抗痨颗粒对耐多药结核分枝杆菌蛋白质组学的影响  

Effect of Anti-tuberculosis Particles on Proteomics of Drug-resistant Mycobacterium Tuberculosis

作　　者：李建国[1] 刘湘花[1] 汤红琴[1] 董修兵[1] 李宁宁[1] 

机构地区：[1]河南中医学院病理实验中心,郑州450008

出　　处：《中国实验方剂学杂志》2012年第15期205-210,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省重点科研项目9404758

摘　　要：目的:应用蛋白质组学的双向电泳技术,比较分析抗痨颗粒提取物作用前后耐多药结核分枝杆菌的全菌蛋白表达差异,揭示药物作用机制。方法:临床耐多药结核分枝杆菌接种在7H9添加OADC液体培养基中37℃培养6～8 d,细菌密度为1×108/mL～2×108/mL。抗痨颗粒浸提物的最低抑菌浓度(MIC)为0.638 8 g·L-1。加到液体培养基中,作用20 h,终浓度为相当于生药0.255 6 g·mL-1。局部内环境与外部环境完全隔离下,采用双向凝胶电泳分离全菌总蛋白,得到差异表达的蛋白质点进行分析比对。结果:耐多药结核分枝杆菌用药前后,发现了8个差异斑点,确定其中2个蛋白质为:硫代硫酸硫转移酶(thiosulfate sulfurtransferase)和假定蛋白Rv0634A。结论:揭示了抗痨颗粒对耐多药结核分枝杆菌的翻译,结构组成,氨基酸代谢,氰化物和H2S的解毒以及硫和铁转移等各方面均有一定影响,从而破坏结核分枝杆菌的生活能力,从而达到抑制结核分枝杆菌的目的。Objective： To reveal the mechanism of drug action to compare and analyze of the effect of anti-tuberculosis particles extract on multi-drug resistant-mycobacterium tuberculosis proteomics to reveal the mechanism of drug action.Method：The clinical mycobacterium tuberculosis with multidrug resistance was added in Middlebrook 7H9 OADC liquid medium at 37 ℃ for 6-8 d,when the bacterial density researched 1×108/mL-2×108/mL,the minimum inhibitory concentration（MIC） of anti-tuberculosis particulate extracts was 0.638 8 g·L-1,the final concentration was equivalent to 0.255 6 g·mL-1.Local environment and external environment was completely isolated,two-dimensional gel electrophoresis separation of whole cell total protein was used to analyze differentially expressed protein spots.Result： Before and after treatment by anti-tuberculosis particles extract,multidrug-resistant mycobacterium tuberculosis showed eight differences spots,including two determined proteins： thiosulfate sulfur transferase and hypothetical protein Rv0634A.Conclusion： The results indicate that anti-tuberculosis particles have an impact on the translation,composition,amino acid metabolism,cyanide,H2S detoxification as well as sulfur and iron transfer of the multi-drug resistant mycobacterium tuberculosis,inhibition of mycobacterium tuberculosis may be contributed to undermining the livelihood of the mycobacterium tuberculosis ability.

关 键 词：抗痨颗粒 耐多药结核分枝杆菌 蛋白质组 硫代硫酸硫转移酶 假定蛋白Rv0634A 

分 类 号：R285.5[医药卫生—中药学]