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目的 寻求单细胞凝胶电泳计算机成像与分析系统中的有效分析指标 ,为H2 O2 在 1℃条件下作为检测DNA交联的标准DNA断裂剂提供依据。方法 常规培养Vero细胞 ,消化后悬浮在PBS中 ,H2 O2 染毒 ,分别在 37℃和 1℃条件下孵育 1h ,常规单细胞凝胶电泳 ,专用软件分析其图像。结果 H2 O2 的染毒浓度在 2 5 0 0 μmol/L以下时 ,尾长、尾面积、尾矩和尾积分等指标均有剂量 -效应关系 (T =37℃时 ,r分别为 0 .6 80 ,0 .72 5 ,0 .796 ,0 .798;T =1℃时 ,r分别为 0 .6 85 ,0 .717,0 .6 2 2 ,0 .714,P均 <0 .0 1)。在H2 O2 浓度较高的另一实验中 ,当H2 O2 为 2～ 7μmol/L时 ,仅尾矩和尾积分间存在这种关系 ;H2 O2 各剂量 (2 0、10 0、5 0 0、2 5 0 0 μmol/L)在 37℃时对DNA的损伤程度均明显高于1℃时。两种条件下上述 4个指标都存在剂量 -效应关系。结论 由该系统得到的尾矩和尾积分所反映的DNA损伤信息更准确、更全面、适用范围更广 ,但尾长可用作较初步的研究 ;H2 O2 在 1℃条件下可以作为检测DNA交联的标准DNA断裂剂
OBJECTIVE: To find an effective analytical indicator in the single-cell gel electrophoresis computer imaging and analysis system and to provide a basis for H2O2 as a standard DNA fragmentation reagent for DNA cross-linking at 1 ℃. Methods Vero cells were cultured routinely. After digestion, the cells were suspended in PBS and incubated with H 2 O 2. The cells were incubated at 37 ℃ and 1 ℃ for 1 h, respectively. Conventional single-cell gel electrophoresis and special software were used to analyze the images. Results The tail length, tail area, tail moment and tail integral of H 2 O 2 were all dose-dependent when the concentration was below 2500 micromol / L (r = 0.6 at T = 37 ℃ 80, 0 .72 5, 0 .796, 0 .798; when T = 1 ℃, r were respectively0.685,0.717,0.62 2,0.714, P <0.01 ). In another experiment with high concentration of H2O2, only the tail moment and the tail integral existed when the H2O2 was 2 ~ 7μmol / L; each dose of H2O2 (2 0, 10 0, 5 0 0, 2500μmol / L) at 37 ℃, the DNA damage was significantly higher than at 1 ℃. Under the two conditions, there are dose-effect relationship among the above four indexes. Conclusion The DNA damage information reflected by the tail moment and tail integral obtained by the system is more accurate, more comprehensive and more applicable. However, the tail length can be used as a preliminary study. H2O2 can be used as a detection DNA at 1 ℃ A standard cross-linked DNA breaker