高氟茶浸泡液致V79细胞毒性及DNA损伤的研究  

Studies on the Cytotoxicity and DNA Damage of Tea Soaking Solution Containing High Fluoride on V79 Cell

作　　者：高美伶[1] 范家林[2] 周建华[1,3] 

机构地区：[1]苏州大学放射医学与公共卫生学院,江苏苏州215123 [2]常熟市疾病预防控制中心,江苏常熟215500 [3]江苏省放射医学与防护重点实验室,江苏苏州215123

出　　处：《环境与职业医学》2009年第5期457-460,共4页Journal of Environmental and Occupational Medicine

基　　金：常熟市社会发展基金资助(编号:C2006060)

摘　　要：[目的]探讨含氟茶浸泡液对中国仓鼠肺成纤维细胞(V79细胞)的毒性及DNA损伤的作用。[方法]用氟离子选择电极法测定绿茶A、绿茶B和对照茶浸泡液中氟化物的含量。3种茶浸泡液分别设立5个茶汤浓度组(0.63、1.25、2.50、5.00、10.00mg/mL)对V79细胞进行4h、24h染毒,采用台盼蓝拒染法检测茶浸泡液对V79细胞的毒性。另将上述3种茶浸泡液分别设0.32、0.63和1.25mg/mL3个浓度组,均对V79细胞染毒4h,采用单细胞凝胶电泳技术(SCGE)检测茶浸泡液对V79的细胞DNA损伤作用。同时设立阴性和紫外线对照组。[结果]绿茶A、绿茶B及对照茶浸泡液中氟化物的含量分别为2330、1390、7.53mg/kg。绿茶A、绿茶B的氟含量高于农业行业标准。在台盼蓝拒染实验中,绿茶A和绿茶B浸泡液对V79细胞活性有抑制作用,并具有明显的剂量-效应关系。在作用4h(2.50～10.00mg/mL剂量范围)和24h(0.63～10.00mg/mL)时,绿茶A和绿茶B的细胞存活率明显低于对照茶(P<0.05),且绿茶A的细胞存活率低于绿茶B(P<0.05)。在作用4h时,绿茶A、绿茶B和对照茶对V79细胞的半数致死浓度(IC50)分别为7.58、9.01、11.81mg/mL。在SCGE中,采用几个常用指标[彗星拖尾率、尾长、Olive尾矩(Olivetailmoment,OTM)、尾分布矩(Tailmoment,TM)、尾部DNA含量]观察。绿茶A和绿茶B的各项指标明显高于对照茶(P<0.05),绿茶A的各项指标均高于绿茶B(P<0.05)。而对照茶在各个浓度上与阴性组之间差异无统计学意义。[结论]2种高氟茶浸泡液对V79细胞活性有抑制作用,并呈现一定的剂量-效应关系,绿茶A对V79细胞的毒性最大。绿茶A和绿茶B的浸泡液可引起V79细胞的DNA损伤,其中绿茶A的DNA损伤作用较大,可能与它含氟量最高有关,而对照茶对V79细胞无DNA损伤作用。[ Objective ] The effects of tea soaks containing high fluoride on cytotoxicity and DNA damage in Chinese hamster lung fibreblast cell（ V79cell ）were studied. [ Methods ] The fluoride content of green tea A, green tea B and the control tea soaking solutions were determined by fluorine ion-selective electrode analysis .V79 cells were exposed at the concentrations of 0.63, 1.25, 2.5, 5, and 10 mg/mL of each tea solution for 4 h and 24 h. Trypan blue stain assay was used to detect the cytotoxicity of the three tea solutions on V79 cell. V79 cells were exposed at the concentrations of 0.32, 0.63, 1.25 mg/mL of each tea solutions for 4 h. The single cell gel electrophoresis（ SCGE ）was used to measure the DNA damage of the three tea solutions. [ Results ] The fluoride content of green tea A and B were 2 330 and 1 390 mg/kg respectively, both exceeded the agriculture profession standard, and of the control tea was only 7.5 mg/kg. In Trypan blue stain assay, high fluoride tea solutions could induce viability inhibition with significant dose-effect relationship. The cell survival rate of the green tea A and B were significantly less than the control tea （ P 〈 0.05 ）, whereas the rate of the green tea B was significantly higher than that of the green tea A （ P 〈 0.05 ） at the concentration of 2.50 mg/mL-10.00 mg/mL for 4h and 0.63-10.00 mg/mL for 24 h. Median lethal concentrations（ IC50 ）for V79 cells of green tea A, green tea B and control tea were 7.58, 9.01 and 11.81 mg/mL respectively when exposed for 4 hours. In SCGE assay, observed by general indices（ tail rate, tail length, olive tail moment, tail moment, DNA content in tail ）, the two tea solutions of A and B could damage the DNA proportionally with dosages. The general indices of the green tea A and B were significantly higher than the control tea（ P 〈 0.05 ）, whereas the general indices of the green tea A was significantly higher than that of the green tea B（ P 〈 0.05 ）, There was nosignificant difference between the contro

关 键 词：氟化钾 V79细胞 DNA损伤 单细胞凝胶电泳 

分 类 号：R114[医药卫生—卫生毒理学]