二烯丙基硫醚拮抗微囊藻毒素-LR致细胞凋亡

Antagonistic effect of diallyl sulfide on microcystin-LR-induced cell apoptosis

目的探讨二烯丙基硫醚(DAS)对微囊藻毒素-LR(MCLR)诱导的细胞凋亡及活性氧产生的影响。方法体外培养人肝癌细胞株HT17,用1μmol/L MCLR染毒的同时给予DAS(0.02～2 mmol/L)作用4、24h,同时设立溶剂对照组、DAS对照组和单纯MCLR染毒组。应用四甲基偶氮噻唑蓝法检测细胞存活率,流式细胞术检测细胞凋亡,采用荧光探针2-[6-(4'-羟基)苯氧基-3H-占吨醇-3-on-9-yl]-苯甲酸检测细胞内活性氧水平。结果 0.2和2 mmol/LDAS干预组细胞存活率分别为(80.91±5.25)%,(90.13±4.47)%,高于MCLR染毒组的(69.50±3.03)%,差异有统计学意义(P<0.05);而0.2和2 mmol/L DAS干预组细胞凋亡率分别为(10.87±0.52)%,(8.52±0.55)%,低于MCLR染毒组的(13.29±1.16)%,差异有统计学意义(P<0.05);0.2和2 mmol/L DAS干预组的活性氧水平明显下降(P<0.05)。结论 DAS可拮抗MCLR诱导的活性氧产生及细胞凋亡。

Objective To study the effects of diallyl sulfide（DAS） on apoptosis in cells and reactive oxygen species（ROS） generation induced by microcystin-LR（MCLR）.Methods Human hepatoma cell line HT17 were treated with solvent alone or 1μmol/L MCLR in the presence or absence of DAS（0.02-2 mmol/L） for 4 hr or 24 hr.Then the cell viability was measured by methyl thiazolyl tetrazolium（MTT） assay,and the generation of ROS was determined by fluorescent probe 2-[6-（4′-hydroxy）phenoxy-3H-xanthen-3-on-9-yl] benzoic acid.Results The cell viabilities in 0.2 and 2 mmol/L DAS intervention group were 80.91±5.25% and 90.13±4.47%,significantly higher than that of MCLR-treated group（69.50±3.03%,P0.05）.The percentages of apoptosis in 0.2 and 2 mmol/L DAS intervention group were 10.87±0.52% and 8.52±0.55%,significantly lower than that of MCLR-treated group（13.29±1.16%,P0.05）.The levels of ROS in 0.2 and 2 mmol/L DAS intervention group decreased significantly（P0.05）.Conclusion The results suggest that DAS inhibits both ROS generation and cell apoptosis induced by MCLR in HT17 cells.