高丽参热水提取物对神经母细胞瘤细胞parthanatos的影响  

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

作　　者：郭远波[1] 张登文[1] 龙瑞春[1] 单松贵 孙强[1] 蔡彬 王晟[1] 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)

机构地区：[1]广东省人民医院//广东省医学科学院麻醉科,广东广州510080 [2]广东省人民医院//广东省医学科学院科教处,广东广州510080

出　　处：《南方医科大学学报》2020年第9期1313-1318,共6页Journal of Southern Medical University

基　　金：广东省中医药局科研项目(20191006);广东省医学科研基金(A2020038);广东省基础与应用基础研究基金项目(2019A1515110063)。

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 250μmol/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。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 250μmol/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.

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 

分 类 号：R285.5[医药卫生—中药学]