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目的研究互隔交链孢酚(AOH)对小鼠胚胎成纤维细胞NIH/3T3的急性毒性作用。方法将对数生长期的NIH/3T3细胞分为10、50μmol/L AOH处理组和溶剂对照组,观察染毒细胞形态学变化,四甲基偶氮唑蓝(MTT)法研究细胞存活率,采用彗星实验观察细胞的DNA损伤程度,利用流式细胞术(FCM)检测对细胞周期的影响。结果细胞处理后,出现明显的细胞形态学变化。MTT结果表明,10、50μmol/L AOH处理组细胞抑制率(分别为19.88%,32.47%)显著高于溶剂对照组(P<0.05)。10、50μmol/L AOH组在彗星实验中细胞拖尾率为35.87%和71.83%,尾部DNA含量为(36.18±18.6)和(51.3±21.6),显著高于溶剂对照组的拖尾率(14.78%)和尾部DNA含量(21.29±15.60)(P<0.05);10μmol/L AOH处理组对NIH/3T3细胞周期影响不明显,而50μmol/L AOH处理组引起S期增高和明显的G2/M期阻滞(P<0.05)。结论AOH可诱导NIH/3T3细胞的DNA损伤、抑制细胞增殖,并诱导G2/M期细胞阻滞。
Aim To study the acute toxicity of AOH on mouse embryonic fibroblasts NIH / 3T3. Methods NIH / 3T3 cells in logarithmic growth phase were divided into 10, 50μmol / L AOH treatment group and solvent control group. Morphological changes of the infected cells were observed. MTT assay was used to study cell viability, Using the comet assay to observe the degree of DNA damage in cells, the effect of cell cycle was detected by flow cytometry (FCM). The results of cell treatment, significant changes in cell morphology. The results of MTT assay showed that the cell inhibitory rates of A50 and AOH treatment groups (19.88% and 32.47%, respectively) were significantly higher than those of the solvent control group (P <0.05). In the 50μmol / L AOH group, the tailing rates were 35.87% and 71.83% in the comet assay and the tail DNA contents were (36.18 ± 18.6) and (51.3 ± 21.6), significantly higher than those in the solvent control group (14.78 (P <0.05); 10μmol / L AOH treatment group had no significant effect on the cell cycle of NIH / 3T3 cells, while 50μmol / L AOH treatment group caused an increase of S phase and significant G2 / M Stage arrest (P <0.05). Conclusion AOH can induce DNA damage, inhibit cell proliferation and induce G2 / M cell cycle arrest in NIH / 3T3 cells.