Yumita Nagahiko, Sakata Isao, Nakajima Susumu, Umemura Shin-ichiro
School of Pharmaceutical Sciences, Toho University, 2-2-1 Miyama, Funabashi, Chiba 274-8510, Japan.
Biochim Biophys Acta. 2003 Mar 17;1620(1-3):179-84. doi: 10.1016/s0304-4165(02)00530-5.
Ultrasonically induced cell damage and active oxygen generation with 4-formyloximeetylidene-3-hydroxyl-2-vinyl-deuterio-porphynyl(IX)-6-7-diaspartic acid (ATX-S10) were compared in the same in vitro insonation setup. Sarcoma 180 cells suspended in air-saturated PBS were exposed to ultrasound at 2 MHz for up to 60 s in the presence and absence of ATX-S10. The viability was determined by Trypan blue exclusion test. Ultrasonically induced active oxygen generation in the presence and absence of ATX-S10 in air-saturated aqueous solutions of 50 mM 2,2,6,6-tetramethyl-4-piperidone was detected by electron spin resonance (ESR). Significant enhancement of the rates of both ultrasonically induced cell damage and nitroxide generation was demonstrated with 40-160 microM ATX-S10. Both rates correlated very well. The enhancement of both rates with ATX-S10 was suppressed by 10 mM histidine. These results suggest that ultrasonically generated active oxygen plays a primary role in the ultrasonically induced cell damage in the presence of ATX-S10.
在相同的体外超声照射装置中，比较了4-甲酰氧基亚甲基-3-羟基-2-乙烯基-氘代卟啉基（IX）-6-7-二天冬氨酸（ATX-S10）超声诱导的细胞损伤和活性氧生成情况。将悬浮于空气饱和PBS中的肉瘤180细胞，在有和没有ATX-S10存在的情况下，于2 MHz频率下暴露于超声长达60秒。通过台盼蓝排斥试验测定细胞活力。通过电子自旋共振（ESR）检测在50 mM 2,2,6,6-四甲基-4-哌啶酮的空气饱和水溶液中，有和没有ATX-S10时超声诱导的活性氧生成情况。结果表明，40 - 160 microM的ATX-S10能显著提高超声诱导的细胞损伤率和氮氧化物生成率。这两种速率具有很好的相关性。10 mM组氨酸可抑制ATX-S10对这两种速率的提高作用。这些结果表明，在ATX-S10存在的情况下，超声产生的活性氧在超声诱导的细胞损伤中起主要作用。
