title: Mild hypothermia enhanced the protective effect of protein therapy with transductive anti-death FNK protein using a rat focal transient cerebral ischemia model.
abstract: We previously reported that the protein transduction domain fused FNK (PTD-FNK) protein, which was derived from anti-apoptotic Bcl-xL protein and thereby gained higher anti-cell death activity, has a strong neuroprotective effect on rat focal brain ischemia models. The aim of this study was to investigate the effect of PTD-FNK protein and hypothermia combined therapy on cerebral infarction. Male SD rats were subjected to 120min middle cerebral artery occlusion (MCAO) with intraluminal thread. Rats were divided into 4 groups: 1) 37°C vehicle administration (37V); 2) 37°C PTD-FNK administration (37F); 3) 35°C vehicle administration (35V); and 4) 35°C PTD-FNK administration (35F). PTD-FNK protein was intravenously administered 60min after the induction of MCAO. Hypothermia (35°C) was applied during 120min MCAO. Rats were sacrificed 24h later; infarct volumes were measured, and Bax, Bcl-2, TUNEL and caspase-12 immunostaining was evaluated. There was significant infarct volume reduction in 37F, 35V, and 35F groups compared to 37V. There was also a significant difference between 37F and 35F. This suggests that hypothermia enhanced the effect of PTD-FNK. Similar results were found in neurological symptoms. Caspase-12 and TUNEL staining showed a significant difference between 37F and 35F; however, Bax and Bcl-2 staining failed to show a difference. In this study we showed an additive protective effect of hypothermia on PTD-FNK treatment, and immunohistological results showed that the protective mechanisms might involve the inhibition of apoptotic pathways through caspase-12, but not through Bcl-2.
authors.affiliation: Divisions of Neurology, Nephrology, and Rheumatology, Department of Internal Medicine, Nippon Medical School, Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Sakurazawa
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Katsura
authors.forename: Moeko
authors.initials: M
authors.lastname: Saito
authors.forename: Sadamitsu
authors.initials: S
authors.lastname: Asoh
authors.forename: Shigeo
authors.initials: S
authors.lastname: Ohta
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Katayama
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D020520:Brain Infarction / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D053454:Caspase 12 / Q000235:genetics / Q000627:therapeutic use; D004195:Disease Models, Animal; D007036:Hypothermia, Induced / Q000379:methods*; D002546:Ischemic Attack, Transient / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008297:Male; D018696:Neuroprotective Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000235:genetics / Q000494:pharmacology; D025521:Tumor Suppressor Proteins; D051020:bcl-X Protein / Q000235:genetics / Q000627:therapeutic use*; D000097868:Polo-like Kinases
chemical_list: D053454:Caspase 12; D018696:Neuroprotective Agents; D017346:Protein Serine-Threonine Kinases; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D051020:bcl-X Protein; C494813:Bcl2l1 protein, rat; C486256:PLK3 protein, human; D000097868:Polo-like Kinases; C506443:Casp12 protein, rat