猫爪草提取物对临床分离结核分枝杆菌蛋白质组表达的影响

Effects of the Radix Ranuncoli Ternati extracts on Mycobacterium tuberculosis proteome profiling revealed by 2D electrophoresis

猫爪草已经临床治疗耐药结核病,但其作用机理和有效成分尚不清楚。为研究其可能的作用靶标,采用双向电泳技术比较分析猫爪草提取物作用前后结核分枝杆菌临床分离株的全细胞蛋白表达差异。发现22个蛋白质斑点具有明显差异,对其中3个表达明显下调的蛋白质斑点进行基质辅助激光解吸电离飞行时间质谱分析,获得了肽质量指纹图谱。数据库检索分析确定这3个点代表的蛋白质分别为硫代硫酸硫转移酶,延长因子Ts和热休克蛋白X,分别参与厌氧硫代谢、蛋白质翻译和蛋白质折叠分泌、转录调控等过程。这有助于深入研究猫爪草对结核分枝杆菌的作用机理,也为发现新的抗结核病治疗药物靶标提供了线索。

Radix Ranuncoli Ternati is clinically effective traditional Chinese medicine for multidrug resistant tuberculosis. Its active components and mechanism of action remain unsolved. Two dimensional gel electrophoresis （2- DE）was employed to address this problem. Globlal proteome of Mycobacterium tuberculosis untreated and treated with Radix Ranuncoli Ternati were compared, and 22 protein spots were found to be expressed differentially. 3 protein spots which remarkably decreased in Mycobacterium tuberculosis treated with Radix Ranuncoli Ternati were subjected to matrix assisted laser desorption ionization time of flight mass spectrometry（MALDI-TOF-MS） analysis. The data obtained from peptide mass finger printing were used for database search. The 3 protein spots in gel were identified as cysA2（thiosulfate sulfurtransferase）, tsf（ elongation factor EF-Ts）and hspX （heat shock protein X）. These data provide insights into the changed global protein patterns of Mycobacterium tuberculosis treated with Radix Ranuncoli Ternati and may prove useful for further study in the mechanism in how Radix Ranuncoli Ternati influence the life of Mycobacterium tuberculosis. The differentially expressed proteins may be potential novel antituherculosis drug targets.