纳米硫化镉对A549细胞的损伤研究
|
摘要
研究纳米硫化镉(Nano-Cd S)材料对肺癌细胞系A549的毒性及氧化损伤作用。培养A549细胞,经传代后接种于6孔板中,每孔2 m L完全培养基,接种次日进行染毒。用直径20~30 nm、长度80~100 nm的Nano-Cd S进行染毒,染毒浓度分别为0、5、10、20、40和80 mg·L~(-1)。染毒24 h后用MTT检测细胞存活率,以存活率在80%左右的浓度为后续实验染毒浓度。应用流式细胞技术,用荧光探针法检测A549细胞的活性氧(reactive oxygen species,ROS)含量,PI-Annexin-V法检测细胞凋亡情况;用试剂盒检测细胞中超氧化物岐化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性以及丙二醛(malondialdehyde,MDA)含量,判断细胞氧化损伤情况。不同浓度Nano-Cd S处理细胞24 h之后,细胞存活率随剂量的增加而下降,浓度为10、20、40和80μg·L~(-1)时,存活率分别为(88.71%±0.80%)、(81.93%±3.06%)、(75.23%±1.13%)和(70.66%±5.63%),且各组间差异均具有统计学意义(P<0.05)。以浓度为10和20 mg·L~(-1)的Nano-Cd S染毒24 h后,胞内ROS含量和细胞凋亡率随染毒剂量的增加而增加(P<0.05);浓度为10 mg·L~(-1)时,细胞凋亡率为(6.26%±0.44%)。与对照相比,各染毒组SOD和CAT活性和MDA含量升高,20 mg·L~(-1)染毒组SOD和CAT活性和MDA含量高于10 mg·L~(-1)染毒组(P<0.05)。研究表明,纳米硫化镉能引起A549细胞的氧化损伤和细胞凋亡,具有明显的细胞毒性。
This paper aims to investigate the oxidative damage of the lung cancer cell line A549 following nanometer cadmium sulfide( Nano-Cd S) exposure. A549 cells were cultured in 6-well plates with 2 m L complete medium and then treated with Nano-Cd S for 24 h,whose diameters ranged from 20 to 30 nm and lengths were from 80 to100 nm. The doses of Nano-Cd S were 0,5,10,20,40 and 80 mg·L~(-1)respectively. MTT colorimetric assay was used to detect the cell survival rate,and the dose resulting in 80% cell survival rate was used as subsequent expo-sure dose. Flow cytometry was applied to detect the reactive oxygen species( ROS) contents of A549. The cell apoptosis was assayed by using PI-Annexin-V. The activities of SOD and CAT,the content of MDA were detected with the kits. After treatment with different concentrations of Nano-Cd S for 24 h,the survival rates of cells were( 88. 71% ± 0. 80%),( 81. 93% ± 3. 06%),( 75. 23% ± 1. 13%) and( 70. 66% ± 5. 63%) respectively at doses of 10,20,40 and 80 mg·L~(-1),showing a signficant decrease( P < 0. 05) with Nano-Cd S dose increase. Intracellular ROS contents and apoptosis rate of cells increased significantly after being treated with 10 or 20 mg·L~(-1)Nano-Cd S,compared with the control( P < 0. 05). The apoptosis rates were( 6. 26% ± 0. 44%) and( 8. 94% ±1. 38%) at 10 and 20 mg·L~(-1)Nano-Cd S groups. Compared with control,the activities of SOD and CAT,and the content of MDA all increased. The activities of SOD and CAT,and the content of MDA in 20 mg·L~(-1)exposure group were significantly higher compared to 10 mg·L~(-1)exposure group( P < 0. 05). It is indicated that Nano-Cd S exposure can cause oxidative damage and apoptosis in A549 cells and then result in cytotoxicity.
