Microsoft Word 缪文彬-二校 190

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1 7 1 Vol. 7 No Journal of Food Safety and Quality Jan., 2016 缪文彬 *, 蒋伟, 陈相 (, ) 摘要 : 目的, 方法 28, 结果 mg/ml mg/ml hprt, mg/ml hprt (P<0.05), hprt (P<0.01) 结论 28 关键词 : ; ; ; Effect of fluorescent brightener on micronucleus and gene mutation of Chinese hamster ovary cells MIAO Wen-Bin *, JIANG Wei, CHEN Xiang (Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai , China) ABSTRACT: Objective To study the effect of fluorescent brightener on micronucleus and gene mutation of Chinese hamster ovary cells (CHO), and to discuss the mechanism of genetic damage of mammalian cell in vitro. Methods The genetic damage of CHO cells treated with different concentrations of fluorescent brightener 28 was detected with the method of mammalian cell micronucleus test and mammalian cell gene mutation test in vitro. Results There was no significant difference of hprt gene mutation frequencies and micronucleus rates between negative control group and mg/ml and mg/ml fluorescent brightener 28 treated groups. The hprt gene mutation frequency and micronucleus rate of CHO cells with mg/ml fluorescent brightener 28 were significant higher than that of negative control group (P<0.05). The hprt gene mutation frequency and micronucleus rate of CHO cells in positive control group were significant higher than that of negative control group (P<0.01). Conclusion Fluorescent brightener 28 may induce genetic damage in cultured mammalian cells. KEY WORDS: fluorescent brightener; micronucleus; gene mutation; genetic damage 基金项目 : (2011IK042) Fund: Supported by Science and Technology Project of General Administration of Quality Supervision, Inspection and Quarantine (2011IK042) * 通讯作者 :,, miuwb@shciq.gov.cn *Corresponding author: MIAO Wen-Bin, Engineer, Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai , China. miuwb@shciq.gov.cn

2 1, : 引言, 220 (V79), [1] [2,3] Ames [4],, [5],,,,,,,, [6],,, 2 材料与方法 2.1 材料 28(Sigma ), >99%, 100 mg (dimethyl sulfoxide, DMSO) 2 ml 50 mg/ml, DMSO DMSO, C(sigma ), 50 mg C, DMSO 2 ml 7,12-, 25 mg 7,12-, DMSO 5 ml 2.2 靶细胞 OECD CHO-k1,,, CHO,, 100 mm,,, 10 ml 37 CO 2 24 h [7] 2.3 四甲基偶氮噻唑蓝 (MTT) 实验 CHO 28, MTT 4 h, DMSO, OD 570 IC 50, g/ml, 60%, 5 mg/ml,, mg/ml,, 4 h 50 L MTT,, 150 L DMSO, 5 min,,, 2.4 体外哺乳动物细胞微核实验 24 h, 0.1 ml (100 ) 10 ml, 6 h,, 5 ml 2, 10 ml, CO 2, 18 h,,, (1500 r/min, 5 min),, mol/l 30 min, / 2, 15 min 1000,, ( ) [8] 2.5 体外哺乳动物细胞基因突变实验 24 h,, 0.1 ml (100 ) 2 ml S9, 10 ml, 4 h,, 5 ml 2, 10 ml, CO 2, 20 h

3 190 7,,, 100 mm 200, 3, d, Giemsa, 9 d, 6- (6-thioguanine, 6-TG),,,, 6-TG, 3, d, Giemsa, 6-TG( 10 g/ml), 3, 12 d Giemsa,, hprt [9] 2.6 统计分析 Excel, SPSS 12.0( SPSS ) hprt α= 结果与分析 3.1 荧光增白剂细胞毒性情况 28 ( 70%) 1 表 1 荧光增白剂细胞毒性 ( x ±SD) Table 1 Cytotoxicity of fluorescent brightener ( x ±SD) (mg/ml) (%) mg/ml ± g/ml ± g/ml ± ± 荧光增白剂致体外哺乳动物细胞微核数比较 (P<0.01),, m/ml mg/ml, mg/ml (P<0.05), mg/ml 28,,, - 表 2 荧光增白剂致体外哺乳动物细胞微核数 ( x ±SD) Table 2 Effect of micronucleus induced by fluorescent brightener in CHO cells ( x ±SD) (mg/ml) ( ) ± mg/ml ± mg/ml ± mg/ml ±0.77 * 31.44±0.62 ** :, * P<0.05, ** P< 荧光增白剂致中国仓鼠卵巢细胞基因集落形成效率和突变频率, hprt (P<0.01), 3, mg/ml mg/ml hprt, mg/ml hprt (P<0.05) 28 CHO hprt 表 3 荧光增白剂致中国仓鼠卵巢细胞基因突变作用 ( x ±SD) Table 3 Effect of gene mutation induced by fluorescent brightener in CHO cells ( x ±SD) (%) ( 10-5 ) 64.2± ± mg/ml 72.8± ± mg/ml 46.2± ± mg/ml 75.0± ± (5 g/ml) 60.0± ±4.5 <0.001 : * P 4 讨论,, DNA P *

