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《Journal of Applied Clinical Pediatrics》 2009-12
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Endoplasmic Reticulum Stress Mediated Neuronal Apoptosis in Hippocampus of Rats with Recurrent Febrile Seizures

CHEN Jing,QIN Jiong,LIU Xiao-yan,HAN Ying,YANG Zhi-xian,CHANG Xing-zhi,JI Xin-na(Department of Pediatrics,Peking University First Hospital,Beijing 100034,China)  
Objective To clarify the involvement of endoplasmic reticulum stress(ERS)response in neuronal apoptosis in the hippocampus of rats with recurrent febrile seizure(FS).Methods Rats aged 21 d(n=80)were randomly divided into 2 groups:control group and FS group,40 rats in each group.In a recurrent FS rat model,the neuronal apoptosis in the hippocampus was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling(TUNEL)method.And the expression patterns of Caspase-12 and glucose-regulated protein 78(GRP78)were detected by Western blot,real-time PCR,and immunohistochemistry.Double fluorescent staining of Caspase-12 and TUNEL method was performed to clarify the neuronal apoptosis involvement of Caspase-12.And double fluorescent staining of Caspase-12 and neuronal nuclei(NeuN)was applied to identify the Caspase-12 positive cells as neurons.Results Apoptotic cells in the hippocampus increased remarkably at 12,24 and 48 h after seizures compared with control rats.In FS group,chromatin condensation and breakdown of the nucleus in neurons in hippocampus were also easily observed by Hoechst staining.Meanwhile,the ERS was induced by recurrent FS,as witness by up-regulated both GRP78 mRNA and protein expression.Western blot revealed that the expression of GRP78 protein in the hippocampus was elevated at 3,6 and 12 h,with the maximum elevation noted at 6 h,after recurrent FS.The GRP78 transcript expression detected by real-time PCR was the same as GRP78 protein.Immunohistochemical analysis showed that GRP78 positive cells were distributed ubiquitously in the hippocampus after recurrent FS.Activated Caspase-12 protein increased at 12,24 and 48 h in FS group.And the Caspase-12 mRNA expression was elevated at 12,24 and 48 h after recurrent FS.Immunoreactivity for Caspase-12 was enhanced in the hippocampus at the same time points,and co-location of Caspase-12 immunoreactivity and DNA fragmentation detected by the TUNEL method.Double fluorescent staining showed that most of Caspase-12 positive cells were neurons.Conclusion The endoplasmic reticulum stress response mediated by Caspase-12 seems to be involved in the neuronal apoptosis caused by recurrent FS.
【Fund】: 国家自然科学基金项目资助(30571969);; 国家985工程重点建设项目资助(985-2-011-24);; 教育部高校博士点基金项目资助(20040001114);; 北京市自然科学基金项目资助(7063096)
【CateGory Index】: R720.597
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