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目的:探讨鸡血藤黄酮类有效部位(SSCE)对人肺腺癌A549细胞的杀伤活性与氧化应激反应的相关性。方法:实验分为SSCE高、中、低剂量组和正常对照组,各组分别加入不同浓度的SSCE(正常组用培养基代替)作用人肺腺癌A549细胞24 h后,采用脂质过氧化物(MDA)、谷胱甘肽(GSH)试剂盒检测A549细胞MDA,GSH的含量;以2’,7’-二氯荧光素二乙酸酯(DCFH-DA)为荧光探针,用荧光酶标法和共聚焦荧光显微镜法检测活性氧(ROS);选择加入SSCE高浓度组同时加入抗氧化剂维生素C(Vit C),维生素E(Vit E),谷胱甘肽过氧化物酶(GPx),过氧化氢酶(CAT)作用细胞24 h后,采用CCK-8法检测A549细胞增殖情况及荧光酶标仪法检测ROS。结果:加入SSCE后,SSCE各组细胞MDA均含量上升(P<0.01)、高浓度组GSH含量下降(P<0.01);高、中浓度组细胞ROS含量上升(P<0.01);共聚焦显示SSCE各组均有绿色荧光,其中高、中浓度组荧光较强;加入抗氧化剂后,各组细胞抑制率均降低(P<0.05),ROS含量均下降(P<0.01)。结论:SSCE可以改变A549细胞MDA,GSH,ROS水平,其抗肿瘤作用可能是通过诱导肿瘤细胞氧化应激反应而实现的。
Objective: To investigate the correlation between the cytotoxic activity of SSCE on human lung adenocarcinoma A549 cells and oxidative stress. Methods: The experiment was divided into SSCE high, medium and low dose groups and normal control group, each group were added with different concentrations of SSCE (normal group replaced by medium) human lung adenocarcinoma A549 cells for 24 h, using lipid peroxidation (MDA) and glutathione (GSH) were used to detect the contents of MDA and GSH in A549 cells. Fluorescence was detected by fluorescence with 2 ’, 7’- dichlorofluorescein diacetate (DCFH-DA) Reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay (ELISA) and confocal fluorescence microscopy. Antioxidant vitamins C, Vit E, GPx ), Catalase (CAT) -treated cells for 24 h, the proliferation of A549 cells was detected by CCK-8 assay and ROS was detected by fluorescence microplate reader. Results: The content of MDA in the cells of SSCE group increased (P <0.01) and the content of GSH in high concentration group decreased (P <0.01), and the ROS content of cells in high and medium concentration groups increased (P <0.01) Each group of SSCE had green fluorescence, of which the fluorescence intensity was high in high and medium concentration groups. After addition of antioxidants, the inhibition rate of cells in each group decreased (P <0.05) and ROS content decreased (P <0.01). Conclusion: SSCE can change the level of MDA, GSH and ROS in A549 cells. The anti-tumor effect of SSCE may be through the induction of oxidative stress in tumor cells.