活性氧在姜黄素对人胚肾293细胞细胞毒性中的作用

Involvement of reactive oxygen species in curcumin-induced cytotoxicity in 293 cells

目的探讨活性氧在姜黄素所致人胚肾293细胞细胞毒性中的作用。方法姜黄素(0、10、20、30和40μg/ml)处理293细胞24、48和72h后,噻唑蓝(MTT)法观察细胞毒作用;聚乙二醇-超氧化物歧化酶(5U/ml)和聚乙二醇-过氧化氢酶(50U/ml)分别预处理1h,再加入不同浓度的姜黄素(0、10、20、30和40μg/ml),48h后MTT法观察其对姜黄素细胞毒作用的影响;2',7'-二氢二氯荧光比色法测定细胞内活性氧(ROS)水平。结果姜黄素对293细胞的生长抑制呈明显的剂量和时间依赖关系,24、48和72h的IC50值分别为:(24.67±2.24)、(21.96±0.80)和(18.62±0.61)μg/ml;聚乙二醇-超氧化物歧化酶对姜黄素细胞毒作用有明显抑制作用(P<0.05),48h的IC50值为(38.98±1.23)μg/ml,比单独姜黄素处理组的IC50值(21.96±0.80)μg/ml显著升高(P<0.05);10μg/ml以上浓度姜黄素作用293细胞1h即可引起ROS增加(P<0.05或P<0.01)。结论姜黄素能引起293细胞ROS增加,其中超氧化物阴离子在姜黄素的细胞毒作用中起着重要作用。

Objective To investigate the involvement of reactive oxygen species （ROS） in curcumin-induced cytotoxicity in human embryo kidney 293 cells. Methods After 293 cells were exposed to curcumin for 24 h, 48 h and 72 h, MTT assay was performed to assess the cytotoxicity. After pretreatment with superoxide anions cavengers, polyethylene glycol-superoxide dismutase （PEG-SOD）（5 U/ml）or PEG-catalase （50 U/ml）for 1 h, 293 cells were exposed to curcumin for 48 h and MTT assay was performed. The production of reactive oxygen species （ROS） was measured using the 2, 7-dichlorofluorescein diacetate （DCFH-DA） method. Results Exposure of 293 cells to curcumin led to a dose- and time-dependent cytotoxicity with IC50 values of （24.67 ± 2.24）, （21.96 ± 0.80） and （18.62± 0.61 ） μg/ml for 24, 48 and 72 h, respectively. When the cells were pretreated with PEG-SOD, cytotoxicity decreased significantly with IC50 value of （38.98 ± 1.23） μg/ml for 48 h compared to only curcumin-treated cells （ P 〈 0.05）, but PEG- catalase had no effect. The levels of ROS were significantly increased （ P 〈 0.05 or P 〈 0.01 ） when 293 cells were treated with curcumin of concentrations above 10 μg/ml. Conclusion These data suggest that curcumin has a strong effect on ROS production and superoxide anions are critically involved in its cytotoxicity in 293 cells.