氟化钠的体外遗传毒性
In vitro Genotoxicity of Sodium Fluoride
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摘要:
目的 研究氟化钠对小鼠淋巴瘤细胞(L5178Y TK+/-3.7.2c)微核的影响及其对中国仓鼠肺成纤维细胞(CHL)细胞染色体畸变及细胞凋亡的影响。 方法 以灭菌注射用水为阴性对照,采用胞质分裂阻滞微核细胞组学试验法,并以丝裂霉素(0.1μg/mL)为阳性对照,研究氟化钠3个浓度组(100、75与50μg/mL)对L5178Y TK+/-3.7.2c细胞总微核率、核分裂指数、核质桥发生率、核芽突发生率与含核突芽的双核细胞数/含Ⅰ型微核与含Ⅱ型微核的双核细胞数值的影响。以空白细胞为阴性对照,丝裂霉素(0.2μg/mL)为阳性对照,氟化钠以600、360、216和130μg/mL的浓度作用于CHL细胞,以观察其对CHL细胞致突变的作用。用流式细胞仪AnnexinV/PI双染法检测氟化钠对CHL细胞凋亡的影响。 结果 与阴性对照组的相比:①阳性对照组(丝裂霉素,0.1μg/mL)和75、100μg/mL氟化钠组L5178Y TK+/-3.7.2c细胞微核率明显升高(P < 0.05),且氟化钠三个浓度的氟化钠染毒组细胞核质桥发生率、含核突芽的双核细胞数/含Ⅰ型微核与含Ⅱ型微核的双核细胞数值均升高(P < 0.01);②阳性对照组(丝裂霉素,0.2μg/mL)CHL细胞畸变率明显升高(P < 0.05);氟化钠各染毒组畸变率差异无统计学意义(P > 0.05),其早期凋亡率随染毒浓度的增加而上升(r=0.814,P < 0.05)。 结论 氟化钠对L5178Y TK+/-3.7.2c细胞染色体具有损伤作用,对CHL细胞没有致突变作用,可以诱导CHL细胞的早期凋亡,呈现剂量相关性。 Abstract:Objective To assess the effects of sodium fluoride (NaF) on mouse lymphoma cells (L5178Y TK+/-3.7.2c) micronucleus and on chromosome aberration and apoptosis of China hamster lung fibroblast cells (CHL). Methods L5178Y TK+/-3.7.2c cells were treated with sterile water for injection (negative control), mitomycin (0.1 μg/mL) (positive control), and NaF (100, 75, and 50 μg/mL) to evaluate frequency of micronuclei, nuclei division index, nucleoplasmic bridges rate, nuclei buds rate, and nucleoplasmic bridges/micronuclei ratio. CHL cells were treated with blank (negative control), mitomycin (0.2 μg/mL) (positive control), and NaF (130, 216, 360, and 600 μg/mL) to evaluate selected mutagenic effects. AnnexinV/PI double staining method was applied for detecting cell apoptosis induced by NaF. Results Compared with the negative control group:① For L5178Y TK+/-3.7.2c cells, the positive control group treated with 0.1 μg/mL mitomycin and the NaF groups treated with 75 μg/mL and 100 μg/mL had higher frequencies of micronuclei (P < 0.05), and the three NaF groups showed elevated nucleoplasmic bridge rates and nucleoplasmic bridge/micronuclei (P < 0.01); ② For CHL cells, the positive control group treated with 0.2 μg/mL mitomycin had statistical differences in chromosome aberration (P < 0.05); no significant difference of chromosome aberration was observed in the CHL cells exposed to various NaF concentrations (P > 0.05), and the early apoptosis rate rose with elevated concentrations of NaF (r=0.814, P < 0.05). Conclusion NaF could damage L5178Y TK+/-3.7.2c cell chromosome. NaF shows no mutagenic effect on CHL cells, but could induce CHL cell apoptosis in a dose-dependent manner.
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