半胱氨酸蛋白酶抑制剂减少大鼠脑缺血模型caspase-3的表达  

Cysteine Protease Inhibitors Decreased the caspase-3 Expression in Rat Brains Subjected to Transient Focal Cerebral Ischemia

作　　者：王宇卉 夏春林[2] 邵福源[3] 

机构地区：[1]解放军第100医院神经科,215007 [2]苏州大学医学院神经细胞生物学研究室 [3]第二军医大学长征医院神经科

出　　处：《中国临床神经科学》2004年第3期245-249,共5页Chinese Journal of Clinical Neurosciences

摘　　要：目的 :研究半胱氨酸蛋白酶caspase 3及calpain抑制剂干预治疗对大鼠局灶性脑缺血 /再灌注模型caspase 3表达的影响。方法 :大鼠随机分为经左侧侧脑室注射caspase 3抑制剂Ⅲ组 (DEVD组 )、calpain抑制剂Ⅰ组 (ALLN组 )、联合治疗组 (DEVD +ALLN组 )、溶剂二甲基亚砜对照组 (DMSO组 )以及左侧大脑中动脉缺血 /再灌注对照组 (MCAO组 ) ,诱导左侧大脑中动脉缺血 2h ,再灌注2 4或 48h ;再灌注 2 4h进行TTC染色观察梗死灶的形成情况 ;用TdT介导的dUTP缺口末端标记技术 (TUNEL)检测鼠脑中的神经元凋亡情况 ;并分别通过原位杂交和免疫组化技术检测鼠脑中caspase 3mRNA及活性蛋白的表达。结果 :DMSO组的各项指标与MCAO组无明显差异 ;DEVD组或ALLN组缺血侧脑中细胞凋亡数、caspase 3mRNA及活性蛋白的表达均明显减少 ,联合治疗组作用更强。结论 :caspase 3与calpain抑制剂干预治疗可减少大鼠缺血再灌注脑区神经元凋亡和caspase 3的表达 ,从而产生神经保护作用 ,并具有潜在的临床应用价值。Aim: To study the effects of cysteine protease caspase-3 and calpain inhibitor on caspase-3 expression in rat brains subjected to transient focal cerebral ischemia.Methods:Rats were randomized to receive intracerebroventricle injection of caspase-3 inhibitor Ⅲ (group DEVD), calpain inhibitor Ⅰ (group ALLN),or both (group DEVD+ALLN),vector dimethyl sulfoxide (group DMSO),or none of above (group MCAO), and then were induced to ischemia by 2 hours of left middle cerebral artery(MCA)occlusion,followed by 24 or 48 hours of recirculation. Infarct zones were confirmed by 2,3,5-triphenyltetrazolium chloride (TTC) staining at 24 hours of recirculation, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labelling (TUNEL) was used to detect neuronal apoptosis in brain sections, in situ hybridization and immunohistochemistry were performed in rats brain sections to detect caspase-3 expression at the mRNA level, and the active protein level separately.Results:Compared with MCAO rats, the apoptotic neurons, caspase-3 mRNA and active protein expressions in rats received DMSO injection were no significant different;those expressions in the ipsilateral brain of rats pretreated with caspase-3 or calpain inhibitor were remarkably decreased, in a synergic manner after the inhibitor were combined. Conclusion:Pretreatment with caspase-3 or/and calpain inhibitor might produce neuroprotection by decreasing the neuronal apoptosis and caspase-3 expression in ischemic brain, therefore caspase-3 or/and calpain inhibitors might be of potential therapeutic utility in ischemic injuries to the human brain.

关 键 词：caspase-3 表达 大鼠 CALPAIN抑制剂 左侧 再灌注 大脑中动脉缺血 RNA DMSO 侧脑室注射 

分 类 号：R743[医药卫生—神经病学与精神病学] R735[医药卫生—临床医学]