蛋白激酶抑制剂H-7降低局灶性脑缺血半暗带和核心区半胱氨酸蛋白酶的活化

目的 研究蛋白激酶抑制剂H-7对局灶性脑缺血半暗带和核心区半胱氨酸蛋白酶Calpain和Caspase-3活性的影响.方法 采用动脉腔内插线法建立大鼠局灶性脑缺血模型,在缺血前15min经脑室给予H-7(125μg/大鼠),测定缺血1h再灌注23h(R23h)时,半暗带和核心区Calpain和Caspase-3的活性、Calpastatin和微管相关蛋白-2(MAP-2)的含量及梗死体积.结果 H-7明显降低R23h时半暗带和核心区μ-和m-Calpain及Caspase-3的活性,升高核心区Calpastatin的含量及半暗带和核心区MAP-2的含量,缩小梗死体积.结论 H-7通过抑制半暗带和核心区Calpain和Caspase-3的活性,降低局灶性脑缺血损伤.

Abstract：

Objective To investigate the effects of protein kinase inhibitor H-7 on the activation of calpain and caspase-3 in penumbra and core after focal cerebral ischemia in rats. Methods Rats received 1h focal cerebral ischemia by intraluminal filament. H-7 ( 125 μg/rat) was administered intracerebroventricularly 15 min before ischemia. The activities of calpain and caspase-3, the levels of calpastatin and microtubule-associated protein-2 ( MAP-2 ) , and the infarct volume were assessed by casein zymography,fluorometry, Western blot analysis,and staining the brain sections with 2,3 ,5-tripheny-ltetrazolium chlorides,respectively. Results Compared with ischemic control, H-7 markedly reduced the activities of μ-and m-calpain, and caspase-3 , increased the levels of MAP-2 in penumbra and core, and enhanced the levels of calpastatin in core. Moreover, animals treated with H-7 showed a significant reduction in infarct volume. Conclusions These data demonstrate the protection of H-7 against focal cerebral ischemia through inhibiting the activation of calpain and caspase-3.

H-7 reduces cysteine protease activation in the rat model of focal cerebral ischemia

Objective To investigate the effects of protein kinase inhibitor H-7 on the activation of calpain and caspase-3 in penumbra and core after focal cerebral ischemia in rats. Methods Rats received 1h focal cerebral ischemia by intraluminal filament. H-7 ( 125 μg/rat) was administered intracerebroventricularly 15 min before ischemia. The activities of calpain and caspase-3, the levels of calpastatin and microtubule-associated protein-2 ( MAP-2 ) , and the infarct volume were assessed by casein zymography,fluorometry, Western blot analysis,and staining the brain sections with 2,3 ,5-tripheny-ltetrazolium chlorides,respectively. Results Compared with ischemic control, H-7 markedly reduced the activities of μ-and m-calpain, and caspase-3 , increased the levels of MAP-2 in penumbra and core, and enhanced the levels of calpastatin in core. Moreover, animals treated with H-7 showed a significant reduction in infarct volume. Conclusions These data demonstrate the protection of H-7 against focal cerebral ischemia through inhibiting the activation of calpain and caspase-3.