Yamashoji S, Nishimoto F, Usuda M, Kubota H, Isshiki K
Institute of King Brewing, Kakogawa, Japan.
Anal Biochem. 1992 Dec;207(2):255-60. doi: 10.1016/0003-2697(92)90009-v.
Menadione-catalyzed H2O2 production by viable cells is proportional to viable cell number. The correlations between the viable cell number and the concentration of H2O2 produced are determined with the rapid chemiluminescent assay (S. Yamashoji, T. Ikeda, and K. Yamashoji, 1989, Anal. Biochem. 181, 149-152). This chemiluminescent assay of viable cells requires only 10 min and is much faster than NR (neutral red) inclusion and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction assays, which require 3-5 h. When viable cells are incubated with antitumor drugs, detergents, mycotoxins, and glycoalkaloids for 24-48 h, a decrease in menadione-catalyzed H2O2 production in a dose- or incubation time-dependent manner is observed. In general, the 50% inhibition concentration determined by the chemiluminescent assay is lower than that determined by NR inclusion and MTT reduction assays, and the order of relative cytotoxic effects of agents is the same among these assays. Furthermore, clear cytotoxic effects are observed by the chemiluminescent assay after 1 h exposure of trypsinized cells to toxic compounds. Therefore, the chemiluminescent assay is expected to be more useful for the rapid detection of cytotoxic compounds than NR inclusion and MTT reduction assays.
甲萘醌催化活细胞产生过氧化氢的量与活细胞数量成正比。活细胞数量与所产生的过氧化氢浓度之间的相关性通过快速化学发光测定法确定（S. Yamashoji、T. Ikeda和K. Yamashoji，1989年，《分析生物化学》181卷，第149 - 152页）。这种活细胞的化学发光测定法仅需10分钟，比中性红摄取法和MTT（3 -（4, ,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐）还原法快得多，后两种方法需要3 - 5小时。当活细胞与抗肿瘤药物、去污剂、霉菌毒素和糖苷生物碱一起孵育24 - 48小时时，观察到甲萘醌催化产生的过氧化氢量以剂量或孵育时间依赖性方式减少。一般来说，通过化学发光测定法确定的50%抑制浓度低于通过中性红摄取法和MTT还原法确定的浓度，并且在这些测定中，试剂相对细胞毒性作用的顺序是相同的。此外，在胰蛋白酶消化的细胞暴露于有毒化合物1小时后，通过化学发光测定法可观察到明显的细胞毒性作用。因此，与中性红摄取法和MTT还原法相比，化学发光测定法有望更有助于快速检测细胞毒性化合物。
