缺血后处理对脑缺血再灌注损伤后Caspase-3、Caspase-9激活和细胞色素C释放的影响  

Effects of Ischemic Postprocessing on Activation of Caspase-3 and Caspase-9 and Release of Cytochrome C in Reperfusion Injury after Cerebral Ischemia

作　　者：邢变枝[1] 陈晖[2] 王磊[2] 刘林[2] 

机构地区：[1]武汉大学人民医院放射科,武汉430060 [2]武汉大学人民医院泌尿外科,武汉430060

出　　处：《临床误诊误治》2013年第8期76-79,共4页Clinical Misdiagnosis & Mistherapy

基　　金：国家自然科学基金资助项目(30901552;30901494)

摘　　要：目的探讨缺血后处理对大鼠脑缺血再灌注损伤后Caspase-3、Caspase-9激活和细胞色素C(Cyt C)释放的影响。方法选择成年健康雄性SD大鼠60只,随机分为假手术组、缺血再灌注组、缺血后处理组,每组20只。线栓法建立大脑中动脉闭塞再灌注模型。缺血再灌注24 h后留取大脑组织,采用原位末端标记法检测脑内神经细胞凋亡情况,采用酶活性测定、Western blot和免疫组化等方法检测Caspase-3、Caspase-9及Cyt C含量。结果缺血再灌注组、缺血后处理组较假手术组细胞凋亡增多,差异均有统计学意义(P<0.05);缺血后处理组较缺血再灌注组凋亡细胞减少,差异有统计学意义(P<0.05)。缺血再灌注组、缺血后处理组Caspase-3、Caspase-9活性较假手术组高,缺血后处理组活性显著低于缺血再灌注组,差异均有统计学意义(P<0.05)。缺血再灌注组、缺血后处理组Caspase-3、Caspase-9前体蛋白proCaspase-3、proCaspase-9含量较假手术组减少(P<0.05),而缺血后处理组含量高于缺血再灌注组,差异有统计学意义(P<0.05)。在胞浆中假手术组无明显Cyt C释放,缺血再灌注组、缺血后处理组胞浆内Cyt C释放较假手术组增多,差异有统计学意义(P<0.05);缺血后处理组Cyt C释放较缺血再灌注组明显减少,差异有统计学意义(P<0.05)。线粒体中缺血再灌注组、缺血后处理组Cyt C较假手术组外漏明显,而缺血后处理组Cyt C漏出较缺血再灌注组明显减少,差异均有统计学意义(P<0.05)。结论缺血后处理可以减轻脑缺血再灌注损伤后神经细胞凋亡,抑制Caspase-3、Caspase-9的激活和Cyt C的释放。Objective To investigate the effect of ischemic postprocessing on activation of Caspase-3 and Caspase-9 and release of cytochrome C in reperfusion injury after cerebral ischemia in rats.Methods Sixty adult SD male rats were randomly divided into sham opration group（group A,n =20）,ischemia and reperfusion group（group B,n = 20） and ischemic postprocessing group（group C,n =20）.The model of focal ischemic reperfusion was induced by intraluminal middle cerebral artery occlusion（MCAO） with a nylon monofilament suture.After 24 h of ischemia and reperfusion,the brain tissues were obtained for detecting.Apoptosis of intracephalic nerve cells was detected by in situ end labeling,and contents of Caspase-3,Caspase-9 and cytochrome C were detected by enzyme activity assay,Western blot and immunohistochemisty method.Results The amount of apoptosis cell in group B and C increased when compared with that in group A,and the differences were statistically significant（P 0.05）;the amount of apoptosis cell in group C was significantly reduced when compared with that in group B（P 0.05）.The activity rate of Caspase-3 and Caspase-9 was higher in group B and C than that in group A,while the activity rate in group C was abated when compared to that in group B,and the differences were statistically significant（P 0.05）.Contents of pro-Caspase-3 and pro-Caspase-9 in group B and C were significantly reduced when compared to those in group A（P 0.05）,and the contents in group C were significantly increased when compared to those in group B（P 0.05）.In the cytoplasm,Cyt C was no obviously released in group A,while it was released obviously in group B and C,and the differences were statistically significant（P 0.05）;Cyt C was released more obviously in group C than that in group B（P 0.05）.In mitochondria,Cyt C leakage was more obvious in group C than that in group A,and it was significantly decreased in group C when compared to that in group B,and the differences were statistically significant（P 0.05）

关 键 词：缺血后处理 脑缺血 再灌注损伤 大鼠 CASPASE-3 CASPASE-9 细胞色素C 

分 类 号：R743.3[医药卫生—神经病学与精神病学]