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鸡血藤黄酮类有效部位对人肺腺癌A549细胞氧化应激的影响
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北京市教育委员会科技计划面上项目(KM200810025008)


Effect of Active Part of Spatholobus Suberectus Flavonoid on Oxidative Stress of Human Pulmonary Adenocarcinoma Cell A549
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    摘要:

    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。

    Abstract:

    Objective: To investigate the effect of active part of spatholobus suberectus flavonoid (SSCE) on oxidative stress in human pulmonary adenocarcinoma cell A549. Method: Cells were divided into a high-dose group, a moderate-dose,a low-dose group and a normal control group. Cell A549 were interfered with different dosages of SSCE active part for 24 h (normal group with DMEM) level of malonic dialde hyde(MDA) and glutathione(GSH); reactive oxygen speiesROS was detected using fluorescence microplate reader and confocal fluorescence microscope, and using DCFH-DA as fluorescent probe; Antioxidant was added, CCK-8 was employed to detect cell proliferation of A549 and ROS was detected by fluorescence microplate reader. Result: After the addition of SSCE, MDA content in each group increased (P<0.01) and GSH content in high density group decreased (P<0.01); Cell ROS content in high and middle density groups increased (P<0.01); Each group showed green fluorescence under confocal microscope, in which the high and middle density group showed higher; Addition of antioxidant made decrease of cell inhibition (P<0.05) and ROS content (P<0.01). Conclusion: SSCE can change the level of A549 MDA, GSH and ROS, and its mechanism of antitumor might be inducing oxidative stress reaction of tumor cells.

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林燕,李萍,王燕,解欣然,张蕾,何薇,杨国旺,王笑民.鸡血藤黄酮类有效部位对人肺腺癌A549细胞氧化应激的影响[J].中国实验方剂学杂志,2012,18(4):190~193

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