参附注射液对缺氧缺血性脑损伤新生大鼠脑皮层IRE1α表达及神经元凋亡的影响  

Effect of Shenfu injection on IRE1α expression in cerebral cortex of neonatal rats with hypoxic-ischemic brain damage

作　　者：刘文强[1] 王军[1] 杨倩倩[1] 徐艳[1] 

机构地区：[1]徐州医学院附属医院新生儿科,徐州221002

出　　处：《神经解剖学杂志》2013年第6期671-676,共6页Chinese Journal of Neuroanatomy

基　　金：江苏省省属高校自然科学基础研究计划项目(08KJB320019)

摘　　要：目的:观察参附注射液对缺氧缺血性脑损伤(HIBD)新生大鼠脑皮层中内质网应激关键分子-肌醇需求因子1α(inositol-requiring enzyme1α,IRE1α)表达及神经元凋亡的影响,探讨其脑保护机制。方法:参照Rice法制备HIBD模型,将7日龄SD大鼠随机分为假手术组(S)和HIBD模型组,模型组又分为生理盐水对照组(C)和参附治疗组(SF),各组按照观察时间点不同进一步分为3、6、12、24、72 h 5个亚组,每组均10只。HE染色光镜下观察脑组织病理变化;RT-PCR方法检测IRE1αmRNA的表达变化;Western blot方法检测IRE1α蛋白表达变化;流式细胞术检测神经元的凋亡率;结果:(1)RT-PCR:C组在HIBD后3、6、12、24 h时间点IRE1αmRNA表达量较S组升高(P<0.05);SF组各时间点IRE1αmRNA表达均较S组升高(P<0.05)。SF组在6、12、24、72 h时间点的IRE1αmRNA表达量较C组升高明显(P<0.05)。(2)Western blot:C组和SF组各时间点IRE1α蛋白表达量均较S组升高(P<0.05),SF组在6、12、24、72 h时间点的IRE1α蛋白表达量较C组升高明显(P<0.05)。(3)神经元的凋亡率:SF组各时间点的凋亡率均明显低于C组(P<0.05)。结论:参附注射液可能通过上调IER1α的表达,减少神经元凋亡来减轻新生大鼠缺氧缺血性脑损伤。Objective： To investigate the expression of the key marker of endoplasmic reticulum stress- IRE1α （inositol- requiring enzyme 1α） and neuronal apoptosis in cerebral cortex of neonatal rats with hypoxic-ischemic brain damage （HIBD）, and the neuroprotective role of Shenfu injection. Methods： Sprague-Dawley （SD） rats aged 7 days were ran- domly assigned to three groups： sham operation group （S group）, saline control group （C group） and Shenfu injection treatment group （SF group）. The rat HIBD models were established according to Rice＇s method. Based on the observing time points, each group was subdivided into 5 groups （n = 10）, sacrificed at 3,6,12,24 and 72 h. The pathological chan- ges of brain tissue were observed under light microscope. The expression of IRElct mRNA in the cerebral cortex was de- tected by RT-PCR. The change of IREla protein in the cerebral cortex was detected by Western blot. The apoptosis rate was measured by flow cytometry. Results： （1）RT-PCR：Compared with the S group, the expression of IRElot mRNA in the C group was obviously upregulated at 3,6,12 and 24 h after HIBD （P 〈 0.05 ）, the expression of IRElct mRNA inthe SF group was obviously upregulated at 3, 6,12,24 and 72 h after HIBD （P 〈0.05）. The expression of IRE1α mR- NA in SF group was notably higher than the C group at 6,12,24 and 72 h after HIBD （ P 〈 O. 05 ）. （2） Western blot ： the expression of IRE1αprotein in the C group and SF group was obviously upregulated compared with the S group （P 〈 O. 05）. IRElct protein expression in the SF group was notably higher than the C group at 6,12,24 and 72 h after HIBD （P 〈0.05 ）. （3）The apoptosis of neurons in cerebral cortex in every time point in SF group was significantly less than that of C group （ P 〈 0.05 ）. Conclusions： Shenfu injection may have neuroprotection against HIBD by up-regulation of IRE1α expression.

关 键 词：参附注射液 缺氧缺血性脑损伤 肌醇需求因子1α 内质网应激 凋亡 大鼠 

分 类 号：R651.1[医药卫生—外科学]