Wang Xiangfei, Ge Junbo, Wang Keqiang, Qian Juying, Zou Yunzeng
Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, People's Republic of China.
Assay Drug Dev Technol. 2006 Apr;4(2):203-7. doi: 10.1089/adt.2006.4.203.
The specificity and sensitivity of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium assay can be influenced by certain factors, such as colored substances and cell volume. When the MTT assay is applied to measure cytotoxicity induced by emodin, its accuracy might be affected by emodin itself. Vascular smooth muscle cells were cultured in M199 medium. The optical density of emodin or formazan was measured by spectrophotometry. Emodin has a different absorption spectrum in different solvents. The solvents containing water induced a red shift of the absorption curve of emodin, which increased the overlap of the absorption curves of emodin and formazan. Formazan was formed from the MTT tetrazolium salt by emodin in a dose-dependent manner, which was partially suppressed by serum. Cytotoxicity was induced by emodin in a time- and dose-dependent manner in a modified MTT assay. The data suggest that emodin can alter the accuracy of the MTT assay but that a modified MTT assay is still valuable in measuring emodin-induced cytotoxicity.
3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）比色法的特异性和敏感性会受到某些因素的影响，如有色物质和细胞体积。当MTT法用于检测大黄素诱导的细胞毒性时，其准确性可能会受到大黄素本身的影响。血管平滑肌细胞在M199培养基中培养。通过分光光度法测量大黄素或甲臜的光密度。大黄素在不同溶剂中有不同的吸收光谱。含水溶剂会导致大黄素吸收曲线发生红移，这增加了大黄素和甲臜吸收曲线的重叠。大黄素以剂量依赖的方式由MTT四氮唑盐形成甲臜，血清可部分抑制这一过程。在改良的MTT法中，大黄素以时间和剂量依赖的方式诱导细胞毒性。数据表明，大黄素会改变MTT法的准确性，但改良的MTT法在检测大黄素诱导的细胞毒性方面仍然具有价值。
