Cytotoxicity and apoptosis induction by butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT).
Cytotoxicity and apoptosis induction by butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT).
复制标题
丁基羟基苯甲醚 (BHA) 和丁基羟基甲苯 (BHT) 的细胞毒性和细胞凋亡诱导。
作者：
M. Saito;H. Sakagami;S. Fujisawa
以人早幼粒白血病细胞株HL-60和人鳞状细胞癌细胞株HSC-2为实验材料，研究了叔丁基羟基茴香醚（BHA）、叔丁基羟基甲苯（BHT）、2-叔丁基-4-甲基苯酚（BMP）及其混合物（BHA/BHT）（1：1，摩尔比）的细胞毒性和诱导凋亡活性。50%细胞毒性浓度（CC 50）按BHA、BHT（0.2-0.3 mM）> BHA/BHT（0.04-0.07 mM）> BMP（0.02-0.05 mM）的顺序下降。抗氧化剂（N-乙酰-L-半胱氨酸、抗坏血酸钠、过氧化氢酶）的加入降低了BHA/BHT或BMP对HSC-2细胞的细胞毒性，但对BHA或BHT的细胞毒性无影响，而添加NADH（一种醌还原酶）则增强了BMP的细胞毒性。这些结果表明，BHA/BHT和BMP的细胞毒性可能是由活性中间体引起的。BHA诱导的细胞毒性增强辣根过氧化物酶，这表明BHA是可氧化的，并产生细胞毒性BHA自由基。BHA/BHT和BMP较好地诱导HL-60细胞核小体间DNA断裂，BHA次之。BHA/BHT或BMP对HL-60细胞MnSOD mRNA表达有明显的抑制作用，并伴有MnSOD在聚丙烯酰胺凝胶上电泳迁移率的改变。这些化合物在HL-60细胞中激活caspase-3、8和9。半胱天冬酶，特别是半胱天冬酶-3的活化，以BHA/BHT > BHA > BMP > BHT的顺序下降。最具细胞毒性的BMP在最小程度上激活了caspase-3活性，这可能部分是由于坏死的发生。BHA/BHT具有很强的细胞毒性和细胞凋亡诱导作用，这可能是由于BHA苯氧基自由基与BHT或BHT苯氧基自由基相互作用产生的反应性中间体。
The cytotoxicity and apoptosis-inducing activity of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and 2-tert-butyl-4-methylphenol (BMP) and the mixture of BHA and BHT (BHA/BHT) (1:1, molar ratio) were investigated, using human promeylocytic leukemia cell lines (HL-60) and human squamous cell carcinoma cell lines (HSC-2). The 50% cytotoxic concentration (CC50) declined in the order of BHA, BHT (0.2-0.3 mM) > BHA/BHT (0.04-0.07 mM) > BMP (0.02-0.05 mM). The addition of antioxidants (N-acetyl-Lcysteine, sodium ascorbate, catalase) reduced the cytotoxicity of BHA/BHT or BMP against HSC-2 cells, but not that of BHA or BHT, whereas the addition of NADH, a quinone reductase to BMP, enhanced the cytotoxicity. These findings suggested that the cytotoxicity of BHA/BHT and BMP might be caused by reactive intermediates. BHA-induced cytotoxicity was enhanced by horseradish peroxidases, suggesting that BHA was oxidizable and produced cytotoxic BHA radicals. Internucleosomal DNA fragmentation of HL-60 cells was preferably induced by BHA/BHT and BMP, followed by BHA. The MnSOD mRNA expression in HL-60 cells assayed by reverse transcriptase-polymerase chain reaction was highly inhibited by BHA/BHT or BMP, accompanied by the change in the electrophoretic mobility of MnSOD on polyacryamide gel. These compounds activated caspase-3, 8 and 9 in HL-60 cells. Activations of caspases, particularly caspase-3, declined in the order of BHA/BHT > BHA > BMP > BHT. The most cytotoxic BMP activated caspase-3 activity to the least extent, possibly in part due to the occurrence of necrosis. The great cytotoxicity and apoptosis induction by BHA/BHT may be due to reactive intermediates derived from the interaction between BHA phenoxyl radical and BHT or BHT phenoxyl radical.