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目的:观察6-羟基多巴(6-OHDA)诱导PC12细胞凋亡的作用,探讨神经生长因子(NGF)对PC12细胞凋亡的保护作用。方法:将6孔培养板中的PC12细胞分别加入不同浓度的6-OHDA(0,25,50,100,150和200μmol/L);同样6-OHDA各组,每孔均加入终浓度为100μg/L的NGF。分别干预24h后终止培养,观察PC12细胞凋亡的程度。用Hoechst33258细胞核染色法、凋亡细胞的DNA裂解分析—琼脂糖凝胶电泳、TUNEL原位凋亡检测和计算机自动图像分析等方法进行凋亡检测。结果:6-OHDA50,100,150和200μmol/L均不同程度地诱导了PC12细胞凋亡,表现剂量效应。NGF对6-OHDA诱导的PC12细胞凋亡具有保护作用犤NGF组比8KNG组:细胞核面积:(56.48±3.06)比(36.19±2.15)μm2,光密度:0.38±0.03比0.53±0.3,t=2.36,3.07,P<0.05。结论:支持6-OHDA在体外能够诱导PC12细胞凋亡的观点,并且有剂量效应。NGF对6-OHDA诱导的PC12细胞凋亡具有保护作用。
Objective: To observe the effect of 6-OHDA on the apoptosis of PC12 cells and to explore the protective effect of nerve growth factor (NGF) on the apoptosis of PC12 cells. Methods: PC12 cells in 6-well plates were added with different concentrations of 6-OHDA (0, 25, 50, 100, 150 and 200 μmol / L) . After 24h intervention, the cells were killed to observe the degree of PC12 cell apoptosis. Apoptosis was detected by Hoechst33258 nuclear staining, DNA lysis assay of apoptotic cells - agarose gel electrophoresis, TUNEL in situ apoptosis assay and computer automated image analysis. Results: 6-OHDA at doses of 100, 100, 150 and 200 μmol / L induced apoptosis of PC12 cells to varying degrees and showed dose-response. NGF had a protective effect on 6-OHDA-induced apoptosis in PC12 cells. Compared with 8KNG group, NGF group had a nucleus area of (56.48 ± 3.06) vs (36.19 ± 2.15) μm 2, optical density of 0.38 ± 0.03 vs 0.53 ± 0.3, t = 2.36, 3.07, P <0.05. CONCLUSION: The support of 6-OHDA in vitro can induce apoptosis in PC12 cells with a dose-response. NGF has a protective effect on 6-OHDA-induced PC12 cell apoptosis.