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。
Objective To study the cytotoxicity of ahernariol (AOH) on NIH/3T3 cells. Methods Logarithmic growth phase NIH/3T3 cells were treated at the dose of 10μmol/L and 50#mol/L AOH as treatment groups and treated with DMSO(0.25% ). The effects of AOH on cell proliferations were assessed by morphologic observasion. Inhibition rates of AOH were determined by MTT assay. Comet assay was used to examine DNA damage induced by AOH. Cell cycle distributions were detected by flow cytometric assay (FCM). Results The cells treated with AOH oecured morlogic changes. The inhibition rates of 10μmol/L and 50tsmol/L AOH were 19.88% and 32.47% respectively. In the comet assay two treatment groups percentages of tailed cells were 35.87% and 71.83% respectively. The DNA contents of the tail were (36.18 ± 18.6) and (51.3 ±21.6) respectively. These datas were more higher than those of the solvent control ( P 〈 0.05). In comparison with the control group, the percents of G2/M and S phase cells were increased after treatment of 50.0μmol/L AOH for 24h ( P 〈 0.05 ). Conclusion AOH could have acute cytotoxic effects on NIH/3T3 cells and inhibite cell proliferation and cause DNA damage. High dose of AOH could induce cell cycle arrest in G2/M phase.