摘要
目的探讨氟化钠(NaF)对体外大鼠软骨细胞活性及DNA损伤作用的影响。方法自2月龄SPF级雌性SpragueDawley大鼠制取膝关节软骨细胞悬液。取对数生长期细胞(细胞密度为2×105个/孔),用含NaF终浓度为0(阴性对照)、0.1、1.0、10.0、20.0、40.0mmol/L的培养基分别处理2、4、8、24h,同时设立空白对照(DMEM/F12)组,采用MTT法检测细胞活性。根据MTT检测的结果,取对数生长期细胞(细胞密度为2×105个/孔),用0(阴性对照)、0.1、0.5、1.0、2.0、5.0、10.0mmol/LNaF染毒4h,阳性对照组在紫外灯下照射10min。用单细胞凝胶电泳技术(SCGE)检测细胞DNA的损伤情况。结果与阴性对照组比较,染毒2h后20.0、40.0mmol/LNaF染毒组和染毒4、8、24h后10.0、20.0、40.0mmol/LNaF染毒组大鼠软骨细胞的细胞活性均下降,差异有统计学意义(P<0.05或P<0.01)。与阴性对照组相比,各浓度NaF染毒组大鼠软骨细胞DNA损伤率均较高,差异有统计学意义(P<0.01)。且随着NaF染毒浓度的升高,大鼠软骨细胞DNA损伤率呈升高趋势。与阴性对照组相比,1.0～10.0mmol/LNaF染毒组大鼠软骨细胞尾长、Olive尾矩和彗星矩较高,差异有统计学意义(P<0.01)。且随着NaF染毒浓度的升高,大鼠软骨细胞尾长、Olive尾矩和彗星矩均呈上升趋势。结论在体外培养条件下,NaF能明显抑制大鼠软骨细胞的增殖,其作用机制可能与DNA损伤有关。
Objective To investigate the effect of fluoride on cytoactive and DNA damage in chondrocytes of rat in culture. Methods Chondroeytes were separated from the knee joints cartilage of female SD rats, SPF, aged 2-months and cultured, cell density was 2×10^5/hole. Six groups were designed, including the control group and NaF group （0.1, 1.0, 10.0, 20.0 and 40.0 mmol/L）. After 2, 4, 8 and 94 h of administration, the inhibitory effect of NaF on the growth of chondrocytes was evaluated by MTT assay. SCGE was used to detect the DNA damage of chondrocytes after exposure to NaF of different concentration（O, 0.1, 0.5, 1.0, 2.0, 5.0 and 10.0 mmol/L） for 4 hours. The positive control was exposed to UV light for 10 min. Results Compared with the negative control group, NaF of 20.0 and 40.0 mmol/L for 2 h and NaF of 10.0, 20.0 and 40.0 mmol/L for 4, 8 and 24 h could obviously inhibit chondrocytes proliferation （P〈0.05 and P 〈0.01 ）. Compared with the negative control group, DNA damage rate of chondrocytes in all NaF groups increased significantly （P〈0.01）. With the level of NaF increased, all the analysis indexes including tail length, Olive tail moment and comet moment increased in a dose-dependent manner. Conclusion NaF may present an obvious inhibitory effect on viability and growth in chondrocytes in culture, maybe DNA damage is concerned in the mechanism.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2010年第10期875-877,共3页
Journal of Environment and Health
基金
上海市卫生局科研课题局级科研项目(2007180)
