活化的小胶质细胞在大鼠海马神经元缺氧损伤中的作用

目的 探讨缺氧诱导活化的小胶质细胞在SD大鼠海马神经元缺氧损害中的作用机制.方法 建立共培养体系,应用原位缺口末端标记(TUNEL)法、化学发光法探讨不同组别神经元生长状况以及Caspase-3活性;采用免疫荧光法、格里斯试剂法(Griess Reagent)、还原WST-1法、酶联免疫吸咐测定(ELISA)等方法检测各组培养液中NO、O-2以及TNF-α的表达水平.结果 缺氧12h, N9细胞培养液可抑制常规培养的神经元生长增殖活力,同时可加重由缺氧抑制的共培养体系中神经元活力;既可诱导常规培养的神经元凋亡,又可促进共缺氧培养的神经元凋亡;较之于单纯神经元培养系和常规共培养系,共缺氧培养系的培养液中NO、O-2、TNF-α 3类应激性神经毒性因子产量最高.结论 小胶质细胞活化在缺氧诱发的神经元损害中发挥了重要的作用,其活化后产生的神经毒性分子是效应分子.

Action of activated microglia in hippocampal neurons of rat damage induced by hypoxia

Objective In order to investigate the mechanism of hypoxia damage in which the hypoxia-induced- microglia activation acts to hippocampal neurons, the indirect co-culture system was established by using the culture medium which had already trained by microglia cell N9 then used to cultivate fetal rat hippocampal neurons. Methods The co-culture system was established. The neuron growth and Caspase-3 activity of different groups were explored by TUNEL method and chemiluminescence. The expression levels of NO, OSUB>2/SUB>SUP>-/SUP> and TNF-α in different group were detected by immunofluorescence, Griess reagent method, WST-1 method and ELISA method. Results In N9 cells culture medium after 12 hours hypoxia the proliferation, vitality and apoptosis of neuron of conventional cultured and hypoxia cultured were inhibited. The stress-induced neurotoxic factors, NO, OSUB>2/SUB>SUP>-/SUP> and TNF-α in the altogether hypoxic culture system have the highest level than those in the pure culture and conventional co-culture system. Conclusion Microglia activation plays an important role in hypoxia-induced neuronal damage. The effector molecule is the neurotoxic molecule pr