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目的:探讨2,3,5,4’-四羟基二苯乙烯-2-o-β-D-葡萄糖苷(2,3,5,4’-tetrahydroxystibene-2-o-β-D-glucoside,TSG)对1-甲基-4-苯基吡啶离子(1-methy-4-phenylpyridinium,MPP+)诱导PC12细胞凋亡的影响及其可能机制。方法:四甲基偶氮唑蓝(MTT)比色试验检测PC12细胞活性;Hoechst33258染色法测定细胞凋亡;Westernblotting检测NF-κB(P65)和IκBα蛋白的表达。结果:MPP+(300μmol/L)作用于PC12细胞24h后,与正常对照组比较,细胞存活率降低(53.3±3.4%)(P<0.01);细胞染色质固缩,细胞核呈致密浓染。TSG(1,5,10μmol/L)预处理24h后,细胞存活率增加(60.8±1.9%),(70.1±1.8%)(P<0.01),(81.2±1.9%)(P<0.01);细胞核凝聚明显减少,且具有量一效关系。另外,MPP+可使PC12细胞核中NF-κB(P65)蛋白表达升高,细胞浆中IκBα蛋白表达降低;与MPP+处理组细胞相比,TSG预处理后,PC12细胞核中高表达的NF-κB(P65)蛋白水平明显降低,细胞浆中低表达的IκBα蛋白水平升高。结论:TSG对MPP+诱导的PC12细胞凋亡具有浓度依赖性的抑制作用,其作用机制可能与抑制NF-κB的激活有关。
Objective: To investigate the effects of 2,3,5,4’-tetrahydroxystibene-2-o-β-D-glucoside TSG) on the apoptosis of PC12 cells induced by 1-methyl-4-phenylpyridinium (MPP +) and its possible mechanism. Methods: MTT assay was used to detect the activity of PC12 cells. Hoechst33258 staining was used to detect apoptosis. Western blotting was used to detect the expression of NF-κB (P65) and IκBα protein. RESULTS: After treated with MPP + (300μmol / L) for 24 hours, the cell viability decreased (53.3 ± 3.4%) compared with the normal control group (P <0.01). The chromatin was condensed and the nucleus was densely stained. Cell viability increased by 60.8 ± 1.9% (70.1 ± 1.8%) (P <0.01), (81.2 ± 1.9%) (P <0.01) after pretreated with TSG (1,5,10 μmol / L) Significantly reduce the nuclear condensation, and the relationship between quantity and effectiveness. In addition, MPP + increased the expression of NF-κB (P65) in PC12 cells and decreased the expression of IκBα in cytoplasm. Compared with MPP + -treated cells, the expression of NF-κB (P65) ) Protein levels were significantly reduced, low expression of cytoplasmic IκBα protein levels. CONCLUSION: TSG can inhibit the apoptosis of PC12 cells induced by MPP + in a concentration-dependent manner, which may be related to the inhibition of the activation of NF-κB.