7 1 Vol. 7 No. 1 2016 1 Journal of Food Safety and Quality Jan., 2016 缪文彬 *, 蒋伟, 陈相 (, 200135) 摘要 : 目的, 方法 28, 结果 0.5556 mg/ml 1.6667 mg/ml hprt, 5.000 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 200135, 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 0.5556 mg/ml and 1.6667 mg/ml fluorescent brightener 28 treated groups. The hprt gene mutation frequency and micronucleus rate of CHO cells with 5.000 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 :,, E-mail: miuwb@shciq.gov.cn *Corresponding author: MIAO Wen-Bin, Engineer, Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China. E-mail: miuwb@shciq.gov.cn
1, : 189 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 487 476 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, 28 5000 g/ml, 60%, 5 mg/ml,, 5.000 1.6667 0.5556 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),, 0.075 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
190 7,,, 100 mm 200, 3, 37 10 d, Giemsa, 9 d, 6- (6-thioguanine, 6-TG),,,, 6-TG, 3, 37 12 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=0.05 3 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) (%) 0 100 0.5556 mg/ml 0.5556 119.0±1.0 1.6667 g/ml 1.6667 82.4±9.0 5.000 g/ml 5.000 71.8±12.0 90.8±14.9 3.2 荧光增白剂致体外哺乳动物细胞微核数比较 (P<0.01),, 0.5556 m/ml 1.6667 mg/ml, 5.000 mg/ml (P<0.05), 2 5.000 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) ( ) 0 4.40±0.67 0.5556 mg/ml 0.5556 5.99±1.43 1.6667 mg/ml 1.6667 4.00±0.00 5.000 mg/ml 5.000 8.46±0.77 * 31.44±0.62 ** :, * P<0.05, ** P<0.01 3.3 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, 0.5556 mg/ml 1.6667 mg/ml hprt, 5.000 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±9.9 6.9±4.6 0.5556 mg/ml 72.8±7.6 8.6±2.5 0.932 1.6667 mg/ml 46.2±31.8 13.2±7.6 0.116 5.000 mg/ml 75.0±12.9 14.7±4.0 0.039 (5 g/ml) 60.0±5.0 20.7±4.5 <0.001 : * P 4 讨论,, DNA P *
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