Kallikrein gene transfer protects against ischemic stroke by promoting glial cell migration and inhibiting apoptosis.
Kallikrein/kinin has been shown to protect against ischemia/reperfusion-induced myocardial infarction and apoptosis. In the present study, we examined the potential neuroprotective action of kallikrein gene transfer in cerebral ischemia. Adult, male Sprague-Dawley rats were subjected to a 1-hour occlusion of the middle cerebral artery followed by intracerebroventricular injection of adenovirus harboring either the human tissue kallikrein gene or the luciferase gene. Kallikrein gene transfer significantly reduced ischemia-induced locomotor deficit scores and cerebral infarction after cerebral ischemia injury. Expression of recombinant human tissue kallikrein was identified and localized in monocytes/macrophages of rat ischemic brain by double immunostaining. Morphological analyses showed that kallikrein gene transfer enhanced the survival and migration of glial cells into the ischemic penumbra and core, as identified by immunostaining with glial fibrillary acidic protein. Cerebral ischemia markedly increased apoptotic cells, and kallikrein gene delivery reduced apoptosis to near-normal levels as seen in sham control rats. In primary cultured glial cells, kinin stimulated cell migration but inhibited hypoxia/reoxygenation-induced apoptosis in a dose-dependent manner. The effects of kinin on both migration and apoptosis were abolished by icatibant, a bradykinin B2 receptor antagonist. Enhanced cell survival after kallikrein gene transfer occurred in conjunction with markedly increased cerebral nitric oxide levels and phospho-Akt and Bcl-2 levels but reduced caspase-3 activation, NAD(P)H oxidase activity, and superoxide production. These results indicate that kallikrein gene transfer provides neuroprotection against cerebral ischemia injury by enhancing glial cell survival and migration and inhibiting apoptosis through suppression of oxidative stress and activation of the Akt-Bcl-2 signaling pathway.
Xia
Chun-Fang
CF
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
Yin
Hang
H
Borlongan
Cesar V
CV
Chao
Lee
L
Chao
Julie
J
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002545:Brain Ischemia / Q000175:diagnosis / Q000378:metabolism / Q000517:prevention & control*; D020169:Caspases / Q000378:metabolism; D002465:Cell Movement; D002470:Cell Survival; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000473:pathology; D002544:Cerebral Infarction / Q000473:pathology / Q000517:prevention & control; D005822:Genetic Vectors; D007610:Kallikreins / Q000235:genetics*; D008297:Male; D019255:NADPH Oxidases / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D018696:Neuroprotective Agents; D009569:Nitric Oxide / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D020521:Stroke / Q000175:diagnosis / Q000473:pathology / Q000517:prevention & control*; D013481:Superoxides / Q000378:metabolism; D020840:Tissue Kallikreins / Q000032:analysis / Q000378:metabolism
D018696:Neuroprotective Agents; D013481:Superoxides; D009569:Nitric Oxide; D019255:NADPH Oxidases; D007610:Kallikreins; D020840:Tissue Kallikreins; D020169:Caspases