| 摘要: |
| 目的 探讨去氧地胆草素（Deo）对人非小细胞肺癌A549细胞中活性氧（ROS）调控细胞凋亡途径的影响。方法 用不同浓度的Deo分别作用于A549细胞，通过CCK-8法检测细胞活力，2’,7’-二氯荧光素二乙酸酯（DCFH-DA）染色检测细胞产生的ROS含量，Hoechst 33342染色检测细胞凋亡小体，流式细胞术检测细胞凋亡情况及线粒体膜电位，qPCR和蛋白质印迹法检测细胞凋亡相关蛋白表达。用A549细胞裸小鼠移植瘤模型评价Deo的体内抗肿瘤效果，通过H-E染色和TUNEL检测观察肿瘤组织坏死和细胞凋亡的变化。结果 随着Deo浓度（1、2、4、8、16、32 μmol/L）的增加，A549细胞活力呈现出递减趋势；10 μmol/L Deo可提高A549细胞内的ROS含量，降低线粒体膜电位，促进细胞凋亡；10、20 μmol/L Deo可促进A549细胞中Bax和caspase-3的mRNA及蛋白表达，而抑制Bcl-2的mRNA及蛋白表达；同时细胞质中细胞色素C表达明显增加。10、20 mg·kg－1·d－1 Deo给药17 d后，裸小鼠移植瘤的体积和重量均被抑制，并且肿瘤组织中坏死面积和细胞凋亡指数增大。结论 Deo能够诱导A549细胞产生ROS，激活线粒体凋亡通路，促进细胞凋亡。 |
| 关键词: 去氧地胆草素 肺肿瘤 A549细胞 活性氧 细胞凋亡 |
| DOI：10.16781/j.CN31-2187/R.20230236 |
| 投稿时间：2023-04-26修订日期：2023-10-26 |
| 基金项目:甘肃省卫生行业科研计划项目（GSWSKY-2014-45）. |
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| Deoxyelephantopin promotes apoptosis of human non-small cell lung cancer A549 cells by inducing reactive oxygen species |
| PAN Li1,ZHANG Yinfang1,QIU Yongyu1*,LIU Jinmei2 |
(1. Affiliated Hospital of Gansu University of Chinese Medicine, Lanzhou 730000, Gansu, China;
2. Family Planning Science and Technology Service Center of Gansu Province, Lanzhou 730000, Gansu, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect of deoxyelephantopin (Deo) on regulation of cell apoptosis pathway by reactive oxygen species (ROS) in human non-small cell lung cancer A549 cells. Methods A549 cells were treated with different concentrations of Deo. Cell counting kit 8 (CCK-8) assay was used to detect the cell viability, 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) staining was used to measure cellular ROS content, and Hoechst 33342 staining was used to detect apoptotic bodies. Cell apoptosis and mitochondrial membrane potential were examined by flow cytometry. Quantitative polymerase chain reaction and Western blotting were used to detect the expression of apoptosis-related proteins. A549 xenograft tumor model in nude mice was used to evaluate the anti-tumor effect of Deo in vivo. Hematoxylin-eosin staining and TUNEL assay were used to observe the necrosis and cell apoptosis in tumor tissues. Results With the increase of Deo concentration (1, 2, 4, 8, 16, 32 μmol/L), the viability of A549 cells showed a decreasing trend. Deo at 10 μmol/L could increase the ROS content in A549 cells, reduce mitochondrial membrane potential and promote cell apoptosis. Deo at 10 and 20 μmol/L promoted the mRNA and protein expression of Bax and caspase-3 and inhibited the mRNA and protein expression of Bcl-2, accompanied by significantly increased protein expression of cytochrome C in the cytoplasm. The volume and weight of transplanted tumor in nude mice were significantly inhibited after 17 d of administration of Deo of 10 and 20 mg·kg－1·d－1, and the necrotic area and the number of apoptotic cells in the tumor tissue were increased. Conclusion Deo can induce intracellular ROS production, and then activate the mitochondrial apoptosis pathway and consequently promote the apoptosis of A549 cells. |
| Key words: deoxyelephantopin lung neoplasms A549 cells reactive oxygen species apoptosis |
