缺氧缺血性脑损伤新生大鼠XIAP、Caspase-12的表达及参附注射液对其影响  

Expression of XIAP,Caspase-12 in Neonatal Rats with Hypoxic-Ischemic Brain Damage and Protective Mechanism of Shenfu Injection

作　　者：吴宏伟 王军[2] 李振光 王伟 刘金凤 

机构地区：[1]徐州市儿童医院新生儿科,江苏徐州221006 [2]徐州医学院附属医院新生儿科,江苏徐州221002

出　　处：《药物生物技术》2014年第4期334-337,共4页Pharmaceutical Biotechnology

基　　金：徐州市科技指令性计划项目(No.XF10C033)

摘　　要：研究新生大鼠脑缺氧缺血后X-染色体连锁的细胞凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、半胱氨酸天冬氨酸特异性蛋白酶-12(Caspase-12)在脑海马神经元细胞中的动态变化及参附注射液对其影响。新生7 d SD大鼠随机分为3组:假手术组(S组)、脑缺氧缺血(HI)+生理盐水对照组(C组)、HI+参附治疗组(SF组),每一组按照观测的时间点不同分为3 h,6 h,12 h,24 h,3 d,7 d 6个亚组,每组8只,用Rice法制备新生大鼠缺氧缺血性脑损伤(HIBD)模型,在每个时间点将大鼠断头取右侧海马脑组织匀浆,用RT-PCR的方法检测脑海马组织Caspase-12mRNA、XIAPmRNA的表达,用TUNEL染色法检测相应时间点光镜下脑海马组织的凋亡。脑缺氧缺血后3 h XIAP mRNA表达明显上调并达到峰值,持续高表达至3 d(P<0.01或0.05),7 d与假手术组比较无统计学意义(P>0.05);参附组XIAP mRNA表达在3,6,12,24 h较生理盐水对照组比较明显上调(P<0.01),在缺血后3 d与对照组之间差异无统计学意义。S组右侧海马神经元Caspase-12mRNA表达很弱;C组Caspase-12mRNA表达在12 h达到峰值,SF组Caspase-12mRNA表达在6 h,12 h,24 h,3 d较C组明显下降(P<0.001)。TUNEL显示HI后3hC组和SF组即发现结扎侧脑海马有少量的凋亡细胞,随着缺氧缺血时间的延长,凋亡细胞逐渐增多,24 h达到高峰后下降;SF组在HI后24 h,3 d,7 d均明显低于C组(P<0.05)。新生大鼠缺氧缺血后脑损伤后XIAP、Caspase-12表达均是增加的,但各自有其表达时相特异性,HI后脑组织细胞的凋亡明显增加,Caspase-12、XIAP参与了新生大鼠HIBD后神经元损伤的病理机制;参附注射液能够上调XIAP、下降Caspase-12的表达,从而起到对HIBD新生大鼠保护作用。To investigate the expression of XIAP and Caspase-12 in the neuronal cytoplasm in cerebral hippocampus of neonatal rats with hypoxic-ischemic brain damage（ HIBD） and to explore the effect of Shenfu injection on it,neonatal SD rats were randomly divided into three equal groups： 1neonatal rats underwent sham operation but did not undergo hypoxia as control group（ Group S）;2normal saline（ NS） group（ Group C）; 3Shenfu group（ Group SF）. They were further divided into six subgroups（ n =8 in each subgroup） according to the time points at 3 h,6 h,12 h,24 h,3 d and 7 d. After HI insult rats from each group were sacrificed with their right cerebral hippocampine tissue taken out. XIAP mRNA and Caspase-12 mRNA expression in cerebral hippocampus tissue were assessed by RT-PCR. The apoptosis cells in hippocampus were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling（ TUNEL）. The expression of XIAP mRNA in Group SF and Group C was clear increased and reached peak at 3 h after HI insult,then overexpressed to 3 d and almost returned to the original levels 7 d after HI and had no significant differences compared with Group S. The expression 3 h,6 h,12 h,24 h of XIAPmRNA in group SF was markedly higher than that of group C（ P 0. 05 or 0. 01）. The expression of XIAP 3 d after HI insult in group SF compared with group C had no difference（ P 0.05）. The expression of Caspase-12 mRNA in Group S was very low. RT-PCR showed Caspase-12 mRNA in Group C reached a peak at 12 h post-HI. The expression of Caspase-12 mRNA in Group SF at 6 h,12 h,24 h and 3 d were significantly lower than those in Group C（ P 0. 01 and 0. 05）. TUNEL staining showed that the number of apoptotic cells in Group SF and Group C in the cerebral hippocampus of ipsilateral hemisphere was gradually increased at 3 h and peaked at 24 h,and then decreased after HI insult. The number of apoptotic cells in Group SF at 24 h,3 d and 7 d was significantly lower than those in Group C（ P 0. 05 and0. 0

关 键 词：缺氧缺血性脑损伤 X-染色体连锁细胞凋亡抑制蛋白 半胱氨酸天冬氨酸特异性蛋白酶-12 参附注射液 细胞凋亡 

分 类 号：R965[医药卫生—药理学]