摘要
目的探讨缺氧诱导因子-1α（HIF-1α）蛋白及其mRNA在体外培养大鼠皮质神经元缺氧和（或）缺血过程中的表达情况。方法培养16~18 d胎鼠皮质神经元,建立体外神经元缺氧缺血（HI）、单纯缺氧、单纯缺血模型。在缺氧和（或）缺血再灌注后不同时间点,采取免疫细胞化学、原位杂交技术检测皮质神经元HIF-1α蛋白及其mRNA的表达。结果常氧下神经元HIF-1α蛋白有微弱表达,在缺氧和（或）缺血处理后,其表达明显增高,HI再灌注4~8 h后HIF-1α蛋白表达最高,与其他时间点比较均有显著差异（Pa=0）,12 h后开始下降;且HI组HIF-1α蛋白较单纯缺氧和单纯缺血组表达高,差异有统计学意义（Pa=0）。HIF-1αmRNA表达在HI后即达到高峰,2 h后渐下降,在8 h呈低水平表达。结论HIF-1α在缺氧和（或）缺血后的皮质神经元表达的差异具有一定时间规律性,提示对HI神经元进行HIF-1α基因治疗可能是一种可行的方法。
Objective To explore the expression of hypoxia - inducible factor -1α （ HIF -1α） protein and its mRNA in cultured cortical neurons after hypoxia, ischemia or hypoxia - ischemia （HI） and explore the possibilities of HIF -1α gene therapy in HI neurons. Methods The in vitro models of HI,pure hypoxia and pure ischemia were established using embryonic day 16 - 18 rats cortical neurons. and in - situ hybridization were performed to examine the expression of HIF -1α protein and its mRNA at different reperfusion time points in neurons. Results The expression of HIF-1α protein was very week in normoxic cultured neurons, but was up - regulated while treated with hypoxia and（or） ischemia. HIF-1α expression reached peak at 4 to 8 h after reperfusion with HI ,which were statistically significant higher than that at other time points （Pa =0）, and decreased gradually at 12 h. Furthermore, HIF -1α protein expression was significantly higher in HI group compared with that in the pure hypoxia or ischemia group （Pa =0）. HIF -1α mRNA reached peak immediately after HI, decreased gradually at 2 h, and returned to the baseline at 8 h after reperfusion. Conclusions HIF -1α expression on cortical neurons is regulated differently with hypoxia, ischemia or HI treatment, HIF-1α gene therapy for HI neurons maybe a useful method in the future studies.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2007年第8期608-610,共3页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金项目资助(30570623)
四川大学华西医学中心CMB项目资助(00-722)
教育部留学回国人员启动基金项目资助(2006331-11-7)
关键词
缺氧诱导因子-1Α
神经元
缺氧缺血
hypoxia - inducible factor -1α
neurons
hypoxia - ischemia
