新生期大鼠反复惊厥后8-OH-dG和CytC的表达与细胞凋亡的关系研究  

The relationship between the expression of 8-OH-dG,CytC and apoptosis in neonatal rats with recurrent seizures

作　　者：刘月影[1] 王春红[1] 崔盈[1] 倪宏[2] 

机构地区：[1]江南大学附属医院儿科,江苏无锡214062 [2]苏州大学附属儿童医院神经病学研究室,江苏苏州215003

出　　处：《重庆医学》2016年第35期4908-4910,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(81471337);无锡市卫计委科研项目(MS201518)

摘　　要：目的探讨新生期大鼠反复惊厥后8-羟基-2′-脱氧鸟嘌呤核苷(8-OH-dG)和细胞色素C(CytC)的表达与细胞凋亡的关系。方法将24只日龄8d的大鼠分为惊厥组和对照组,惊厥组进一步分为惊厥后3h、24h、48h,每组6只。采用吸入三氟乙醚诱导新生期大鼠反复惊厥持续状态模型,酶联免疫吸附试验(ELISA)检测血清8-OH-dG水平,实时荧光定量PCR(RTPCR)和原位末端标记法(TUNEL)分别检测海马CytC的表达和细胞凋亡。结果与对照组相比,TUNEL显示惊厥组海马神经元细胞凋亡明显增多(P<0.01),呈棕黄色颗粒。血清8-OH-dG水平在末次惊厥后各时间点均明显高于对照组(P<0.05或P<0.01)。与对照组相比,CytC在末次惊厥后3h、24h增高(P<0.01),48h后下降至正常水平。8-OH-dG、CytC与细胞凋亡呈正相关(r=0.662、0.565,P<0.05)。结论线粒体损伤可能参与了发育期反复惊厥后脑损伤。Objective To investigate the relationship between the expression of 8-OH-dG,CytC and cell apoptosis in neonatal rats with recurrent seizures. Methods Totally 24 eight-day-old SD rats were randomly divided into two groups： the recurrent seizures group（RS group,30 minutes seizure induced of flurothyl once per day for 7 consecutive days） and control group（CON group, rats were treated without flurothyl）. At designated time points（CON group, 3,24,48 h after the last seizure in RS group） ,the hippocampus of six rats from each group were rapidly dissected out after blood drawn from heart. The 8-OH-dG was detected by ELISA method. The expression of CytC and apoptosis were detected by RT-PCR and TUNEL method respectively. Results Compared with the CON group,TUNEL showed that the apoptotic cell counts in RS group were rapidly increased,especially at 24 h after the last seizures[（54.83 ± 7.16）cells/HPF vs. （15.16 ±2.48）eells/HPF, P〈0.01]. In RS group, the serum levels of 8-OH-dG were significantly higher than that in CON group, particularly at 24 h after the last seizures[（1. 263 83 ± 0. 221 30）pg/μg vs. （0. 822 16±0. 039 380）pg/μg,P〈0.01]. The expression level of CytC mRNA significantly increased at 3 h after the last seizures （0. 114 36±0. 005 28 vs. 0. 093 01±0. 009 79,P〈0. 01） and at 24 h after the last seizures（0. 105 62±0. 007 26 vs. 0. 093 01± 0. 009 79, P〈0.01）, then decreased to normal level. There were obviously positive correlation between 8-OH-dG,CytC and apoptosis（r= 0. 662,0. 565;P〈0.05）. Conclusion The levels of 8-OH-dG and CytC were significantly up-regulated and had positive correlation with apoptosis, suggesting that mitochondrial damage might be involved in the brain injury after the recurrent seizures in neonatal rats.

关 键 词：反复惊厥 8-羟基-2′-脱氧鸟嘌呤核苷 细胞色素C 细胞凋亡 新生大鼠 

分 类 号：R720.597[医药卫生—急诊医学]