食品科学 ›› 2019, Vol. 40 ›› Issue (13): 110-115.doi: 10.7506/spkx1002-6630-20180707-093

• 营养卫生 • 上一篇    下一篇

苦荞茶多糖诱导人肺腺癌A549细胞凋亡的线粒体机制

杨二万,张 敏,杨兴斌,杨红燕   

  1. 1.空军军医大学航空航天医学系,陕西 西安 710032;2.陕西师范大学食品工程与营养科学学院,陕西 西安 710119
  • 出版日期:2019-07-15 发布日期:2019-07-23
  • 基金资助:
    国家自然科学基金面上项目(81470412)

Tartary Buckwheat Tea Polysaccharide Induces Mitochondria-Mediated Apoptosis in A549 Human Lung Cancer Cells

YANG Erwan, ZHANG Min, YANG Xingbin, YANG Hongyan   

  1. 1. Department of Aerospace Medicine, Air Force Medical University, Xi’an 710032, China; 2. College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an 710119, China
  • Online:2019-07-15 Published:2019-07-23

摘要: 目的:从线粒体调控机制的角度探讨苦荞茶多糖(tartary buckwheat tea polysaccharide,TBTP)对人肺腺癌A549细胞凋亡的影响。方法:应用水提醇沉法结合超声辅助技术提取的TBTP处理A549细胞,分为空白对照组和不同质量浓度TBTP处理组。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法和流式细胞术检测细胞增殖和凋亡水平,采用DHE和Mito SOX染色检测细胞和线粒体活性氧簇水平,采用JC-1染色检测线粒体膜电位变化,采用Western blot法检测凋亡相关蛋白Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、Bcl-2、剪切型半胱氨酸蛋白酶(Caspase)-3、细胞色素c的水平。结果:TBTP呈剂量和时间依赖性抑制A549细胞增殖。TBTP处理A549细胞24 h可呈剂量依赖性地促进细胞凋亡。TBTP处理能够显著增加A549细胞和线粒体内活性氧水平,降低线粒体膜电位;同时,TBTP能够增加促凋亡蛋白Bax和剪切型Caspase-3的表达、抑制抗凋亡蛋白Bcl-2的表达,促进A549细胞线粒体细胞色素c外流。结论:TBTP能够促进活性氧簇生成、降低线粒体膜电位,进而抑制A549细胞增殖,促进细胞凋亡。

关键词: 苦荞茶, 多糖, 肺癌, 细胞凋亡

Abstract: Objective: To investigate the effect of tartary buckwheat tea polysaccharide (TBTP) on the apoptosis of A549 human lung cancer cells and its underlying mechanisms. Methods: TBTP was obtained by water extraction. 3-(4,5-Dimethyl- 2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to detect the influence of TBTP on the proliferation of A549 cells. Flow cytometry was applied to determine the effect of TBTP on cell apoptosis in A549 cells. The production of reactive oxygen species (ROS) was assessed by DHE and Mito SOX Red staining. The change of mitochondrial membrane potential was assessed by JC-1 staining. Meanwhile, Western blot analysis was used to evaluate the expression of Bcl-2-associated X protein (Bax), Bcl-2, cleaved caspase-3 and cytochrome c. Results: MTT assay showed a significant inhibitory effect of TBTP on A549 cells proliferation in a dose- and time-dependent manner. After 24 hours’ cultivation, TBTP also promoted A549 cells apoptosis in a dose-dependent manner. TBTP treatment promoted mitochondrial ROS production and oxidative stress in A549 cells. The mitochondrial membrane potential of A549 cells also decreased after TBTP administration, accompanied with increased cytochrome c release from mitochondria. Furthermore, TBTP treatment up-regulated the expression of Bax and cleaved caspase-3, and decreased Bcl-2 expression. Conclusion: TBTP exerts a novel anti-proliferative effect through induction of mitochondria-mediated apoptosis in A549 cells.

Key words: tartary buckwheat tea, polysaccharide, lung cancer, apoptosis

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