食品科学 ›› 2017, Vol. 38 ›› Issue (19): 177-181.doi: 10.7506/spkx1002-6630-201719028

• 营养卫生 • 上一篇    下一篇

低聚葡萄籽原花青素联合顺铂对人肺腺癌细胞A549凋亡的影响

王 海,连燕娜,周 游,黄姗婉,高丽萍*   

  1. 北京联合大学应用文理学院,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:2017-10-15 发布日期:2017-09-29
  • 基金资助:
    北京市自然科学基金项目(7163211)

Effect of Oligomeric Grape Seed Proanthocyanidins Combined with Cisplatin on the Apoptosis of Human Lung Adenocarcinoma Cell Line A549

WANG Hai, LIAN Yanna, ZHOU You, HUANG Shanwan, GAO Liping*   

  1. Beijing Key Laboratory of Bioactive Substances and Functional Foods, College of Arts and Sciences, Beijing Union University, Beijing 100191, China
  • Online:2017-10-15 Published:2017-09-29

摘要: 目的:研究低聚葡萄籽原花青素(oligomeric grape seed proanthocyanidins,O-GSP)联合顺铂对人肺腺 癌细胞A549抗氧化、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、顺铂(5 mg/L)组、O-GSP (4 mg/L)组、O-GSP(4 mg/L)+顺铂(5 mg/L)组。硫代巴比妥酸法测定细胞中丙二醛(malondialdehyde, MDA)含量;二硫代二硝基苯甲酸法测定谷胱甘肽(glutathione,GSH)含量;黄嘌呤氧化酶法测定超氧化物歧 化酶(superoxide dismutase,SOD)活力;流式细胞术检测细胞内活性氧(reactive 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:顺铂导致A549细胞MDA、 ROS含量显著升高(P<0.05),GSH含量和SOD活力显著降低(P<0.05),O-GSP对细胞内MDA含量、GSH含 量、SOD活力无显著影响(P>0.05)。O-GSP+顺铂组相对于顺铂组,细胞内MDA、GSH、SOD水平无显著性变化 (P>0.05),而ROS含量明显降低(P<0.05)。顺铂、O-GSP以及两者联用均能显著促进细胞凋亡(P<0.05),并且能 够显著提高Bax及Bax/Bcl-2值,顺铂和O-GSP+顺铂可显著降低Bcl-2的表达(P<0.05),同时激活caspase-3。结论: O-GSP联合顺铂可促进A549细胞的凋亡,其机制与O-GSP的抗氧化作用没有显著联系,而是与两者可共同调节凋亡相 关蛋白基因表达有关。

关键词: 低聚葡萄籽原花青素, 顺铂, 肺腺癌, 细胞凋亡

Abstract: Objective: To study the effect of oligomericgrape seed proanthocyanidin (O-GSP) combined with cisplatin (CDDP) on antioxidant defense systems, cell apoptosis and apoptosis-related protein expression in human lung adenocarcinoma cell line A549. Methods: The cells were divided into control, CDDP (5 mg/L), O-GSP (4 mg/L), and O-GSP (4 mg/L) + CDDP (5 mg/L) groups. Malondialdehyde (MDA) content was measured by thiobarbituric acid method. Glutathione (GSH) content was determined by nitrobenzoic acid method. Superoxide dismutase (SOD) activity was examined by xanthine oxidase method. Flow cytometry was used to detect the intracellular ROS content and apoptosis rate. The expression of apoptosis-related proteins such as Bcl-2, Bax and caspase-3 were detected by Western blot. Results: CDDP resulted in a significant increase of MDA and ROS in A549 cells (P < 0.05) and a significant reduction of GSH content and SOD activity (P < 0.05), but O-GSP had no significant effect on the levels of intracellular MDA, GSH and SOD (P > 0.05). Compared with the CDDP group, the levels of MDA, GSH and SOD in the O-GSP + CDDP group were not significantly changed (P > 0.05), while the content of ROS was significantly reduced (P < 0.05). CDDP, O-GSP and their combination could significantly promote cell apoptosis (P < 0.05). The combination of CDDP and O-GSP could increase Bax level and the ratio of Bax to Bcl-2, decrease Bcl-2 expression (P < 0.05), and activate caspase-3. Conclusion: Oligomeric grape seed proanthocyanidins combined with cisplatin can promote the apoptosis of A549 cells, and the apoptosis mechanism was not significantly related to the antioxidant activity of oligomeric grape seed proanthocyanidins but related to the expression of apoptosis-related genes.

Key words: oligomeric grape seed proanthocyanidin (O-GSP), cisplatin, lung adenocarcinoma, cell apoptosis

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