This paper aims to investigate the oxidative damage of the lung cancer cell line A549 following nanometer cadmium sulfide( Nano-Cd S) exposure. A549 cells were cultured in 6-well plates with 2 m L complete medium and then treated with Nano-Cd S for 24 h,whose diameters ranged from 20 to 30 nm and lengths were from 80 to100 nm. The doses of Nano-Cd S were 0,5,10,20,40 and 80 mg·L~(-1)respectively. MTT colorimetric assay was used to detect the cell survival rate,and the dose resulting in 80% cell survival rate was used as subsequent expo-sure dose. Flow cytometry was applied to detect the reactive oxygen species( ROS) contents of A549. The cell apoptosis was assayed by using PI-Annexin-V. The activities of SOD and CAT,the content of MDA were detected with the kits. After treatment with different concentrations of Nano-Cd S for 24 h,the survival rates of cells were( 88. 71% ± 0. 80%),( 81. 93% ± 3. 06%),( 75. 23% ± 1. 13%) and( 70. 66% ± 5. 63%) respectively at doses of 10,20,40 and 80 mg·L~(-1),showing a signficant decrease( P < 0. 05) with Nano-Cd S dose increase. Intracellular ROS contents and apoptosis rate of cells increased significantly after being treated with 10 or 20 mg·L~(-1)Nano-Cd S,compared with the control( P < 0. 05). The apoptosis rates were( 6. 26% ± 0. 44%) and( 8. 94% ±1. 38%) at 10 and 20 mg·L~(-1)Nano-Cd S groups. Compared with control,the activities of SOD and CAT,and the content of MDA all increased. The activities of SOD and CAT,and the content of MDA in 20 mg·L~(-1)exposure group were significantly higher compared to 10 mg·L~(-1)exposure group( P < 0. 05). It is indicated that Nano-Cd S exposure can cause oxidative damage and apoptosis in A549 cells and then result in cytotoxicity.
引文
[1]常元勋.金属毒理学[M].北京:北京大学医学出版社,2008
[2]Choi A O,Cho S J,Desbarats J,et al.Quantum dotinduced cell death involves Fas upregulation and lipid peroxidation in human neuroblastoma cells[J].Journal of Nanobiotechnology,2007,5(1):1-13
[3]李云蕊,李大鸣,何宁,等.纳米硫化镉对大鼠肾小管上皮细胞抗氧化酶的影响[J].环境与健康杂志,2011,28(10):867-870Li Y R,Li D M,He N,et al.Effects of cadmium sulfide nano-particles on antioxidases in TECs cells[J].Journal of Environment and Health,2011,28(10):867-870(in Chinese)
[4]周庆红,姜淑卿,刘英华,等.纳米硫化镉与常规硫化镉对雄性小鼠的生殖毒性[J].环境与健康杂志,2014,31(4):299-301Zhou Q H,Jiang S Q,Liu Y H,et al.Reproductive toxicity of nano-cadmium sulfide and normal-sized cadmium sulfide on male mice[J].Journal of Environment and Health,2014,31(4):299-301(in Chinese)
[5]梁丽君,张静姝,郭昌胜,等.硫化镉量子点对人胚肝细胞L-02的毒性研究[J].生态毒理学报,2010,5(4):525-530Liang L J,Zhang J S,Guo C S,et al.Cadmium sulfide quantum dots induce cytotoxicity in human embryo liver cells[J].Asian Journal of Ecotoxicology,2010,5(4):525-530(in Chinese)
[6]张静姝,柏杉山,刘英华,等.硫化镉量子点对中国仓鼠肺成纤维细胞损伤的实验研究[J].中华劳动卫生职业病杂志,2010,28(1):39-41
[7]张浩,黄新杰,刘秀玉,等.纳米材料安全性的研究进展及其评价体系[J].过程工程学报,2013,13(5):893-899
[8]Cornejo-Garrido H,Kibanova D,Nieto-Camacho A,et al.Oxidative stress,cytoxicity,and cell mortality induced by nano-sized lead in aqueous suspensions[J].Chemosphere,2011,84(10):1329–1335
