【作者】 高孟秋；

【导师】 朱莉贞；

【作者基本信息】 北京市结核病胸部肿瘤研究所 ， 内科学， 2006， 博士

【摘要】 研究背景 结核病作为一种传染性疾病困扰了人类几千年，虽然近代在规范的联合化疗原则指导下，尤其是利福平问世后，初治肺结核可以达到95％的治愈率，但是与其他感染性疾病相比较，结核病的治疗过程中还存在着病程迁延、易复发的问题，随着人类社会步入老龄化、HIV感染的流行，这一问题日显突出，成为人类控制结核病难以逾越的障碍。因此探索结核病的复发机制，寻求新的有效的抗结核药物成为战胜结核病新的希望。 结核分枝杆菌的持留状态或称持留菌（Persistent Mycobacterium tuberculosis or persister）被认为是导致结核病程迁延、复发的根本原因。低氧状态及营养匮乏条件下，结核分枝杆菌通过启动乙醛酸支路代谢而获得能量，导致异柠檬酸裂解酶（ICL）、16kDa小分子热休克蛋白Acr（Alpha-crystallin chaperone）的高度表达，形态上表现为细胞壁增厚等改变。改变了形态和代谢途径的结核分枝杆菌，逃避了现有抗结核药物对其杀灭作用和机体免疫系统的汰除作用，进一步研究、认识结核分枝杆菌的持留状态的生物学特性对于人类彻底战胜结核病具有重要意义。 研究目的 建立体内外结核分枝杆菌持留菌的模型和临床标本分析，通过培养和检测mRNA表达，以期观察在不同化疗阶段中结核分枝杆菌的状态，评价化疗作用及探讨复发机制。 研究方法 （1）建立体外培养方法并应用定量RT-PCR测定结核分枝杆菌H₃₇R_V在低氧更多还原

【Abstract】 Background:Tuberculosis is an important infectious disease. Although chemotherapy can cure 95% of the new tuberculosis patient, but the long term of the disease, and the relapse are the main barrier of the control of the disease. The persist tuberculosis are believed as the causes. At the microaerophilic or anaerobic conditions, the metabolism of mycobacterium tuberculosis shiftdown to glyoxylate bypass circumvents, and the express of isocitrate lyase and the 16-kilodalton α -crystallin chaperone protein hemologue (Acr) are expressed highly and cell wall thickening in shape. The persister are usually resistant to chemotherapy and it can be able to avoid elimination from the human host by immune system. Objective:At first, to set up a persistence model in vitro and in vivo and observe the metabolic state of TB in different course of chemotherapy by detecting of mRNA. Methods:(1) Compared the levels of mRNA of ICL、 Acr and 85B in different conditions in vitro, and the levels of mRNA were determined by quantitative RT-PCR.(2) Cultured tuberculosis detected the expression of mRNA of the ICL、 Acr and 85 B in Cornell model in different period.(3) Detected the expression of mRNA of ICL、 Acr and 85B in operation materials from 12 patients who had received chemotherapy at least for 3 month.Results:Both ICL and Acr are expressed highly in vitro and vivo models, the ratio of ICL/85Bare concerned with the metabolist state of MTB and we also detected the highly更多还原