不同抗生素对耻垢分枝杆菌细胞蛋白质差异表达  

Detection of protein expression from Mycobacterium smegmatis mc^2155 treated with three drugs

作　　者：湛垚垚[1] 刘晶华 张翠丽[1] 辛毅[1] 

机构地区：[1]大连医科大学生物技术教研室,大连116027 [2]大连市结核病医院

出　　处：《中国公共卫生》2008年第5期553-555,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金项目(30570416)

摘　　要：目的利用蛋白质组学方法,识别乙胺丁醇(EMB)、异烟肼(I NH)、利福平(RIF)处理前后耻垢分枝杆菌mc2155细胞差异表达的蛋白质,探讨3种抗生素抑制分枝杆菌生长的作用机制及寻找新的抗结核药物作用靶标。方法采用载体两性电解质pH梯度双向凝胶电泳技术分离乙胺丁醇、异烟肼、利福平处理前后耻垢分枝杆菌mc2155细胞的总蛋白质,用PDQuest 7.3.1图像分析软件比较分析,以识别比较差异表达的蛋白质。结果不同浓度的乙胺丁醇、异烟肼和利福平对耻垢分枝杆菌细胞的增殖均有不同程度的抑制作用。处理3 h时,耻垢分枝杆菌mc2155细胞增殖首次受到明显抑制;3种抗生素最佳处理浓度分别为:乙胺丁醇24.0μg/ml,异烟肼0.08μg/ml,利福平0.8μg/ml。比较蛋白质组分析显示,对照组有155个蛋白斑点,乙胺丁醇处理组有蛋白斑点147个,异烟肼处理组有蛋白斑点235个,利福平处理组有蛋白斑点161个。对照组、乙胺丁醇处理组,异烟肼处理组和利福平处理组的蛋白斑点的分布明显不同。结论3种抗生素处理前后,耻垢分枝杆菌mc2155细胞蛋白质组的差异分析有助于进一步研究抗生素的作用机制,并为寻找新的抗结核药物作用靶标提供基础依据。Objective To investigate the microbiostatic mechanism of ethambutol （EMB）, isoniazid （I NH） and rifampin （RIF）, and to find differential expression proteins in Mycobacterium smegmatis mc2155 cells treated by ethambutol, isoniazid and rifarnpin, respectively. Methods The total protein of untreated and etharnbutol -, isoniazid - and rifampin - treated mc2155 cells were separated by ampholyte pH gradients based on two- dimensional gel electrophoresis（2 - DE）. The differentially expressed proteins were analyzed by using image analysis software. Results Control group and ethambutol -, isoniazid - and riampin - treated group contained 155,147,235 and 161 protein spots, respectively. Distributions of proteins of control group and ethambutol -, isoniazid - and rifampin - treated group have different pattern. Conclusion There is a significant difference in protein expression between untreated and ethambutol- , isoniazid- and rifampin - treated mc2155 cells. It suggests that the differential expression analysis of proteomes may be useful to further elucidate microbiostatic mechanism of ethambutol, isoniazid and rifampin, and search for new drug targets against TB.

关 键 词：乙胺丁醇 异烟肼 利福平 耻垢分枝杆菌mc2155 双向电泳 蛋白质组比较 

分 类 号：R978.1[医药卫生—药品]