CAJ | 학술논문

为了探讨肿瘤坏死因子α(tumor necrosis factor-α,TNFα)刺激的星形胶质细胞对神经元的作用,本研究将体外纯化培养的大鼠海马星形胶质细胞,用反相高效液相法测定TNFα刺激后细胞培养液内谷氨酸的含量;将TNFα刺激后的星形胶质细胞条件培养液(astrocytic conditioned medium,ACM)作用于培养的海马神经元,运用免疫细胞化学方法和图像分析技术研究神经元NF κBp65和谷氨酸的表达.结果表明:(1)TNFα可明显促进星形胶质细胞释放谷氨酸,(2)ACM作用15 min即可诱导神经元NFκBp65的核表达,30 min达高峰,180 min恢复至对照水平,(3)ACM作用60 min可使谷氨酸免疫反应阳性神经元平均光密度明显升高,持续至240 min.提示,TNFα刺激的星形胶质细胞可通过释放谷氨酸等可溶性物质使神经元快速激活、兴奋性升高.
위료탐토종류배사인자α(tumor necrosis factor-α,TNFα)자격적성형효질세포대신경원적작용,본연구장체외순화배양적대서해마성형효질세포,용반상고효액상법측정TNFα자격후세포배양액내곡안산적함량;장TNFα자격후적성형효질세포조건배양액(astrocytic conditioned medium,ACM)작용우배양적해마신경원,운용면역세포화학방법화도상분석기술연구신경원NF κBp65화곡안산적표체.결과표명:(1)TNFα가명현촉진성형효질세포석방곡안산,(2)ACM작용15 min즉가유도신경원NFκBp65적핵표체,30 min체고봉,180 min회복지대조수평,(3)ACM작용60 min가사곡안산면역반응양성신경원평균광밀도명현승고,지속지240 min.제시,TNFα자격적성형효질세포가통과석방곡안산등가용성물질사신경원쾌속격활、흥강성승고.
To observe the effect of astrocytes activated by stimulation of tumor necrosis factor-α(FNFα) on neurons, the contentof glutamate in the supernatant of purified cultured hippocampal astrocytes stimulated by TNFα(1ng/ml) of rat were measuredby high performance liquid chromatography(HPLC). The astrocytic conditioned medium(ACM) stimulated by TNFα was thencollected and added to the supernatant of cultured hippocampal neurons. The expression of NF-κBp65 and glutamate was studiedby immunocytochemical method. The results showed that (1) TNFα could promote the release of glutamate from astrocytes,(2) The expression of NF-κBp65 in the nucleus of neurons began to be induced at 15 min after ACM. The percentage and averageoptical density of the NF-κBp65- immunoreactivity(IR) positive nucleus reached the maximal level at 30 min and returned to thecontrol level by 180 min, (3) The average optical density of Glu-IR positive neurons began to increase markedly 60 min afterACM and refuned to the normal at 240 min. The results indicate that neurons can be quickly activated and the excitability of neu-rons can also be elevated through the neuro active substance such as glutamate released by astrocytes pretreated with TNFα.