食品科学 ›› 2020, Vol. 41 ›› Issue (15): 194-200.doi: 10.7506/spkx1002-6630-20190714-185

• 营养卫生 • 上一篇    下一篇

枸杞多糖联合顺铂对人肺腺癌细胞A549氧化损伤及凋亡的影响

韩何丹,韩照玉,杜月梅,刘云帆,李艾琳,高丽萍   

  1. (北京联合大学生物化学工程学院,生物活性物质与功能食品北京市重点实验室,北京 100191)
  • 出版日期:2020-08-15 发布日期:2020-08-19
  • 基金资助:
    北京市自然科学基金项目(7163211)

Effects of Lycium barbarum Fruit Polysaccharides Combined with Cisplatin on Oxidative Damage and Apoptosis in Human Lung Adenocarcinoma Cell Line A549

HAN Hedan, HAN Zhaoyu, DU Yuemei, LIU Yunfan, LI Ailin, GAO Liping   

  1. (Beijing Municipal Key Laboratory of Biologically Active Substances and Functional Food, College of Biochemical Engineering, Beijing Union University, Beijing 100191, China)
  • Online:2020-08-15 Published:2020-08-19

摘要: 目的:探讨枸杞多糖(Lycium barbarum polysaccharides,LBP)联合顺铂(cisplatin,DDP)对人肺腺癌细胞A549氧化损伤、细胞凋亡及凋亡相关蛋白表达的影响。方法:细胞分为对照组、DDP组(6 mg/L)、LBP组(8 mg/L)和DDP(6 mg/L)+LBP(8 mg/L)组,CCK-8法检测DDP、LBP单独及联合用药对A549细胞存活率的影响;黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(superoxide dismutase,SOD)活力;二硫代二硝基苯甲酸法测定谷胱甘肽(glutathione,GSH)含量;硫代巴比妥酸法测定丙二醛(malondialdehyde,MDA)含量;流式细胞术检测活性氧(reaction oxygen species,ROS)含量和细胞凋亡率;Western blot检测细胞凋亡相关蛋白Bcl-2、Bax和Caspase-3的表达。结果:LBP能极显著增强DDP对A549细胞存活率的抑制作用(P<0.01)。DDP导致A549细胞SOD活力和GSH含量极显著降低(P<0.01),MDA含量和ROS含量极显著升高(P<0.01)。而LBP联合DDP组与DDP组比较,细胞内SOD活力、GSH含量、MDA含量无显著变化(P>0.05),而ROS含量明显降低(P<0.01)。一定质量浓度DDP和LBP单独及联合处理均能极显著促进细胞凋亡(P<0.01),并且能极显著降低Bcl-2、提高Bax、Caspase-3表达量及Bax/Bcl-2值(P<0.01)。结论:LBP与DDP联合作用可明显促进A549细胞凋亡,其机制主要与两者对细胞凋亡相关蛋白表达的调节有关。

关键词: 枸杞多糖, 顺铂, 肺癌, 抗氧化, 凋亡

Abstract: Objective: To investigate the effect of Lycium barbarum polysaccharides (LBP) combined with cisplatin (DDP) on oxidative injury, apoptosis and apoptosis-related protein expression in hunman lung adenocarcinoma cell line A549. Methods: The cells were divided into a control group, a DDP (6 mg/L) group, an LBP (8 mg/L) group and an LBP (8 mg/L) + DDP (6 mg/L) group. CCK-8 method was used to detect the effect of separate and combined treatment with LBP and DDP on the cell viability of A549. Superoxide dismutase activity was measured by xanthine oxidase method, malondialdehyde content by thiobarbituric acid method, the content of glutathione (GSH) by dithio-dinitrobenzoic acid method, the content of reactive oxygen species (ROS) and the rate of apoptosis by flow cytometry, and the expression of apoptosis-related proteins Bcl-2, Bax and caspase-3 by Western blot. Results: LBP significantly enhanced the inhibitory effect of DDP on the survival rate of A549 cells (P < 0.01). DDP resulted in a significant reduction of superoxide dismutase (SOD) activity and GSH content (P < 0.01), and a significant increase of MDA and ROS contents in A549 cells (P < 0.01). Compared with the DDP group, the levels of SOD, GSH and malondialdehyde in the LBP + DDP group were not significantly changed (P > 0.05), while the content of ROS was significantly reduced (P < 0.01).Treatment with DDP, LBP or their combination could significantly promote cell apoptosis (P < 0.01) and lower the expression of Bax and caspase-3 as well as Bax/Bcl-2 ratio (P < 0.01). Conclusion: The combination of LBP and DDP can obviously promote apoptosis of A549 cells, and its underlying mechanism is mainly related to the regulation of apoptosis-related protein expression by both of them.

Key words: Lycium barbarum polysaccharide, cisplatin, lung cancer, antioxidant, apoptosis

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