4 1, : 191 [10] hprt hprt,, hprt 6-TG,,, [11,12],,,, DNA, [13,14], mg/ml 28, mg/ml mg/ml 28, 28 CHO, mg/ml mg/ml hprt, mg/ml hprt hprt, hprt,, [15] 28, 28 hprt,, 28 CHO, 28 CHO, 参考文献 [1],,. 220 [J]., 2015, 27(2): 1 9. Dong ZS, Wang ZY, Dong SH. Toxicological data and safety of fluorescent whitening agent 220 [J]. Paper Chem, 2015, 27(2): 1 9. [2],,. [J]., 2007, 4: Guo HP, Zhang MY, Liu YH. Analysis on the toxin of fluorescent whitening agents [J]. Hunan Papermaking, 2007, 4: [3]. CXT [J]., 2012, 4: Dong ZS. The safety of the use of fluorescent whitening agent CXT in detergent according the toxicological data [J]. China Cleaning Ind, 2012, 4: [4],,,. [J]., 1999, 11(11): Zhong WJ, Dong MZ, Xiao P, et al. Study on phototoxicity, photoallergy and mutagenicity of fluorescent whitening agents (FWAs) used in toilet paper [J]. Shanghai J Pre Med, 1999, 11(11): [5] Jung H, Seok SH, Han JH, et al. Effect of fluorescent whitening agent on the transcription of cell damage-related genes in zebrafish embryos [J]. J Appl Toxicol, 2012, 32(9): [6],. [J]., 2002, 25(8): 5 8. Shen YJ, Xu X. Toxic properties of fluorescent whitening agents [J]. Chem Technol Market, 2002, 25(8): 5 8. [7],. MTT Cr(Ⅵ) [J]., 2000, 27(3): 49. Jia G, Liu SJ. Application of trypan blue exclusion method and MTT method to determine Cr(Ⅵ)cell toxicity [J]. China Occopat Med, 2000, 27(3): 49. [8] Organization for Economic Cooperation and Development. In vitro mammalian cell micronucleus test [R]. Pairs: OECD, [9] Organization for Economic Cooperation and Development. In vitro mammalian cell gene mutation test [R]. Pairs: OECD, [10] Iarmarcovai G, Ceppi M, Botta A, et al. Micronuclei frequency in peripheral blood lymphocytes of cancer patients: a meta-analysis [J]. Mutat Res-Rev Mutat, 2008, 659(3): [11],,,. hprt [J]., 2002, 28(3): Zhou JH, Xue L, Du J, et al. Study on the frequency of Cd and HgCl 2 induced hprt locus mutation and its dose-effect relationship [J]. Ind Health Occupat Dis, 2002, 28(3): [12],,,. hprt [J]., 2001, 27(1):

192 7 23 25. Xue L, Zhou JH, Wang K, et al. Study on the frequency of HgCl 2 induced hprt locus mutation and its dose-effect relationship [J]. Ind Health Occupat Dis, 2001, 27(1): 23 25.

5 Xue L, Zhou JH, Wang K, et al. Study on the frequency of HgCl 2 induced hprt locus mutation and its dose-effect relationship [J]. Ind Health Occupat Dis, 2001, 27(1): [13] Terradas M, Martín M, Tusell L, et al. DNA lesions sequestered in micronuclei induce a local defective-damage response [J]. DNA Repair, 2009, 8(10): [14] Chang P, Li Y, Li D. Micronuclei levels in peripheral blood lymphocytes as a potential biomarker for pancreatic cancer risk [J]. Carcinogenesis, 2011, 32(2): [15] Muir P, Osborne Y, Morley AA, et al. Karyotypic abnormality of the X chromosome is rare in mutant HPRT-lymphocyte clones [J]. Mutation Research-fundamental and Molecular Mechanisms of Mutagenesis, 1988, 197(1): 作者简介 ( 责任编辑 : 白洪健 ) 缪文彬, 工程师, 主要研究方向为化学品风险评估 miuwb@shciq.gov.cn

Microsoft Word 聂雪梅.doc

Microsoft Word 聂雪梅.doc 4 4 Vol. 4 No. 4 2013 8 Journal of Food Safety and Quality Aug., 2013 聂雪梅 *, 李立, 孙利, 高飞 (, 100123) 摘要 : 2011 12,, 2007~2011,,,, 关键词 : ; ; ; ; Trade and notification situation on food import and export