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目的研究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 4-甲基偶氮唑蓝(MTT)检测PC12细胞活性;Hochest 33258染色观察细胞凋亡的变化;DCFH-DA/DAF-FM DA检测ROS/NO表达情况;Westernblotting检测Nrf2、Keap1、SOD1、SOD2、CAT蛋白的表达情况。结果 MPP+(500μmol·L~(-1))作用于PC12细胞24 h后,与正常组比较,细胞存活率降低至(51.3±2.2)%(P<0.01);TSG(10,50μmol·L~(-1))预处理24 h后,细胞存活率增加至(60.1±1.5)%,(74.2±2.1)%,(82.1±1.5)%(P<0.05);细胞核固缩明显减少,且具有剂量依赖性关系。此外,MPP+处理后使PC12细胞中ROS/NO升高,Nrf2、SOD1、SOD2、CAT蛋白表达减少,使keap1表达增多,而TSG预处理可以明显改善这些变化。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,其作用机制可能通过激活Nrf2/Keap1通路减轻细胞内活性氧的聚集有关。
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 The apoptosis of PC12 cells was detected by methyl thiazolyl tetrazolium (MTT) assay. The changes of apoptosis were observed by Hochest 33258 staining. The expression of ROS / NO was detected by DCFH-DA / DAF-FM DA. The levels of Nrf2, SOD2, CAT protein expression. Results After treated with MPP + (500μmol·L -1) for 24 h, the survival rate of PC12 cells was decreased to (51.3 ± 2.2)% compared with that of normal cells (P <0.01). TSG (10,50μmol·L -1) (P <0.05). The cell nuclear shrinkage was significantly decreased after pretreatment for 24 h (60.1 ± 1.5)%, (74.2 ± 2.1)% and (82.1 ± 1.5)% Dose-dependent relationship. In addition, MPP + treatment increased PC12 cells ROS / NO, Nrf2, SOD1, SOD2, CAT protein expression decreased, keap1 expression increased, and TSG pretreatment can significantly improve these changes. Conclusion TSG can inhibit the apoptosis of PC12 cells induced by MPP + in a concentration-dependent manner. The mechanism may be related to the reduction of intracellular reactive oxygen species accumulation by activating Nrf2 / Keap1 pathway.