitogen-Activated Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases