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目的探究双酚A(bisphenol A,BPA)对Caco-2细胞的毒性及氧化应激作用。方法选用6.25、12.5、25、50和100 mg/L的BPA与Caco-2细胞共培养24 h,MTT法和乳酸脱氢酶(LDH)释放法测定BPA对Caco-2细胞毒性作用,比色法检测细胞中抗氧化防御酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)以及脂质过氧化产物丙二醛(MDA)的活力和含量,荧光探针法检测活性氧簇(ROS)生成。结果 50、100 mg/L BPA与Caco-2细胞共培养24 h后,细胞存活率与对照组比较显著降低(P<0.01);细胞抗氧化防御酶SOD、CAT活力下降,MDA含量增加,与BPA浓度呈正相关;50、100 mg/L BPA处理组,细胞ROS生成量与对照组比较显著升高(P<0.01),ROS荧光信号与BPA浓度呈正相关。结论结果表明BPA对Caco-2细胞的毒性作用与其浓度呈正相关,BPA引起Caco-2细胞的氧化应激与其发挥细胞毒性有相关性。
Objective To investigate the toxicity and oxidative stress of bisphenol A (BPA) on Caco-2 cells. Methods Caco-2 cells were cocultured with BPA at 6.25, 12.5, 25, 50 and 100 mg / L for 24 h. The cytotoxicity of BPA on Caco-2 cells was determined by MTT and lactate dehydrogenase (LDH) (SOD), catalase (CAT), and malondialdehyde (MDA), a lipid peroxidation product, were measured by flow cytometry. Fluorescence probe method was used to detect the activity of reactive oxygen species ROS) generation. Results Compared with the control group, the cell survival rate of 50,100 mg / L BPA and Caco-2 cells were significantly decreased (P <0.01), the activities of antioxidant enzymes SOD and CAT decreased and the content of MDA increased (P <0.01). There was a positive correlation between ROS fluorescence signal and BPA concentration in 50,100 mg / L BPA treatment group. Conclusions The results showed that the toxic effect of BPA on Caco-2 cells was positively correlated with the concentration of BPA. The oxidative stress induced by BPA in Caco-2 cells was related to its cytotoxicity.