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  本文選題：結(jié)核分枝桿菌 + 抗結(jié)核藥物依賴性��； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2007年碩士論文

【摘要】： 結(jié)核菌藥物依賴現(xiàn)象的發(fā)現(xiàn)，給結(jié)核病的防治又提出新的挑戰(zhàn)。結(jié)核菌藥物依賴，則意味著臨床繼續(xù)使用該藥不但治療無效，而且在一定條件下可能促進(jìn)病原菌生長繁殖，使病情加重。因此對結(jié)核菌藥物依賴現(xiàn)象進(jìn)行研究已刻不容緩。 目前藥物依賴現(xiàn)象研究的瓶頸在于這種依賴性在體外不能穩(wěn)定傳代，因此本研究挑選依賴?yán)Ｆ浇Y(jié)核菌進(jìn)行體外連續(xù)傳代實驗，結(jié)果表明其藥物依賴性可穩(wěn)定遺傳約30代。將經(jīng)傳代依賴性完全消失的結(jié)核菌懸液尾靜脈注射感染小鼠，以不同頻率給予利福平灌胃，觀察到部分結(jié)核菌恢復(fù)依賴性。 依賴菌所致肺結(jié)核以病情重、耐多藥、化療效果差和預(yù)后不良為主要臨床特征，遠(yuǎn)比一般耐藥病例危害大。已知結(jié)核分枝桿菌利福平依賴株和耐受株的rpoB基因突變高發(fā)位點基本相同，DNA指紋類型及分布亦無明顯特殊性，表明前者的變異主要為表型變異。因此，依賴現(xiàn)象的獨特表現(xiàn)可能僅源于基因表達(dá)差異。本研究從蛋白水平著手，優(yōu)化了結(jié)核分枝桿菌的蛋白質(zhì)組分析中的二維凝膠電泳技術(shù)，并用于分析利福平依賴株和耐受株總蛋白質(zhì)的差異，獲得了利福平依賴株和耐受株總蛋白質(zhì)表達(dá)譜，鑒定8個顯著差異表達(dá)的蛋白質(zhì)點，獲得7種蛋白質(zhì)：3-氧�；鵞酰基載體蛋白]合酶(3-oxoacyl-ACP synthase)、抗原85B A鏈(chain A，Ag85B)、烯醇基輔酶A水解酶(ECH)、跨膜絲/蘇氨酸蛋白激酶D(STPK D)、假想轉(zhuǎn)移酶(putative transferase)、3-酮酸-輔酶A轉(zhuǎn)移酶β亞單位(βsubunit，，SCOB)、熱休克蛋白HSPX，初步討論了藥物依賴現(xiàn)象的分子機(jī)制。 運用比較蛋白質(zhì)組學(xué)方法有助于研究藥物依賴結(jié)核菌的蛋白質(zhì)表達(dá)譜及差異表達(dá)的蛋白質(zhì)分子，為進(jìn)一步探尋結(jié)核菌藥物依賴性的發(fā)生發(fā)展奠定基礎(chǔ)。
[Abstract]:The discovery of drug dependence of tuberculous bacteria poses a new challenge to the prevention and treatment of tuberculosis. The drug dependence of tuberculous bacteria means that clinical use of the drug is not only ineffective, but also may promote the growth and reproduction of pathogenic bacteria under certain conditions, and make the disease worse. Therefore, it is urgent to study the drug dependence of tuberculous bacteria. At present, the bottleneck of drug dependence research is that the drug dependence can not be subcultured stably in vitro. Therefore, this study selected rifampicin-dependent tuberculous bacilli for continuous passage in vitro. The results showed that drug dependence could be inherited steadily for about 30 generations. Mice infected with Mycobacterium tuberculosis suspension completely disappeared after passage dependence were injected with rifampicin at different frequencies and some of them were observed to recover their dependence. The main clinical characteristics of pulmonary tuberculosis caused by dependent bacteria are serious condition, multidrug resistance, poor chemotherapeutic effect and poor prognosis, which are far more harmful than those of common drug resistant cases. It is known that the high frequency of rpoB gene mutation in rifampicin dependent strain and resistant strain of Mycobacterium tuberculosis has no obvious particularity, indicating that the variation of the former is mainly phenotypic variation. Therefore, the unique manifestation of dependence may be due only to differences in gene expression. In this study, we optimized two-dimensional gel electrophoresis for proteome analysis of Mycobacterium tuberculosis at protein level, and used to analyze the differences of total proteins between rifampicin dependent and tolerant strains. The total protein expression profiles of rifampicin dependent and tolerant strains were obtained and eight differentially expressed protein spots were identified. Seven kinds of protein synthase, 3-oxoacyl-ACP synthase, antigen85B A chain, enol coenzyme A hydrolase, transmembrane silk / threonine protein kinase D(STPK, hypothetical transferase putative transfer enzyme 3-ketoacyl-ACP synthase, antigen 85B A chain, enol coenzyme A hydrolase, transmembrane silk / threonine protein kinase D(STPK, were obtained. The molecular mechanism of drug dependence was preliminarily discussed by 尾 subunit SCOBX, heat shock protein HSPX. The application of comparative proteomics is helpful to study the protein expression profile and differentially expressed protein molecules of drug-dependent tuberculous bacteria and lay a foundation for further exploration of the occurrence and development of drug dependence of tuberculous bacteria.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R378

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