目的:研究两种体外温育的晚期糖基化终产物(AGE)是否可以诱导星型胶质细胞分泌白介素-1β和肿瘤坏死因子α,并导致氧应激增加、一氧化氮释放.方法:RT-PCR技术检测两种细胞因子水平及AGE受体(RAGE)存在与否;DTNB反应测量还原型谷胱甘肽的水平:利用Griess试剂测量一氧化氮含量.结果:AGE 1 g/L(尤其是半乳糖温育产物)作用72小时后使星型胶质细胞培养上清和细胞裂解物的细胞因子含量显著升高.且呈剂量依赖性.半定量RT-PCR证明两种细胞因子的变化是由于其转录水平增加所致.AGE还可导致星型胶质细胞内还原性谷胱甘肽减少,一氧化氮释放.RAGE存在于此类星型胶质细胞中.结论:AGE可诱导星型胶质细胞分泌炎性因子白介素-1β和肿瘤坏死因子α,并升高氧应激水平,这至少部分解释了AGE在神经变性性疾病如阿尔采默病和帕金森氏病以及脑衰老中的负性作用.%AIM: To investigate whether two kinds of in vitro prepared advanced glycation end products (AGE), Glu-BSA andGal-BSA, could induce proinflammatory mediators IL-1β and TNF-α, as well as oxidative stress and nitric oxide(NO), in astrocytes, thus contributing to brain injury. METHODS: Radioimmunoassay and RT-PCR techniquewere used to detect two cytokines' level and existence of receptor for AGE (RAGE). DTNB reaction was used tomeasure reduced glutathione (GSH) level. NO content was assayed using Griess reagent provided by Promega.RESULTS: Enhanced protein levels of both cytokines in supernatants and cell lysates of astroglia cultures weredetected after treated with AGE-BSA 1 g/L, especially Gal-BSA, for 72 h. The increases were also in a concentra-tion-dependent manner. Changes in protein levels might be attributed to changes in transcriptional levels docu-mented by semi-quantitative RT-PCR. Both AGE-BSA could also reduce astrocytic GSH and induce NO release.RAGE was detected in astrocytes. CONCLUSION: Enhanced levels of astrocytic proinflammatory mediators IL-1 5 and TNF-α, and oxidative stress caused by AGE might contribute to, at least partially, the detrimental effects ofAGE in neuronal disorders and aging brain.
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