一种优化设计的人工纳米材料遗传毒性评估方法及应用
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摘要
通过优化设计的蚕豆根尖微核技术监测了3种典型的人工纳米材料的遗传毒性,以期该方法可以作为人工纳米材料遗传毒性评估的实验教学方法,并能够推广使用。通过反复的实验,对传统的蚕豆根尖微核技术进行了优化设计,并用这种优化设计的蚕豆根尖微核技术对纳米铜、纳米氧化锌和纳米二氧化硅3种人工纳米材料的遗传毒性进行了评估。结果表明:(1)优化了传统的蚕豆根尖微核技术的关键步骤,如:震荡设置、染毒时根尖长度、解离、染色时间等;(2)3种纳米材料均能诱发较高的微核效应,且微核千分率比较:纳米铜>纳米氧化锌>纳米二氧化硅;(3)同种纳米材料不同浓度与阴性对照均有极显著性差异(p<0.01);同种浓度的3种不同纳米材料间的微核率有显著或者极显著差异(p<0.05或p<0.01),均有统计学意义;(4)采用优化的蚕豆根尖微核技术去评估人工纳米材料的遗传毒性具有很强的操作性和科学性。通过该文的研究,以期优化的蚕豆根尖微核技术能够得到更广的应用,并为今后纳米材料遗传毒性和安全性的评估提供依据和方法。
The genetic toxicity of three typical manufactured nanomaterials(MNMs) was monitored by using the optimally designed V. faba. root tip micronucleus technology. It is hoped that this method can be used as an experimental teaching method to assess genetic toxicity of MNMs, and can be employed widely. Through repeated experiments, the traditional V.faba. root tip micronucleus technology has been optimized. Thus it has been used to evaluate the genetic toxicity of three typical MNMs, such as nano-Cu, nano-ZnO and nano-SiO_2. The key steps of traditional V. faba. root tip micronucleus technology were optimized, such as shock setting, root tip length when exposed, dissociation, dyeing time, etc. Three kinds of MNMs all can induce higher micronucleus effect, and comparison result of micronucleus permillage is nano-Cu > nanoZnO > nano-SiO_2. The significance comparison between the same nanomaterial with different concentrations and negative control groups is extremely significant(p<0.01); the significance comparison between the three kinds of MNMs with same concentration is significant or extremely significant(p<0.05 or p<0.01). Both of them have statistical significance. It has stronger operability and scientificalness to evaluate genotoxicity of MNMs by using the optimized micronucleus test technique of V. faba. root tip cells. Through the study, it is expected that the optimized V. faba. root tip micronucleus technology can be applied more widely, and can provide the basis and method for the evaluation of genetic toxicity and safety of nanomaterials in the future.
The genetic toxicity of three typical manufactured nanomaterials(MNMs) was monitored by using the optimally designed V. faba. root tip micronucleus technology. It is hoped that this method can be used as an experimental teaching method to assess genetic toxicity of MNMs, and can be employed widely. Through repeated experiments, the traditional V.faba. root tip micronucleus technology has been optimized. Thus it has been used to evaluate the genetic toxicity of three typical MNMs, such as nano-Cu, nano-ZnO and nano-SiO_2. The key steps of traditional V. faba. root tip micronucleus technology were optimized, such as shock setting, root tip length when exposed, dissociation, dyeing time, etc. Three kinds of MNMs all can induce higher micronucleus effect, and comparison result of micronucleus permillage is nano-Cu > nanoZnO > nano-SiO_2. The significance comparison between the same nanomaterial with different concentrations and negative control groups is extremely significant(p<0.01); the significance comparison between the three kinds of MNMs with same concentration is significant or extremely significant(p<0.05 or p<0.01). Both of them have statistical significance. It has stronger operability and scientificalness to evaluate genotoxicity of MNMs by using the optimized micronucleus test technique of V. faba. root tip cells. Through the study, it is expected that the optimized V. faba. root tip micronucleus technology can be applied more widely, and can provide the basis and method for the evaluation of genetic toxicity and safety of nanomaterials in the future.
引文
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[13]谢海,袁翠平.应用蚕豆微核技术监测金沙江攀枝花市东区段水质[J].云南农业大学学报,2012,27(6):887-892.Xie Hai,Yuan Cuiping.Utilization of micronucleus test technique in Vicia faba root tips to monitor the water quality of Jinsha River in eastern section of Panzhihua[J].Journal of Yunnan Agricultural University,2012,27(6):887-892.
