No abstract

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-X L derivative with three amino acidsubstitutions (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. has not yet been investigated. We proceeded with experiments using a focal ischemic model in rats to prevent brain injury after transient focal ischemia by the post-administration of PTD-FNK. FK506 (Tacrolimus) is an immunosuppressant to prevent allograft rejection after organ transplantation. In addition, FK506 has a protective effect against ischemia-reperfusion injury in organs, including the heart (Nishinaka et al. 1993), liver (Garcia-Criado et al. 1997;Matsuda et al. 1998), and kidney (Sakr et al. 1992). After Sharkey and Butcher (1994) first showed that post ischemic administration of FK506 mitigates brain injury following permanent ligation of the rat middle cerebral artery (MCA), many studies have confirmed the neuroprotective effect of FK506 in various experimental models, including a focal ischemia model using a filament (Kuroda and Siesjö 1996;Arii et al. 2001) and a transient global ischemia model (Drake et al. 1996;Ide et al. 1996;Yagita et al. 1996;Katsura et al. 2003). In addition, FK506 is shown to display a neurotrophic effect (Gold et al. 1994;Lyons et al. 1994).It is estimated that there are 730 000 and 1 500 000 initial strokes each year in the United States and Japan, respectively, and that stroke is the third leading cause of death in many countries (Warlow 1998). Moreover, about 40% of bedridden aged people who are hospitalized in Japan are bedridden because of stroke (Ministry of Health, Labour and Welfare, Japan 2002), which makes stroke the leading cause of increasing medical expenses.Recently, with progress in new therapeutic approaches, a worldwide campaign was initiated ...

No abstract

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.

We have identified by imaging diagnosis the ingredients of implant materials.