MTT法用于药物对L2细胞毒性的实验研究

目的：评价噻唑蓝（MTT）法检测药物对细胞的毒性作用的可靠性。方法：大鼠肺泡上皮L2细胞以叔丁基对苯二酚（TBHQ）10．100μM，BsO以1-10mM分别处理，用MTT法检测细胞活性、JC-1（5,5’，6，6’-四氯．1，1’，3，3’-四乙基苯并咪唑羰花青碘化物）荧光染料法检测细胞线粒体电位改变、台盼蓝排斥实验检测细胞死亡率，分析各指标的情况。结果：在处理剂量范围，MTT法检测到的光密度（OD）值未能达到一般判断的半数抑制浓度（ic50）水平，最高抑制率仅达到30％左右；台盼蓝排斥试验检测数据表明TBHQ的LC50值为50μM，丁硫氨酸亚砜胺（BSO）为5mM；利用JC-1荧光染料判断的半数凋亡剂量分别为50μM和7mM。结论：MTT法作为最常采用的细胞生长抑制检测手段，但在某些特定实验中可能不能客观地反映细胞的活性，建议多种方法结合进行评价。

MTT Assay for Cytotoxic Drugs on Experimental Study of L2 Cells

Objective： To evaluate the reliability of toxic effects of drugs using MTT assay. Methods：The rat alveolar epithelial L2 cells exposure to TBHQ 10-100μM, BSO 1-10 mM, respectively, using MTT assay to detect the activity of cells, JC-1 fluorescent dye to detect the changes in mitochondrial potential, trypan blue exclusion to detect the prevalence. Results： In the treatment dose range, the OD values detected by MTT assay failed to meet generally determine the IC 50 level, and the highest inhibition rate is around 30%; trypan blue exclusion assay data indicate that the LC 50 value of TBHQ is 50 μM, BSO to 5 mM ; using of JC-1 fluorescent dye to determine the half dose is 50 μM and 7 mM, respectively. Conclusion： MTT assay is the most common means to detect the cell growth inhibition, but it may not objectively reflect the activity of cells in a given experiment. We suggest to use a variety of methods in conjunction.