Guo Yuanbo, Zhang Dengwen, Long Ruichun, Shan Songgui, Sun Qiang, Cai Bin, Wang Sheng
Department of Anesthesiology, Guangdong Provincial People's Hospital/Guangdong Academy of Medical Sciences, Guangzhou 510080, China.
Department of Science and Education, Guangdong Provincial People's Hospital/Guangdong Academy of Medical Sciences, Guangzhou 510080, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Sep 30;40(9):1313-1318. doi: 10.12122/j.issn.1673-4254.2020.09.14.
To explore the effect of pretreatment of neuroblastoma cells with hot water extract of Korean ginseng on MNNG-induced parthanatos and its mechanism.
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.
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 ( < 0.05). Stimulation with MNNG also resulted in significantly increased expression of PAR protein in the cells ( < 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 ( < 0.05). MNNG stimulation significantly increased ROS level in the cells, which was decreased significantly by pretreatment of the cells with the extract ( < 0.05).
Pretreatment with hot water extract of Korean ginseng reduces MNNG-induced parthanatos and ROS production in SH-SY5Y cells.
探讨人参热水提取物预处理神经母细胞瘤细胞对N-甲基-N'-硝基-N-亚硝基胍（MNNG）诱导的副凋亡的影响及其机制。
用1mg/L人参热水提取物预处理神经母细胞瘤SH-SY5Y细胞，然后用250μmol/L MNNG诱导1小时或4小时。采用细胞计数试剂盒-8（CCK-8）法和细胞流式术检测细胞存活率。采用蛋白质免疫印迹法检测处理后细胞中聚（ADP-核糖）（PAR）表达的变化。采用免疫荧光分析法检测凋亡诱导因子（AIF）的核分布，采用流式细胞术检测细胞内活性氧（ROS）水平。
与空白对照细胞相比，随着MNNG浓度增加和刺激时间延长，MNNG处理的SH-SY5Y细胞存活率显著降低（P<0.05）。MNNG刺激还导致细胞中PAR蛋白表达显著增加（P<0.05）。用人参热水提取物预处理细胞明显抑制了MNNG诱导的细胞死亡，并显著降低了细胞中AIF的表达和成核（P<0.05）。MNNG刺激显著增加细胞内ROS水平，而提取物预处理细胞可显著降低该水平（P<0.05）。
人参热水提取物预处理可降低MNNG诱导的SH-SY5Y细胞副凋亡及ROS生成。
