【摘要】 目的研究氨磷汀对噻唑蓝(MTT)实验结果的影响。方法实验共分4组:调零组(培养基)、对照组(培养基和细胞)、氨磷汀组(培养基和氨磷汀)、氨磷汀+细胞组(培养基、细胞和氨磷汀),如常规实验加入MTT,立即观察颜色改变及酶联免疫检测仪490 nm处测吸光度(A)值,孵育4 h后再次检测各组A值。结果①调零组、对照组加MTT后均无颜色改变,A值一直在很低水平(0.087±0.021和0.093±0.018),孵育4 h仍无呈色反应,A值也无明显变化(0.076±0.015和0.103±0.022,P>0.05);②氨磷汀组、氨磷汀+细胞组加MTT后均立即呈现蓝紫色,0.5 mmol.L-1氨磷汀时A值分别为0.215±0.013和0.274±0.013,较调零组、对照组均明显升高(P<0.01),且随着氨磷汀浓度增高A值也明显升高(P<0.01);孵育4 h后氨磷汀组A值无继续升高(0.247±0.031,P>0.05),而氨磷汀+细胞组A值仍有升高(0.381±0.022,P<0.05)。结论氨磷汀本身及代谢产物可能与MTT直接反应并呈色,改变MTT实验中的A值,干扰MTT实验结果。更多还原

【Abstract】 Objective To investigate the impact of amifostine(AMI) on the result of MTT assay. Methods Four groups were arranged as: reset group(medium),control group(medium+cell)、AMI group(AMI+medium) and AMI+Cell group(AMI+medium+cell).At the point of MTT adding and 4 h later,the color of cell culture was observed and the A was detected by enzyme-linked immune assay. Results The results showed that the color of culture was unchanged in both reset group and control group,and the of which were low after adding MTT(0.087±0.021 and 0.093±0.018).Even incubated for 4 hours,the color and A still didn’ t change(0.076±0.015 and 0.103±0.022,P>0.05);the color changed into amethyst in both AMI group and AMI+Cell group,and the A were 0.215±0.013 and 0.274±0.013 after adding MTT with AMI at 0.5 mmol·L-1,which were significantly higher than those in the reset and control group in a dose-dependent way of AMI.After incubating for 4 hours,the A ceased increasing in the AMI group(0.247±0.031,P>0.05),but continued to elevate(0.381±0.022,P<0.05) in the AMI+Cell group. Conclusion It is concluded that AMI and its metabolites may directly react with MTT and change the color and value,impacting the result of MTT assay.更多还原