论文部分内容阅读
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 sodium fluoride (NaF) on chondrocyte activity and DNA damage in vitro. Methods Knee chondrocyte suspensions were prepared from 2-month-old SPF female Sprague Dawley rats. The cells in logarithmic growth phase (cell density of 2 × 10 5 cells / well) were treated with medium containing NaF at a final concentration of 0 (negative control), 0.1, 1.0, 10.0, 20.0 and 40.0 mmol / L, , 8,24h respectively. At the same time, a blank control (DMEM / F12) group was set up and the cell viability was measured by MTT assay. According to the results of MTT assay, cells in logarithmic growth phase (cell density 2 × 10 5 cells / well) were treated with 0 (negative control), 0.1, 0.5, 1.0, 2.0, 5.0 and 10.0 mmol / The control group was exposed to ultraviolet light for 10 minutes. Cell DNA damage was detected by single cell gel electrophoresis (SCGE). Results Compared with the negative control group, the cell viability of chondrocytes in 20.0 and 40.0 mmol / L NaF groups and in the groups of 10.0, 20.0 and 40.0 mmol / L NaFF groups after 4, 8 and 24 hours of exposure were both decreased, The difference was statistically significant (P <0.05 or P <0.01). Compared with the negative control group, the DNA damage rate of chondrocytes in NaF group was higher than that in negative control group, the difference was statistically significant (P <0.01). And with the increase of NaF concentration, the DNA damage rate of rat chondrocytes increased. Compared with the negative control group, the tail length, Olive tail moment and comet moment of chondrocytes in 1.0 ~ 10.0 mmol / L NaF treatment group were significantly higher (P <0.01). And with the increase of NaF concentration, tail length, Olive tail moment and comet moment of chondrocytes increased. Conclusion NaF can significantly inhibit the proliferation of rat chondrocytes in vitro under the condition of in vitro culture, and its mechanism may be related to DNA damage.