矢车菊素-3-葡萄糖苷干预丙烯酰胺细胞毒性的体外模型筛选

Screening in Vitro Model for Acrylamide-induced Cytotoxicity Intervention by Cyanidin-3-glucoside

丙烯酰胺是食品热加工过程中形成的一种内源性化学污染物,能引起细胞毒性。矢车菊素-3-葡萄糖苷作为一种果蔬中广泛存在的花色苷,具有显著的抗氧化活性。目前应用花色苷进行AA细胞毒性的干预尚无系统性研究。为了筛选适用于AA细胞毒性干预的细胞模型,对体外培养的Hep G2、L02、Caco-2、BHK-21及MDA-MB-231等细胞,通过不同浓度AA和Cy-3-glu培养,采用结晶紫染色法测定不同时间的细胞存活率,最终确定AA最适的作用时间为24 h,适宜作用浓度分别为2.5 mmol/L和5.0 mmol/L;Cy-3-glu的最适预处理时间为4 h。筛选出适合Cy-3-glu干预的AA诱导的细胞模型为MDA-MB-231细胞。通过Cy-3-glu抑制细胞内活性氧生成和谷胱甘肽的降低并验证,10~100μmol/L Cy-3-glu预处理表现出显著的AA毒性的保护作用,为毒性干预研究提供模型基础。

Acrylamide （AA）, which can cause cytotoxicity, is an endogenous chemical contaminant during food ther- mal processing. Cyanidin-3-glucoside （Cy-3-glu）, as one kind of anthocyanin widely occurring in fruits and vegetables, has obvious antioxidant activity. Recently, there is no systematic study on the intervention against AA cytotoxicity with Cy-3-glu. There were not the systemic studies on the intervention against AA eytotoxicity with Cy-3-glu. The aim of this study is to screen and obtain the suitable cell model to be applied to AA-induced toxicity in vitro. Five cells （HepG2, L02, Caco-2, BHK-21 and MDA-MB-231） were cultured at different AA concentrations and Cy-3-glu. Cell viability was measured using the crystal violet staining and was used as the indicator for cytotoxicity. For AA treatment, the opti- mal incubation duration was 24 h and the suitable concentration was 2.5 mmol/L or 5.0 mmol/L. Moreover, the optimal Cy-3-glu pretreatment time was 4 h. Among five cells, the MDA-MB-231 cells are the most suitable cell model applied for the study on the intervention against AA toxicity with 10-100 μmol/L Cy-3-glu. In addition, Cy-3-glu showed the protective role against reactive oxygen species （ROS） and glutathione （GSH） depletion induced by AA in MDA-MB-231 cells. It would provide a model basis for future research on toxicity intervention.