南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (09): 1313-1318.doi: 10.12122/j.issn.1673-4254.2020.09.14

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高丽参热水提取物对神经母细胞瘤细胞parthanatos的影响

郭远波，张登文，龙瑞春，单松贵，孙 强，蔡 彬，王 晟   

  • 出版日期:2020-09-20 发布日期:2020-09-20

Effect of hot water extract of Korean ginseng on neuroblastoma cell parthanatos

  • Online:2020-09-20 Published:2020-09-20

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摘要： 目的 探讨高丽参热水提取物预处理对 N-甲基-N’-硝基-N-亚硝基胍（MNNG）诱导神经母细胞瘤（SH-SY5Y）细胞parthanatos的影响及其机制。方法 将SH-SY5Y细胞分为空白组、MNNG诱导组及高丽参热水提取物预处理组，空白组细胞不给予任何处理，MNNG诱导组给与MNNG 250 μmol/L刺激1 h或4 h，高丽参热水提取物预处理组在给与MNNG诱导前先给与高丽参热水提取物1 mg/mL预处理。以CCK-8和细胞流式技术检测细胞存活率，Western blot检测PAR蛋白，免疫荧光检测 AIF核质分布，细胞流式技术检测细胞活性氧的表达。结果 与空白组比较，SH-SY5Y细胞的存活率随着MNNG刺激的浓度和时间的增加而逐渐降低（P<0.05）；MNNG刺激SH-SY5Y后多聚PAR蛋白表达增加（P<0.05）；高丽参热水提取物预处理可以抑制MNNG诱导的SH-SY5Y死亡，并且降低了MNNG诱导的AIF的表达和入核（P<0.05）；MNNG刺激后活性氧的表达增加，而高丽参热水提取物预处理后，活性氧的表达显著降低（P<0.05）。结论 高丽参热水提取物预处理SH-SY5Y细胞可以降低活性氧的产生并降低了MNNG诱导的parthanatos的发生。

关键词: 高丽参, 热水提取物, parthanatos, 细胞活性氧, 多聚（ADP-核糖）, 凋亡诱导因子

Abstract: Objective To explore the effect of pretreatment of neuroblastoma cells with hot water extract of Korean ginseng on MNNG-induced parthanatos and its mechanism. Methods Neuroblastoma SH-SY5Y cells were pretreated with 1 mg/L hot water extract of Korean ginseng before induction with 250 μmol/L MNNG for 1 h or 4 h. CCK-8 and cell flow cytometry were used to detect cell survival rate. Western blotting was used to detect the changes in poly(ADP-ribose) (PAR) expression in the treated cells. Immunofluorescence assay was used to detect nuclear distribution of apoptosis-inducing factor (AIF), and flow cytometry was used to detect the level of reactive oxygen species (ROS) in the cells. Results Compared with the blank control cells, MNNG-treated SH-SY5Y cells showed significantly decreased survival rate as the concentration of MNNG and the stimulation time increased (P<0.05). Stimulation with MNNG also resulted in significantly increased expression of PAR protein in the cells (P<0.05). Pretreatment of the cells with hot water extract of Korean ginseng obviously inhibited MNNG-induced cell death and significantly reduced AIF expression and nucleation in the cells (P<0.05). MNNG stimulation significantly increased ROS level in the cells, which was decreased significantly by pretreatment of the cells with the extract (P<0.05). Conclusion Pretreatment with hot water extract of Korean ginseng reduces MNNG-induced parthanatos and ROS production in SH-SY5Y cells.

Key words: Korean ginseng, hot water extract, parthanatos, reactive oxygen species, poly(ADP-ribose), apoptosis-inducing factor