摘要
目的观察惊厥持续状态(SC)幼年大鼠海马前凋亡因子CHOP动态表达及神经细胞凋亡的变化,探讨依达拉奉(ED)对二者的影响。方法将195只SD幼年雄性大鼠随机分为9 g.L-1盐水对照组(NS组)、SC组和ED组,每组65只,各组均按SC后处死时间点分为2 h、12 h、24 h、48 h、72 h 5个亚组,每组13只。应用氯化锂-毛果芸香碱建立大鼠SC模型,用半定量反转录-PCR(RT-PCR)动态观察SC后大鼠海马CHOP mRNA的表达,采用免疫组织化学SABC法检测海马CA1区CHOP蛋白表达;并用HE染色观察海马CA1区病理改变,原位细胞凋亡检测法(TUNEL)观察海马CA1区神经元凋亡细胞数。结果 1.RT-PCR法:SC组幼年大鼠海马CHOP mRNA表达于2 h开始增加,于12 h达高峰,之后开始下降;与NS组各时间点比较差异均有统计学意义(Pa<0.01);与SC组比较,ED组(2 h,12 h,48 h)CHOP mRNA表达均明显下降(Pa<0.05,0.01)。2.免疫组织化学结果:SC组幼年大鼠海马CHOP蛋白表达于2 h开始增加,12 h明显增加,24 h达高峰;与NS组各时间点比较差异均有统计学意义(Pa<0.01);ED组各时间点均较SC组明显降低(Pa<0.01,0.05)。3.TUNEL结果:SC组海马CA1区TUNEL阳性细胞数于惊厥后24 h迅速增加,48 h达高峰(Pa<0.05,0.01);与NS组比较,12～72 h时间点组均明显高于NS组(Pa<0.01);与SC组比较,ED组12～72 h CA1区TUNEL阳性细胞数明显减少(P<0.05,0.01)。4.HE染色:SC后出现神经元变性及丢失,48 h组病理改变最显著,与TUNEL表达一致;而ED组病理改变减轻。结论 SC后早期幼年大鼠可能触发了内质网应激中CHOP介导的凋亡信号途径,从而引起了脑损伤;而ED可能通过下调其表达,从而缓解惊厥后脑损伤。
Objective To investigate the dynamic expressions of CHOP in juvenile rat hippocampus after status convulsion,and the changes in apoptosis of nerve cells,and to explore the effects of edaravone（ED） on them. Methods One hundred and ninety-five juvenile male Sprague-Dawley（SD）rats were randomly divided into 3 groups.Each group were further divided into 5 subgroups in different executed time points（2 h,12 h,24 h,48 h,72 h）,including 9 g·L-1 saline control group（NS group）,status epilepticus group（SC group）and ED treatment group（ED group） after SC.The rats in SC group were kindled into epilepsy by lithium-pilocarpine chemical method.The expression of CHOP mRNA was detected with reverse transcription-polymerase chain reaction（RT-PCR）method.Expression of CHOP protein was detected with immunohistochemistry methods.The neuron apoptosis was observed by TdT-mediated dUTP nick end labeling. Results 1.After RT-PCR assay,the expression of CHOP mRNA in the hippocampus in SC group began increasing at 2 h,and reached a maximum at 12 h,and then began decreasing at 24 h.There were significant differences between SC group and NS group（Pa0.01）.And the expression of CHOP mRNA in ED group was much lower than that in SC group（Pa0.05,0.01）.2.Measured by immunohistochemistry,the value of optical density of CHOP positive cells began increasing in SC group at 2 h,and reached a maximum at 12 h,and decreased remarkably at 24 h.There was significant difference compared with NS group（Pa0.01）.And the value of optical density of CHOP positive cells in ED group was much lower than that in SC group（Pa0.01,0.05）.3.The TUNEL positive cells in hippocampus CA1 of SC group were more than those in NS group after the SC 12 h（Pa0.01）,TUNEL positive cells showed a significant drop in SC group at 12-48 h time points,and reached its highest level at 48 h（P0.01,0.05）.There were different degrees degeneration and cell loss in hippocampus neurons in SC group,and the changes were similar with the TUNEL. Conclusi
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第18期1432-1435,1438,共5页
Journal of Applied Clinical Pediatrics
关键词
脑损伤
内质网应激
原位细胞凋亡检测法
惊厥
依达拉奉
brain injury
endoplasmic reticulum stress
TdT-mediated dUTP nick end labeling
convulsion
edaravone
