HIF-1α在缺糖缺氧诱导的原代皮质神经元necroptosis中的表达  

HIF - 1α expression in oxygen - glucose deprivation - induced primary cortical neuron necroptosis

作　　者：张翠翠[1] 陈巍巍[1] 徐兴顺[2] 耿德勤[3] 

机构地区：[1]徐州医学院2008级研究生,江苏徐州221002 [2]苏州大学第二附属医院神经内科,江苏苏州215004 [3]徐州医学院附属医院神经内科,江苏徐州221002

出　　处：《徐州医学院学报》2011年第5期293-296,共4页Acta Academiae Medicinae Xuzhou

基　　金：基金项目：国家自然科学基金（30700245）

摘　　要：目的 探讨低氧诱导因子-1α（hypoxia-inducible factor-1α,HIF-1α）是否参与缺糖缺氧诱导的原代皮质神经元necroptosis.方法 原代皮质神经元培养14天,予caspase抑制剂Z-VAD-FMK（20 μmol/L）预保护30 min,缺糖缺氧2 h,再灌注0、2、6、12、24、48 h,进行LDH测定;RT-PCR测HIF-1α RNA表达;Western blot检测总HIF-1α蛋白表达情况;分别提取细胞质和细胞核蛋白,Western blot分别检测胞质、胞核内HIF-1α蛋白表达情况.结果 caspase抑制剂Z-VAD-FMK预作用30 min、缺糖缺氧2 h、再灌注2 h后培养基中LDH增加（P＜0.05）,再灌注12 h达高峰;缺糖缺氧后HIF-1α RNA表达无变化（P＞0.05）;HIF-1α总蛋白表达增加（P＜0.05）,再灌注12 h达高峰;再灌注6 h时细胞质HIF-1α蛋白表达达高峰（P＜0.05）,随后降低;再灌注12 h时,细胞核HIF-1α蛋白表达达高峰（P＜0.05）,随后降低.结论 HIF-1α参与缺糖缺氧诱导的原代皮质神经元necroptosis.Objective To investigate whether hypoxia - inducible factor - 1 α （ HIF - 1α） is involved in primary cortical neuronal necroptosis induced by oxygen - glucose deprivation （OGD）. Methods Primary cerebrocortical neurons were cultured for 14 days, followed by pretreatment with Z -VAD - FMK （20μmol/L） for 30 min prior to exposure to OGD for 2 hours （h） and reoxygenation for different durations （0, 2, 6, 12, 24 and 48 hours）. Cell viability was determined by determination of LDH level; RTPCR was employed to determine the expression of HIF- 1 α RNA, and the expression of total proteins of HIF - 1αwas determined by Western blot analysis; the cytoplasmic and nuclear proteins of HIF - 1α were respectively extracted for the determination of cytoplasmic and nuclear protein expression levels by Western blot. Results There was an increase in LDH level in the culture medium following 30 - minute pretreatment with caspase inhibitor Z - VAD - FMK, exposure to OGD for 2 h and reperfusion for 2 h （P 〈0.05）, with its peak at 12 h of reperfusion; there was no change in the expression of HIF - 1 α RNA subsequent to OGD （ P 〉 0.05） ; the expression of total proteins of HIF- 1α was upregulated （P 〈0.05）, with its peak at 12 h of reperfusion; the protein expression of cytoplasmic HIF - 1 α reached its peak at 6 h of reperfusion （P 〈 O. 05 ） and declined thereafter; the protein expression of nuclear HIF - 1 α reached its peak at 12 h of reperfusion （ P 〈 0.05 ） and declined afterwards. Conclution HIF - 1α was involved in oxygen -glucose deprivation- induced primary cortical neuronal necroptosis.

关 键 词：低氧诱导因子-1Α NECROPTOSIS 缺糖缺氧 原代皮质神经元 

分 类 号：R743[医药卫生—神经病学与精神病学]