大鼠脑出血后蛋白激酶C同工酶和细胞内钙离子与细胞凋亡的关系  

The relationship between protein kinase C isoenzyme,intracellular free calcium and apoptosis after intracerebral hemorrhage in rats

作　　者：李玲[1] 陈康宁[1] 陈福明[1] 黄河清[1] 桂丽[1] 张艳玲[1] 

机构地区：[1]第三军医大学西南医院神经内科,重庆400038

出　　处：《中国脑血管病杂志》2006年第10期440-444,共5页Chinese Journal of Cerebrovascular Diseases

摘　　要：目的 探讨大鼠脑出血后不同时间点血肿周围神经细胞蛋白激酶C（PKC）同工酶（nPKCδ和cPKCγ）、细胞内游离钙浓度与细胞凋亡之间的关系及其意义.方法 将36只成年健康Wistar大鼠,随机分为正常对照组和脑出血后0.5、6、24、72、120h组,每组6只大鼠.采用胶原酶法制备大鼠脑出血模型,利用免疫组化观察PKC同工酶的表达,Fura-2/AM荧光探针法检测细胞内游离钙浓度,TUNEL法检测细胞凋亡.结果 脑出血后血肿周围nPKCδ、cPKCγ表达及细胞内游离钙变化与细胞凋亡数呈正相关,各指标大致为0.5h开始增加,6～24h达高峰,72h开始下降,120 h明显下降（r分别为0.970、0.984,P均＜0.01）.结论 脑出血后血肿周围nPKCδ、cPKCγ表达上调和细胞内钙超载与神经细胞凋亡关系密切,提示细胞信号转导途径异常可能参与了脑出血后神经细胞损伤的病理生理学过程.Objective To explore the relationship between protein kinase C （PKC） isoenzyme （PKCδ, PKCγ） , intracellular free calcium concentration and apoptosis of peri-hematoma neurons at different time points after intracerebral hemorrhage （ ICH ） in rats and their significance. Methods Thirtysix healthy adult Wistar rats were randomized into normal control group and 0. 5, 6, 24, 72, 120 hours groups after ICH （n =6 in each group）. A rat model of ICH was induced by collagenase method. The expression of PKC isoenzyme was observed by immunohistochemistry; the concentration of intracellular free calcium was determined by the Fura-2/AM fluorescent probe, and apoptosis was detected with the TUNEL method. Results The expression of PKCδ, PKCγ and the intracellular free calcium concentration were positively correlated with the numbers of apoptotic neurons around hematomas after ICH （ r = 0. 970, 0. 984. respectively, P 〈 0. 01 ）. They began to increase at 0. 5 hour after ICH, reached their peaks from 6 to 24 hours; and began to decrease at 72 hours, and decreased significantly at 120 hours. Conclusions The expression of PKCδ, PKCγ upregulation and intracellular calcium overload around hematomas after ICH are closely associated with apoptosis. It is suggested that abnormal cell signal transduction may participate in physiopathological processes of neuronal injury after ICH.

关 键 词：脑出血 蛋白激酶C 钙 细胞凋亡 大鼠 

分 类 号：R743.34[医药卫生—神经病学与精神病学]