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目的探讨纳米羟基磷灰石(HA-NPs)对3种正常组织细胞的DNA损伤作用,为HA-NPs毒性机制提供实验依据。方法以正常人肝细胞(L02)、肺支气管上皮细胞(HBE)和小鼠成骨细胞(MC3T3)为研究对象,以不同浓度HA-NPs染毒细胞24 h,采用MTT比色法检测细胞毒性,同时用彗星实验和荧光探针法分别检测HA-NPs对细胞的DNA损伤和活性氧水平。结果 HA-NPs在较低浓度(0~400μg/ml)时促进3种细胞的生长,在大于400μg/ml时则抑制细胞生长。随着HA-NPs浓度的增高,3种细胞的拖尾细胞数、尾部DNA百分比、Olive尾距(OTM)均逐渐增加,与对照组比较差异有统计学意义(P<0.05)。3种细胞的活性氧荧光强度均随HA-NPs浓度增加而增加,相较于对照组差异有统计学意义(P<0.05)。结论 HA-NPs可引起3种受试细胞DNA损伤,其机制可能与HA-NPs诱导细胞产生活性氧有关。
Objective To investigate DNA damage of HA-NPs to three kinds of normal tissue cells and to provide experimental evidence for the toxicity mechanism of HA-NPs. Methods Normal human hepatocytes (L02), lung bronchial epithelial cells (HBE) and mouse osteoblasts (MC3T3) were used as experimental subjects. The cells were exposed to different concentrations of HA-NPs for 24 hours. The cytotoxicity The DNA damage and reactive oxygen species of HA-NPs were detected by comet assay and fluorescent probe respectively. Results HA-NPs promoted the growth of three kinds of cells at the lower concentration (0-400μg / ml), but inhibited the growth of cells at more than 400μg / ml. With the increase of HA-NPs concentration, the number of tail cells, tail DNA and Olive tail (OTM) of 3 kinds of cells increased gradually, which had statistical significance compared with the control group (P <0.05). The fluorescence intensity of reactive oxygen species of three kinds of cells increased with the increase of HA-NPs concentration, which was significantly different from the control group (P <0.05). Conclusion HA-NPs can cause DNA damage in three kinds of cells, which may be related to the induction of reactive oxygen species by HA-NPs.