Combination therapy with transductive anti-death FNK protein and FK506 ameliorates brain damage with focal transient ischemia in rat.
Many practical therapies have been explored as clinical applications for ischemic cerebral infarction; however, most are still insufficient to treat acute stroke. We show here a potential combination therapy in a rat focal ischemic model to improve neurological symptoms as well as to reduce infarct volumes at the maximum level. We applied protein transduction technology using artificial anti-death Bcl-xl derivative with three amino acid-substitutions (Y22F, Q26N and R165K) (FNK) protein fused with a protein-transduction-domain peptide (PTD-FNK). When PTD-FNK was administrated 1 h after initiating ischemia followed by the administration of an immunosuppressant FK506 with a 30-min time lag, infarct volumes of the total brain and cortex were markedly reduced to 27% and 14%, respectively. This procedure not only reduced the infarct volume and edema, but also markedly improved neurological symptoms. The therapeutic effect continued for at least 1 week after ischemia. FK506 inhibited the transduction of PTD-FNK in vitro, which explains the requirement of a time lag for the administration of FK506. An additional in vitro experiment showed that PTD-FNK, when administered 30 min before FK506, gave the maximal protective effect by reducing the intracellular calcium concentration. We propose that this combination therapy would provide a synergistic protective effect by both drugs, reducing adverse the effects of FK506.
Katsura
Ken-ichiro
K
Department of Internal Medicine, Nippon Medical School, Tokyo, Japan.
Takahashi
Kumiko
K
Asoh
Sadamitsu
S
Watanabe
Megumi
M
Sakurazawa
Makoto
M
Ohsawa
Ikuroh
I
Mori
Takashi
T
Igarashi
Hironaka
H
Ohkubo
Seiji
S
Katayama
Yasuo
Y
Ohta
Shigeo
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D020520:Brain Infarction / Q000188:drug therapy* / Q000378:metabolism / Q000517:prevention & control; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D004195:Disease Models, Animal; D004357:Drug Synergism; D004359:Drug Therapy, Combination; D020925:Hypoxia-Ischemia, Brain / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D007166:Immunosuppressive Agents / Q000494:pharmacology / Q000627:therapeutic use; D002546:Ischemic Attack, Transient / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D016559:Tacrolimus / Q000494:pharmacology* / Q000627:therapeutic use; D014161:Transduction, Genetic / Q000379:methods; D016896:Treatment Outcome
D007166:Immunosuppressive Agents; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C093328:Plk3 protein, mouse; D017346:Protein Serine-Threonine Kinases; D016559:Tacrolimus