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目的 :研究神经生长因子 (NGF)对过氧化氢 (H2 O2 )诱导神经胶质瘤细胞凋亡的抑制作用及其机制。方法 :抑制细胞增殖的检测采用MTT法 ,吖啶橙 (AO)染色荧光显微观察细胞的形态学变化 ,DNA琼脂糖电泳检测DNA断裂 ,二硫硝基苯甲酸 (DTNB)法检测细胞内还原型谷胱甘肽 (GSH)水平的变化。结果 :10 0～ 80 0μmol/L的H2 O2 明显抑制A 172细胞增殖 ,2 0 0 μmol/LH2 O2 作用 12h后 ,形态学上表现为染色体聚集、核固缩和断裂 ,电泳可见DNA断裂形成的阶梯状条带。经NGF作用 2 4h后可引起A 172细胞内GSH水平上升近 2倍 ,但NGF抑制H2 O2 诱导A 172细胞凋亡的效应与GSH水平无关 ,而且此过程不需新蛋白或RNA的合成。结论 :H2 O2 可有效诱导神经胶质瘤A 172细胞凋亡 ,NGF对此表现显著抑制效应 ,但与细胞内GSH水平变化无关。
Objective: To study the inhibitory effect of nerve growth factor (NGF) on hydrogen peroxide (H2O2)-induced apoptosis of glioma cells and its mechanism. METHODS: MTT assay was used to detect cell proliferation, morphological changes of cells were observed by fluorescence microscopy with acridine orange (AO) staining, DNA fragmentation was detected by DNA agarose gel electrophoresis, and DNA was detected by DTNB method. Prototype changes in glutathione (GSH) levels. RESULTS: The proliferation of A 172 cells was significantly inhibited by H 2 O 2 at 100-80 μmol/L. After 12 h of 200 μmol/L H 2 O 2 , the morphology showed chromosomal aggregation, nuclear condensation, and breakage. DNA fragmentation was observed by electrophoresis. Stepped strips. The effect of NGF on the apoptosis of A 172 cells induced by H2O2 was not related to the level of GSH, and no new protein or RNA synthesis was needed during this process. Conclusion: H2O2 can effectively induce apoptosis of glioma A 172 cells, and NGF has a significant inhibitory effect on this, but it is not related to changes in intracellular GSH levels.