缺血缺氧性新生大鼠皮层神经元胞浆Smac／Diablo、caspase-9的表达及参附注射液的干预作用  

Expressions of Smac/Diablo and caspase-9 in the cytoplasm of cerebral cortex of neonatal rats with hypoxic-ischemic brain damage and interference effects of Shenfu injection on them

作　　者：王伟 王军[1] 徐艳[1] 张文辉[1] 周长美[1] 张绍美[1] 

机构地区：[1]徐州医学院附属医院新生儿科,徐州210002 [2]徐州儿童医院新生儿科,徐州210002

出　　处：《中华神经医学杂志》2011年第3期228-231,共4页Chinese Journal of Neuromedicine

基　　金：2008年度省属高校自然科学基础研究计划项目（08KJB320019）

摘　　要：目的 了解缺血缺氧性脑损伤（HIBD）新生大鼠神经元胞浆Smac／Diablo、caspase-9的变化及其相互关系，同时观察参附注射液的十预效果。方法 HIBD模型制作参照Rice等方法。新生7dSD大鼠按照完全随机数字表法分为假手术组、生理盐水对照组与参附治疗组，再按照造模后不同观察时间点又分为3h、6h、12h、24h、3d、7d6个亚组，每亚组8只。参附治疗组于造模后即刻腹腔注射参附注射液10mL／kg，1次／d，连续用药3d。生理盐水对照组用生理盐水替代参附注射液，用法用量相同；假手术组仅切开颈正中皮肤和分离右侧颈总动脉，不结扎，不缺氧。Western blotting检测各时间点各组大鼠神经元胞浆中Smac/Diablo、活化caspase-9蛋白的表达。结果 假手术组Smac/Diablo、caspase-9蛋白表达微弱。生理盐水对照组与参附治疗组2种蛋白造模后3h即明显增加，与假手术组比较，差异有统计学意义（P〈0．05）；24h时2种蛋白表达达到峰值，3d后明显下降，7d时接近假手术组。其中参附治疗组Smac／Diablo蛋白表达量12h、24h、3d、7d时较生理盐水对照组明显下降，差异有统计学意义（P〈0．05）；参附治疗组caspase-9蛋白12h、24h、3d时较生理盐水对照组明显下降，差异有统计学意义（P〈0．05）。结论 （I）HIBD新生大鼠脑皮层神经元存在Smac/Diablo蛋白胞浆转位，同时伴有胞浆活化caspase-9表达增加；参附可减少HIBD大鼠Smac／Diablo蛋白的胞浆转位，减少caspase-9的活化。（2）Smac／Diablo、caspase-9参与了HIBD新生大鼠迟发性神经元损伤的病理机制。Objective To investigate the expressions of Smac/Diablo and caspase-9 and theircorelations in the cytoplasm of cerebral cortex of neonatal rats with hypoxic-ischemic brain damage （HIBD）, and the possible interference effects of Shenfu injection on them. Methods Postnatal 7-d SD rats were randomly divided into Shenfu treatment group （group SF）, normal saline group （group C） and sham-operated group （group S）. Rat models of HIBD were established according to Rice＇s method. Based on the observing times after the success of model making, each group （n=48） was equally divided into 6 subgroups （3, 6, 12 and 24 h, and 3 and 7 d observing groups）. Rats in the group S were not performed ligation of the right common carotid artery and not underwent hypoxia but only performed the separation of right common carotid artery. Shenfu injection was administered by intraperitioneal injection right after H/I insult and then once daily at a dosage of 10 mL/kg for 3 d into the rats of group SF. Saline was also administered into the rats of group C by the same methods and at the same dosage. Cytoplasm proteins were extracted from the cerebral cortex tissue, and then, Western blotting was employed to detect the protein levels of Smac/Diablo and activated caspase-9 in cytoplasm. Results Western blotting showed that the expression levels of Smac/Diablo and activated caspase-9 in cytoplasm of group S were very low; the expression levels of Smac/Diablo and activated caspase-9 in cytoplasm of group SF and group C were increased markedly since the 34 h of the success of model making as compared with those in group S （P〈0.05）： these in the 2 groups reached their peak levels 24 h after H/I insult, gradually decreased 3 d after H/I insult, and almost returned to the original levels 7 d after H/I insult. The expressions of Smac/Diablo in cytoplasm of group SF 12, 24 h and 3 d after H/I insult was significantly lower than that of group C （P〈0.05）; the expression of activated caspase-9 in cytoplasm of group S

关 键 词：细胞凋亡 缺氧缺血性脑损伤 SMAC/DIABLO CASPASE-9 参附注射液 

分 类 号：R722.1[医药卫生—儿科]