多孔板-MTT比色法测定植物抗菌成分对细菌的抑制活性  

Detection of Plant Antibacterial Components by Using Microplate-MTT Colorimetric Assay

作　　者：唐静[1] 谈满良[1] 赵江林[1] 汪冶[2] 周立刚[1] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193 [2]贵州省中国科学院天然产物化学重点实验室,贵阳550002

出　　处：《天然产物研究与开发》2008年第6期949-952,共4页Natural Product Research and Development

基　　金：高等学校博士学科点专项科研基金(20060019048);教育部新世纪优秀人才支持计划(NCET-05-0134);国家自然科学基金(30871662)

摘　　要：多孔板-MTT比色法测定植物抗菌成分对细菌抑制活性的步骤为:每孔加入浓度为106cfu/mL的供试菌液90μL,然后加入不同浓度的药液10μL,28℃暗培养24 h后,每孔中加入5 mg/mL的MTT溶液10μL,继续培养4 h后加入10%二甲基亚砜100μL,振荡30 min,在570 nm处测定溶液的吸光值。采用以上方法,测定植物抗菌成分蓝桉醇对辣椒斑点病黄单胞菌(Xanthomonas vesicatoria)和枯草芽孢杆菌(Bacillus subtilis)的半抑制浓度(IC50)分别为0.158和0.395 mg/mL,小檗碱对溶血葡萄球菌(Staphylococcus haemolyticus)的IC50为0.587mg/mL。结果表明,多孔板-MTT比色法能快速、微量地测定植物成分对细菌的抑制活性。An optimal experimental procedure with microplate-MTT colorimetric assay was obtained ,that was ： 1 ） addition of 90 μL of bacterial suspension in concentration of 106 cfu/mL in each well of a 96-well micro plate ;2） then addition of 10 μL of the component solution with different concentration ;3 ） cultivation for 24 h at 28 0℃ in darkness ;4） then 10 μL of MTT solution in 5 mg/mL was added in each well;5） after incubation for another 4 h at 28 ℃ ,100 μL of 10% DMSO was added;6） after shaking for another 30 min ,light absorption values were obtained at 570 nm for the snpernatant with a micro plate spectrophotometer. As the optimal MTT assay mentioned above, globulol from Eucalyptus globulus fruits showed strong inhibitory activities against Xanthomonas vesicatoria and Bacillus subtilis that median effective inhib- itory concentration（ IC50 ） were 0. 158 and 0. 395 mg/mL respectively. Berberine from Phellodendron chinertse fruits had its strong inhibitory activity against Staphylococcus haemolyticus obtaining 0. 587 mg/mL of IC50. This indicated that plant antibacterial components could be evaluated fast and micro-quantitatively by using microplate-MTT colorimetric assay.

关 键 词：多孔板-MTT比色法 抗菌活性成分 抗细菌活性 

分 类 号：Q946.887[生物学—植物学] R285[医药卫生—中药学]