Cathepsin L在大鼠短暂性前脑缺血中的作用  

The role of Cathepsin L in transient forebrain ischemia in rats

作　　者：彭旭 张创 张智搏 孙李晴 王婷 唐敏 唐璐 周颖 

机构地区：[1]长沙市第一医院神经内科,长沙410005

出　　处：《中国医师杂志》2016年第3期375-379,共5页Journal of Chinese Physician

基　　金：湖南省医药卫生科研计划课题项目（B2012-135）

摘　　要：目的 探讨Cathepsin L在大鼠短暂性前脑缺血中的作用.方法 选取周龄为10-12周的清洁级健康雄性Sprague-Dawley （SD）大鼠60只,体重（280 ±20）g,依据随机数字表法将大鼠分为假手术组（Sham组）10只;脑缺血再灌注组（IRI组）25只;Cathepsin L抑制剂Z-FY-DMK干预组（CLI组）25只;IRI组、CLI组大鼠分别按3、6、12及24h四个时间点每个时间点分为四个亚组.采用改良longa线栓法制作大鼠大脑中动脉缺血再灌注损伤（MCAO）模型,CLI组于术前30 min侧脑室穿刺注入Z-FY-DMK（20 μg/μl）＆#215;5μl,Sham组及IRI组于术前30 min侧脑室注入浓度为10 ml/L的二甲基亚砜（DMSO）5μl.对大鼠运用Longa＆#39;s 5级标准评分法评分;2,3,5-氯化三苯基四氮唑（TTC）染色检测各组大鼠24h时间点相对脑梗死体积;Western blotting法检测半暗带区相应时间点Cathepsin L的蛋白表达变化.结果 IRI组Cathepsin L在大鼠脑缺血再灌注后3、6、12 h呈上升趋势,12 h达到高峰,24 h下降,均高于Sham组（P＜0.05）;CLI组各相应时间点Cathepsin L蛋白的表达较IRI组表达减少（P＜0.05）.结论 Cathepsin L可能参与了大鼠短暂性前脑缺血再灌注损伤后细胞凋亡.Objective To investigate the role of Cathepsin L in transient forebrain ischemia in rats.Methods Seventy healthy male Sprague-Dawley Rats （10 -12 weeks old,260 - 300 g） were chosen.Based on the random number table method,the rats were randomly divided into sham-operated control group （Sham group,n =10）,ischemia-reperfusion group （IRI group,n =25）,and Z-FY-DMK intervention group （CLI group,n =25）.Rats in groups IRI and CLI were randomly divided into 3 h,6 h,12 h,and 24 h subgroups.Modified transient middle cerebral artery occlusion was made as Longa described.The intervention groups were injected intracerebroventricularly Z-FY-DMK （20 μg/μl×5 μl） preoperative 30min prior to surgery.Groups Sham and IRI were injected intracerebroventricularly dimethyl sulfoxide （DMSO） 5 μl （10 ml/L） at the same time.Longa＇s five level＇s grading was used to evaluate the neural function defect.2,3,5-Triphenyltetrazolium chloride （TTC） staining method was used to measure infarct volume at 24 h.Western blot was used to detect the expression of Cathepsin L.Results Western blot found there was little visible Cathepsin L protein expression in ischemic cerebral cortex preoptic,Cathepsin L protein expression in IRI group after cerebral ischemia-reperfusion upward at 3 h,6 h,and 12 h;it reached peak at 12 h,and declined at 24 h,was still higher than the sham group （P 〈0.01）.Cathepsin L protein expression in the CLI group was reduced compared to the IRI group （P 〈 0.05）.Conclusions Cathepsin L may be involved in neuronal apoptosis.

关 键 词：组织蛋白酶类/拮抗剂和抑制剂/代谢 脑缺血/代谢/病理学/药物疗法 细胞凋亡/药物作用 

分 类 号：R743.31[医药卫生—神经病学与精神病学]