[9]郑国颖,李茂静,郝玉兰,等.不同粒径和形貌的纳米硫化镉对小鼠肝肾毒性的研究[J].环境与健康杂志,2010,27(6):542-543
[10]刘佳蕙,苏敏,孙可欣,等.镉系量子点的细胞毒性——镉离子之辩[J].西南民族大学学报,2015,41(3):310-315Liu J H,Su M,Sun K X,et al.Cytotoxicity of cadmiumcontaining quantum dots—Debate on releasing cadmium ions to induce cytotoxicity[J].Journal of Southwest University for Nationalities:Natural Science Edition,2015,41(3):310-315(in Chinese)
[11]余强,李坤刚,文兴,等.纳米硫化镉量子点细胞毒性作用机制[J].生态毒理学报,2009,4(4):488-493Yu Q,Li K G,Wen X,et al.The cytotoxicity mechanism of unmodified cadmium sulfide quantum dots[J].Asian Journal of Ecotoxicology,2009,4(4):488-493(in Chinese)
[12]李亦白,鲍一苓,樊佳,等.碲化镉量子点对A549细胞的毒性研究[J].现在中西医结合杂志,2014,23(18):1950-1943Li Y B,Bao Y L,Fan J,et al.Study on cytotoxicity of cadmium telluride quantum dot on A549 cells[J].Modern Journal of Integrated Traditional Chinese and Western Medicine,2014,23(18):1950-1943(in Chinese)
[2]Choi A O,Cho S J,Desbarats J,et al.Quantum dotinduced cell death involves Fas upregulation and lipid peroxidation in human neuroblastoma cells[J].Journal of Nanobiotechnology,2007,5(1):1-13
[3]李云蕊,李大鸣,何宁,等.纳米硫化镉对大鼠肾小管上皮细胞抗氧化酶的影响[J].环境与健康杂志,2011,28(10):867-870Li Y R,Li D M,He N,et al.Effects of cadmium sulfide nano-particles on antioxidases in TECs cells[J].Journal of Environment and Health,2011,28(10):867-870(in Chinese)
[4]周庆红,姜淑卿,刘英华,等.纳米硫化镉与常规硫化镉对雄性小鼠的生殖毒性[J].环境与健康杂志,2014,31(4):299-301Zhou Q H,Jiang S Q,Liu Y H,et al.Reproductive toxicity of nano-cadmium sulfide and normal-sized cadmium sulfide on male mice[J].Journal of Environment and Health,2014,31(4):299-301(in Chinese)
[5]梁丽君,张静姝,郭昌胜,等.硫化镉量子点对人胚肝细胞L-02的毒性研究[J].生态毒理学报,2010,5(4):525-530Liang L J,Zhang J S,Guo C S,et al.Cadmium sulfide quantum dots induce cytotoxicity in human embryo liver cells[J].Asian Journal of Ecotoxicology,2010,5(4):525-530(in Chinese)
[6]张静姝,柏杉山,刘英华,等.硫化镉量子点对中国仓鼠肺成纤维细胞损伤的实验研究[J].中华劳动卫生职业病杂志,2010,28(1):39-41
[7]张浩,黄新杰,刘秀玉,等.纳米材料安全性的研究进展及其评价体系[J].过程工程学报,2013,13(5):893-899
[8]Cornejo-Garrido H,Kibanova D,Nieto-Camacho A,et al.Oxidative stress,cytoxicity,and cell mortality induced by nano-sized lead in aqueous suspensions[J].Chemosphere,2011,84(10):1329–1335
[9]郑国颖,李茂静,郝玉兰,等.不同粒径和形貌的纳米硫化镉对小鼠肝肾毒性的研究[J].环境与健康杂志,2010,27(6):542-543
[10]刘佳蕙,苏敏,孙可欣,等.镉系量子点的细胞毒性——镉离子之辩[J].西南民族大学学报,2015,41(3):310-315Liu J H,Su M,Sun K X,et al.Cytotoxicity of cadmiumcontaining quantum dots—Debate on releasing cadmium ions to induce cytotoxicity[J].Journal of Southwest University for Nationalities:Natural Science Edition,2015,41(3):310-315(in Chinese)
[11]余强,李坤刚,文兴,等.纳米硫化镉量子点细胞毒性作用机制[J].生态毒理学报,2009,4(4):488-493Yu Q,Li K G,Wen X,et al.The cytotoxicity mechanism of unmodified cadmium sulfide quantum dots[J].Asian Journal of Ecotoxicology,2009,4(4):488-493(in Chinese)
[12]李亦白,鲍一苓,樊佳,等.碲化镉量子点对A549细胞的毒性研究[J].现在中西医结合杂志,2014,23(18):1950-1943Li Y B,Bao Y L,Fan J,et al.Study on cytotoxicity of cadmium telluride quantum dot on A549 cells[J].Modern Journal of Integrated Traditional Chinese and Western Medicine,2014,23(18):1950-1943(in Chinese)