毛兰素通过ROS/p38 MAPK通路诱导人肺癌A549细胞凋亡  

Erianin induces apoptosis of human lung cancer A549 cells via ROS/p38 MAPK pathway

作　　者：邓同兴[1] 王敏丽[2] 温文静 袁磊[1] DENG Tong-xing;WANG Min-li;WEN Wen-jing;YUAN Lei(Department of Basic Medicine, Luohe Medical College, Luohe 462002 , China;Department of Pediatrics, The Second Affiliated Hospital of Luohe Medical College, Luohe 462300 , China)

机构地区：[1]漯河医学高等专科学校基础医学部,河南漯河462002 [2]漯河医学高等专科学校第二附属医院儿科,河南漯河462300

出　　处：《中国病理生理杂志》2019年第8期1457-1462,共6页Chinese Journal of Pathophysiology

基　　金：河南省科技厅科技发展计划项目(No.142102310203);漯河医学高等专科学校研究计划项目(No.2014-S-LMC01)

摘　　要：目的:研究毛兰素(erianin)对人肺癌A549细胞活力和凋亡的影响,并探讨可能的分子机制。方法:常规培养人肺癌A549细胞和人正常支气管上皮BEAS-2B细胞,给予不同浓度(0、10、20、40、80和160 nmol/L)毛兰素处理48 h后,采用CCK-8法检测细胞活力,流式细胞术检测活性氧(ROS)含量和细胞凋亡情况,WST-8法检测超氧化物歧化酶(SOD)活性,硫代巴比妥酸法检测丙二醛(MDA)含量,Western blot法检测核因子E2相关因子2(Nrf2)、NAD(P)H:醌氧化还原酶1(NQO1)和血红素氧合酶1(HO-1)蛋白的表达以及p38 MAPK的磷酸化和caspase-3蛋白的活化。结果:毛兰素对A549细胞活力具有较强的抑制作用(P<0.05),并呈剂量依赖性,IC50为52.64 nmol/L;毛兰素还可剂量依赖性地诱导细胞凋亡(P<0.05),显著升高ROS和MDA含量(P<0.05),抑制SOD活性(P<0.05),下调Nrf2、NQO1和HO-1蛋白表达水平(P<0.05);此外,毛兰素可上调p38 MAPK的磷酸化水平并激活caspase-3(P<0.05),该作用可被抗氧化剂N-乙酰-L-半胱氨酸和p38 MAPK抑制剂SB203580显著抑制(P<0.05)。结论:毛兰素可在体外诱导人肺癌A549细胞凋亡,这可能与其抑制SOD活性,下调Nrf2、NQO1和HO-1蛋白表达,继而导致ROS含量升高和激活p38 MAPK有关。AIM: To investigate the effect of erianin on the viability and apoptosis of human lung cancer A549 cells and its possible mechanism. METHODS: A549 cells and BEAS-2 B cells were cultured in vitro and treated with erianin at 10, 20, 40, 80 and 160 nmol/L for 48 h. The cell viability was measured by CCK-8 assay. The apoptosis and reactive oxygen species(ROS) were analyzed by flow cytometry. The activity of superoxide dismutase(SOD) was detected by WST-8 method, and the content of malondialdehyde(MDA) was detected by barbituric acid method. The protein levels of nuclear factor E2-related factor 2(Nrf2), NAD(P)H:quinone oxidoreductase-1(NQO1), heme oxygenase-1(HO-1), p38 MAPK, p-p38 MAPK, caspase-3 and cleaved caspase-3 were determined by Western blot.RESULTS: Erianin remarkably reduced the viability of A549 cells in a dose-dependent manner(P<0.05) with IC50 at 52.64 nmol/L. Erianin also induced apoptosis(P<0.05), increased ROS level and MDA content(P<0.05), diminished SOD activity(P<0.05), and down-regulated the protein levels of Nrf2, NQO1 and HO-1(P<0.05), in a dose-dependent manner. Meanwhile, erianin up-regulated the levels of p-p38 MAPK and cleaved caspase-3(P<0.05), and these effects were inhibited by N-acetyl-L-cysteine and SB203580(P<0.05).CONCLUSION: Erianin may induce apoptosis of human lung cancer A549 cells most likely via inhibiting SOD activity and down-regulating the protein levels of Nrf2, NQO1 and HO-1, thus resulting in an increase in ROS and activation of p38 MAPK.

关 键 词：毛兰素 肺癌 活性氧 细胞凋亡 P38 MAPK信号通路 

分 类 号：R734.2[医药卫生—肿瘤] R730.23[医药卫生—临床医学]