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目的 研究丙烯酰胺 (AA)对人角质形成细胞的细胞毒性和对DNA的损伤及其可能机制。方法 (1 )人角质形成细胞株HaCaT细胞经AA浓度梯度染毒 44h后 ,以四氮唑盐 (MTT)法检测细胞存活率。 (2 )细胞经AA浓度梯度染毒 ,以彗星试验检测细胞DNA损伤。 (3)细胞经 2 .0 0mmol/LAA和 0 .50mmol/L的细胞色素P 450 (CYP 450 )抑制剂 1 ABT共处理 4h后 ,以彗星试验探索DNA损伤与CYP 450的关系。结果 (1 )细胞毒性试验显示 ,AA染毒 44h后细胞存活率显著下降。 (2 )AA染毒 4h未观察到细胞毒性 ,但各组细胞的DNA较对照组均检测到显著损伤 ,且损伤程度随染毒剂量增大而加重 ,彗尾长度在处理剂量范围内有明显的剂量效应关系 ;细胞经 1 ABT和 2 .0 0mmol/LAA共同处理后 ,拖尾率和尾长为 1 5 .4 %和 (8.2± 2 .0 ) μm ,较 2 .0 0mmol/LAA组 [80 .6 %和 (44.3± 4 .0 ) μm]明显降低 ,差异有显著性 (P <0 .0 1 )。结论 AA对HaCaT细胞具有细胞毒性和基因毒性 ,AA诱导的DNA损伤可能与其经CYP 450作用产生的氧化代谢物有关
Aim To investigate the cytotoxicity of acrylamide (AA) on human keratinocytes and its damage to DNA and its possible mechanism. Methods (1) Human keratinocyte line HaCaT cells were treated with AA concentration gradient for 44 h. Cell viability was detected by MTT assay. (2) Cells were exposed to a gradient of AA concentration and DNA damage was detected by comet assay. (3) The cells were treated with 2.0 mmol / LAA and 0.50 mmol / L cytochrome P450 (CYP450) inhibitor 1 ABT for 4 h, and the relationship between DNA damage and CYP 450 was explored by comet assay. Results (1) Cytotoxicity test showed that cell viability was significantly decreased after AA exposure for 44h. (2) No cytotoxicity was observed after 4 hours of AA exposure, but DNA of each group was significantly damaged as compared with that of the control group, and the degree of injury was aggravated with the increase of the dose, and the length of the tail was significantly The tailing rate and tail length of the cells after treatment with 1 ABT and 2.0 mmol / LAA were 15.4% and 8.2 ± 2.0 μm, respectively, which were significantly higher than those of 2.0 mmol / LAA group [80 .6% and (44.3 ± 4 .0) μm] were significantly lower, the difference was significant (P <0.01). Conclusion AA is cytotoxic and genotoxic to HaCaT cells. AA-induced DNA damage may be related to the oxidative metabolites produced by CYP 450