【作者】 何颖；

【导师】 谢建平；

【作者基本信息】 西南师范大学 ， 生物化学与分子生物学， 2005， 硕士

【摘要】 结核病仍然是威胁人类健康的主要杀手之一,每年全球有8百万人感染结核病,3百万人死于结核病。结核分枝杆菌(Mycobacterium tuberculosis,MTB)是结核病的致病菌。有效的抗结核病药物异烟肼等的问世曾对结核病的控制、治疗产生极大的推动作用,但近年来,MTB的多耐药株的增加和与HIV的共感染为结核病的治疗和控制造成了巨大的困难,使得寻求有效的防治结核病的药物成为当务之急。 中药的多靶点作用为结核病的治疗开辟了新的方向,寻找其中的有效成分和发现药物作用的靶标能为控制和治疗结核病提供重要的线索。猫爪草Radix Ranuncoli Ternati系毛茛科植物小毛茛Ranuculus Fernatus Thunb.的干燥块根,在中医临床上用于治疗肺结核、颈淋巴结结核、咽喉炎、肿瘤等。因该药成分复杂,目前对其真正控制结核病的作用机制并不太清楚。蛋白质组学的研究方法可以从生物的蛋白全局变化进行分析,有利于对药物作用机理进行更深入的研究,并发现新的药物靶标。 本研究利用二维凝胶电泳技术对猫爪草作用前后的结核分枝杆菌的全细胞蛋白表达谱进行研究。在药物对结核分枝杆菌作用分析中,通过对药物作用前后结核分枝杆菌的二维电泳图谱的比较和分析,发现了产生差异的22个蛋白质斑点,利用基质辅助激光解吸/电离飞行时间质谱技术对其中的8个蛋白(7个下调和1个上调)进行分析鉴定,获得8个明确的肽质量指纹图谱,通过在数据库中进行检索分析,确定这8个蛋白质分别为硫代硫酸硫转移酶,延长因子Ts,热休克蛋白X,S-腺苷甲硫氨酸合成酶,吲哚-3-甘油磷酸合酶,烯酰-CoA水合酶、琥珀酰辅酶A合成酶和60kD的分子伴侣2(其中除60kD的分子伴侣2上调外其余蛋白均下调表达)。 猫爪草作用前后的结核分枝杆菌的全细胞蛋白表达变化揭示了猫爪草对结核分枝杆菌的潜伏感染,转录、翻译,结构组成,氨基酸、脂肪酸代谢,能量代谢,氰化物的解毒,硫和铁转移以及耐酸性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,达到消灭结核分枝杆菌,治疗结核病的目的。同更多还原

【Abstract】 Tuberculosis remains a major global threat to human heath, with 8 million cases of clinical tuberculosis and 3 million deaths annually. Mycobacterium tuberculosis (MTB) is the causative agent of tuberculosis. The effective anti-tuberculosis drug armazide had promoted the control and treatment of tuberculosis. But recently the increasing of multi-drug resistant strains of MTB and co-infection with the human immunodeficiency virus (HIV) make the treatment and control of tuberculosis more difficult. It is urgent to find novel effective prevention and treatment of tuberculosis.The muti-targets effect of the traditional Chinese medicine implicates a new possible way for the treatment of tuberculosis. Radix Ranuncoli Ternati is the dry root tuber of Ranuculus Fernatus Thunb., which was used to treat pulmonary tuberculosis, lymphoid tuberculosis, pharyngolaryngitis, tumour in traditional Chinese medicine clinic therapeutics. Because the components of the drug is very complex, the mechanism of its therapeutics of tuberculosis remains unclear. The proteomics method can reveal the global proteins of organisms, and is helpful for further research in the mechanism of this drug and the new drug targets.Two-dimensional electrophoresis (2-DE) was the mainstay technology of the proteomics, and was employed to compare the global protein changes between Mycobacterium tuberculosis treated and that untreated with Radix Ranuncoli Ternati. 22 protein spots were revealed as significantly changed before and after the TCM treatment. Eight protein spots which remarkably increased or decreased (7 decreased and 1 increased ) in MTB treated with Radix Ranuncoli Ternati were subject to matrix更多还原