摘要
目的:探索四甲基偶氮唑盐(methylthiazolyl tetrazolium,MTT)实验方法的最佳条件,并检测脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)的生物学活性。方法:根据MTT法的测定原理,采用三因素(细胞数量、MTT浓度、MTT染色时间)三水平的正交实验设计;并依据MTT的最优条件测定BDNF的生物活性。结果:MTT比色法检测SH-SY5Y细胞活性的最优条件为:每孔细胞数:(0.2×108个/L,100μl),MTT的染色时间(6小时),MTT的剂量(5g/L)。在ATRA诱导SH-SY5Y细胞产生TrkB受体后,随着BDNF浓度的增加,吸光值也增加,当BDNF浓度增加到50μg·L-1后,随着BDNF浓度的增加,吸光值增加并不显著。结论:建立了MTT比色法的最佳条件,并应用于BDNF细胞因子的活性检测中,在BDNF浓度为50μg·L^(-1)时。
Objective：To investigate the optimal conditions for MTT colorimetry and detection of BDNF biological activities.Methods：To use the orthogonal test by taking SH-SY5Y cells as the subject and according to the principle of MTT colorimetry. The orthogonal design adopted three factors： cell number, MTT concentration, dyeing time in three levels. And the biological activities of BDNF were detected under the optimal conditions. Results：The optimal conditions for MTT colorimetry was determined as follows： （0.2x 108/L, 100 μl） as the cell number in each well, 6 hours as the staining time, and 5 g/L as the dose in the MTT. The biological activities of BDNF were detected under these conditions. When the concentration of BDNF was 50 μg-L-1, the cellular absorbance was not significantly increased along with increased concentrations of BDNF. Conclusion：The optimal conditions for MTT colorimetry was determined using the orthogonal test by taking SH-SY5Y cells as the subject and the biological activities of BDNF were detected under these conditions. It was able to get favorable dose-effect relation.
出处
《中国医药导刊》
2011年第5期905-907,共3页
Chinese Journal of Medicinal Guide
基金
军队十一五科技攻关项目(项目编号06G119)
国家重大新药创制项目(项目编号2009ZX0913-722)
关键词
四甲基偶氮唑盐
脑源性神经营养因子
正交实验
Methylthiazolyl tetrazolium
Brain-derived neurotrophic factor
Orthogonal test