[14]国家环境保护局.环境监测技术规范-生物检测(水环境部分)[M].北京:中国环境科学出版社,1986.
[15]Na Bao,Yu Liu,Zi Weili,et al.Construction of order mesoporous(Eu-La)/ZnO composite material and its luminescent characters[J].Journal of Luminescence,2016(177):409-416.
[2]候俊,次翰林,吕博文,等.典型人工纳米材料的水环境行为研究进展[J].水资源保护,2017,36(6):1-6.Hou Jun,Ci Hanlin,Lyu Bowen,et al.Research progress of water environment behavior of typical engineered nanomaterials[J].Water Resources Protection,2017,36(6):1-6.
[3]Rmer I,White T A,Baalousha M,et a1.Aggregation and dispersion of silver nanoparticles in exposure media for aquatic toxicity tests[J].Journal of Chromatography A,2011,1218(27):4226-4233.
[4]Pfuhler S,Elespuru R,Aardema M J,et al.Genotoxicity of nanomaterials:refining strategies and tests for hazard identification[J].Environmental and Molecular Mutagenesis,2013,54(4):229-239.
[5]Doak S H,Dusinska M.NanoGenotoxicology:present and the future[J].Mutagenesis,2017,32(1):1-4.
[6]Jochums A,Friehs E,Sambale F,et al.Revelation of different nanoparticle-uptake behavior in two standard cell lines NIH/3T3 and A549 by flow cytometry and time-lapse imaging[J].Toxics,2017,5(3):110-115.
[7]罗勋.基于大肠杆菌-秀丽隐杆线虫二元食物链体系的纳米银生物有效性研究[D].合肥:中国科学技术大学,2017.Luo Xun.Bioavailability of Silver Nanoparticles Based on Food Chain from Esherichia coil to Caenorhabditis elegans[D].Hefei:University of Science and Technology of China,2017.
[8]赵佳琪,周燕,尹颖,等.纳米氧化锌暴露对沉水植物金鱼藻的毒性效应[J].南京大学学报:自然科学版,2017,53(5):894-902.Zhao Jiaqi,Zhou Yan,Yin Ying,et al.The toxic effects of zinc oxide nanoparticles exposure on submersed macrophyte Ceratophyllum demersum[J].Journal of Nanjing University:Natural Science Edition,2017,53(5):894-902.
[9]张洁,穆莉,胡献刚.纳米材料的食品暴露及其毒性研究进展[J].环境化学,2018,37(8):1770-1779.Zhang Jie,Mu Li,Hu Xiangang.Recent progress on the exposure and toxicity of nanoparticles in foods[J].Environmental Chemistry,2018,37(8):1770-1779.
[10]王秀娟,李婷竹,路强,等.纳米银和PVP包被纳米银对HepG2细胞遗传毒性的比较研究[J].生态毒理学报,2018,13(3):94-102.Wang Xiujuan,Li Tingzhu,Lu Qiang,et al.Comparative study on the genetic toxicity of nano silver and PVP coated silver nanoparticles to HepG2 cells[J].Asian Journal of Ecotoxicology,2018,13(3):94-102.
[11]文海若.适合纳米材料遗传毒性评价方法的选择[J].癌变·畸变·突变,2018,30(4):326-331.Wen Hairuo.The selection of adaptive evaluation method to nanomaterials toxicity[J].Carcinogenesis·Teratogenesis·Mutagenesis,2018,30(4):326-331.
[12]黄德超,任强,刘蓉,等.纳米银污染对蚕豆根尖微核率影响及评价研究[J].生态环境学报,2016,25(4):711-714.Huang Dechao,Ren Qiang,Liu Rong,et al.Research on the influence and evaluation of nano-silver pollution on the micronucleus rate of the root tip of Vicia faba[J].Ecology and Environmental Science,2016,25(4):711-714.
[13]谢海,袁翠平.应用蚕豆微核技术监测金沙江攀枝花市东区段水质[J].云南农业大学学报,2012,27(6):887-892.Xie Hai,Yuan Cuiping.Utilization of micronucleus test technique in Vicia faba root tips to monitor the water quality of Jinsha River in eastern section of Panzhihua[J].Journal of Yunnan Agricultural University,2012,27(6):887-892.
[14]国家环境保护局.环境监测技术规范-生物检测(水环境部分)[M].北京:中国环境科学出版社,1986.
[15]Na Bao,Yu Liu,Zi Weili,et al.Construction of order mesoporous(Eu-La)/ZnO composite material and its luminescent characters[J].Journal of Luminescence,2016(177):409-416.
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