李鹏, 赵敬利, 夏金童. 单壁碳纳米管诱导人支气管上皮细胞的氧化损伤和凋亡[J]. 环境与职业医学, 2013, 30(12): 942-946.
引用本文: 李鹏, 赵敬利, 夏金童. 单壁碳纳米管诱导人支气管上皮细胞的氧化损伤和凋亡[J]. 环境与职业医学, 2013, 30(12): 942-946.
LI Peng , ZHAO Jing-li , XIA Jin-tong . Single-Walled Carbon Nanotubes Induce Oxidative Stress and Apoptosis of Human Bronchial Epithelial Cell Line[J]. Journal of Environmental and Occupational Medicine, 2013, 30(12): 942-946.
Citation: LI Peng , ZHAO Jing-li , XIA Jin-tong . Single-Walled Carbon Nanotubes Induce Oxidative Stress and Apoptosis of Human Bronchial Epithelial Cell Line[J]. Journal of Environmental and Occupational Medicine, 2013, 30(12): 942-946.

单壁碳纳米管诱导人支气管上皮细胞的氧化损伤和凋亡

Single-Walled Carbon Nanotubes Induce Oxidative Stress and Apoptosis of Human Bronchial Epithelial Cell Line

  • 摘要: 目的 研究单壁碳纳米管(single-walled carbon nanotubes, SWCNTs)对人源支气管上皮细胞株(BEAS-2B)氧化应激和细胞凋亡的影响。

    方法 采用生长状态良好的BEAS-2B,分别以含0、10、50、100、200μg/mL SWCNTs的培养液处理24、48 h。采用四甲基偶氮唑盐(MTT)比色法检测SWCNTs对BEAS-2B细胞活力的影响;采用荧光探针2', 7'-二氯荧光素二乙酸(DCFH-DA)进行活性氧(ROS)检测;采用黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性;采用可见光法测定过氧化氢酶(CAT)活性;采用硫代巴比妥酸(TBA)法测定丙二醛(MDA)的含量;采用二硫代二硝基苯甲酸(DTNB)法测定谷胱甘肽过氧化物酶(GSH-Px)活性;采用阳离子荧光染料罗丹明123测定线粒体膜电位;采用pNA法检测细胞内caspase-3的蛋白活性; Real-time PCR测定细胞内baxbcl-2基因的表达变化。

    结果 当处理浓度大于10μg/mL时, SWCNTs可诱导BEAS-2B细胞存活率下降,细胞内ROS含量增加,线粒体膜电位减低, MDA升高,抗氧化酶系(SOD、CAT、GSH-Px)酶活性降低, caspase-3蛋白活性增加, bax在转录水平上表达量增加, bcl-2表达量降低,各实验结果均呈效应随浓度变化的趋势(P<0.05)。

    结论 SWCNTs能够诱导BEAS-2B细胞的氧化损伤和细胞凋亡。

     

    Abstract: Objective To investigate the effect of single-walled carbon nanotubes (SWCNTs) on oxidative stress and apoptosis of human bronchial epithelial cell line (BEAS-2B).

    Methods BEAS-2B cells were cultured with 0, 10, 50, 100, and 200 μg/mL SWCNTs separately for 24 h and 48 h. The viability of BEAS-2B cells was detected by tetrazolium (MTT) assay; the generation of reactive oxygen species (ROS) was detected by fluorescent probe (DCFH-DA), the enzymatic activity of super oxide dismutase (SOD) was detected by xanthine oxidase method; the enzymatic activity of catalase (CAT) was detected by visible spectrophotometry; the content of malondialdehyde (MDA) was determined by thiobarbituric acid (TBA); the enzymatic activity of glutathione peroxidase (GSH-Px) was determined by dithio nitrobenzoic acid (DTNB); the mitochondrial membrane potential was determined by cationic fluorescent dye rhodamine 123; the intracellular caspase-3 protein activity was determined by pNA assay; the mRNA levels of bax and bcl-2 were determined using reverse transtription polymerase chain reaction (RT-PCR).

    Results When the concentration of SWCNTs was more than 10 μg/mL, the viability of BEAS-2B cells declined, the levels of intracellular ROS increased, the mitochondrial membrane potential declined, the lipid peroxidation product (MDA) increased, the activities of antioxidant enzymes (SOD, CAT, GSH-Px) reduced, the protein activity of caspase-3 increased, the expression of bax mRNA in creased while the bcl-2 mRNA reduced. All the results were exhibited in a dose-response manner (P<0.05).

    Conclusion SWCNTs could induce oxidative stress and apoptosis in BEAS-2B cells.

     

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