本文選題：幽門螺桿菌 + 球形體　； 參考：《第三軍醫(yī)大學(xué)》2006年博士論文

【摘要】： 幽門螺桿菌(Helicobacter pylori, Hp)是人類慢性活動(dòng)性胃炎和消化潰瘍的重要原因,與胃癌和胃淋巴樣組織淋巴瘤的發(fā)生也極度相關(guān)。目前人類對(duì)其生物學(xué)特點(diǎn)、致病機(jī)制及其與宿主的關(guān)系等方面均尚未完全闡明。隨著對(duì)Hp在分子水平上的研究不斷深入,人們發(fā)現(xiàn)Hp較其它細(xì)菌表現(xiàn)出更高的生物多樣性,提示該菌面對(duì)復(fù)雜的定居環(huán)境,具有高度的變異性和獨(dú)特的適應(yīng)性機(jī)制。大量研究發(fā)現(xiàn),Hp在各種不利因素的作用下可發(fā)生形態(tài)變異,由螺旋體轉(zhuǎn)變?yōu)榍蛐误w。Hp作為一種專性微需氧菌,在有氧脅迫下更容易發(fā)生適應(yīng)性變異,轉(zhuǎn)變?yōu)橐环N有活力但不能培養(yǎng)成活(viable but noculturable stage,VNC)的Hp球形變異體,其在Hp感染的傳播途徑及其相關(guān)消化系統(tǒng)疾病的復(fù)發(fā)中可能發(fā)揮著重要作用。研究Hp在有氧脅迫下發(fā)生適應(yīng)性球形變異的變異特征及其變異本質(zhì),解析Hp的適應(yīng)性調(diào)節(jié)元件,將為揭示Hp可能的傳播途徑、闡明Hp的適應(yīng)性變異機(jī)制,以及創(chuàng)新性地開(kāi)展基因工程改造Hp菌株耐氧培養(yǎng)的研究提供有力的實(shí)驗(yàn)依據(jù)。Hp菌26695株和J99株全基因組測(cè)序工作的完成,為后續(xù)的蛋白質(zhì)組學(xué)研究提供了條件。近幾年,國(guó)內(nèi)外研究者采用蛋白質(zhì)組學(xué)技術(shù)相繼在Hp的遺傳變異、生理、免疫靶點(diǎn)篩選及毒力因子確定等方面進(jìn)行了一系列的研究,而在有氧脅迫下Hp發(fā)生適應(yīng)性球形變異方面卻未見(jiàn)相關(guān)研究報(bào)道。 本研究選擇Hp臨床分離株,采用高氧濃度的培養(yǎng)條件,有氧脅迫Hp發(fā)生球形變異,建立穩(wěn)定的球形體模型,進(jìn)而對(duì)Hp球形體的超微結(jié)構(gòu)、菌體代謝、酶活特性、毒力因子mRNA水平表達(dá)差異、體內(nèi)外回復(fù)定植力等變異特征進(jìn)行實(shí)驗(yàn)研究。同時(shí),采用目前蛋白質(zhì)組學(xué)研究中的核心技術(shù)———雙向電泳,對(duì)適應(yīng)性變異前后的Hp螺旋體和球形體進(jìn)行全菌蛋白質(zhì)表達(dá)譜的比較分析,再通過(guò)MALDI-TOF質(zhì)譜技術(shù)對(duì)差異蛋白進(jìn)行初步鑒定分析,最后通過(guò)基因克隆技術(shù)構(gòu)建差異蛋白相關(guān)基因的Hp高表達(dá)菌株,驗(yàn)證該基因在有氧脅迫條件下對(duì)Hp氧耐受性的作用。具體研究結(jié)果如下:
[Abstract]:Helicobacter pylori (HP) is an important cause of chronic active gastritis and peptic ulcer, which is also closely related to the occurrence of gastric cancer and lymphatic tissue lymphoma. At present, the biological characteristics, pathogenetic mechanism and its relationship with the host have not been fully elucidated. With the further study of HP at molecular level, it has been found that HP exhibits higher biodiversity than other bacteria, suggesting that HP has a high degree of variability and unique adaptive mechanism in the face of complex settlement environment. A large number of studies have found that the morphological variation of HP can occur under the action of various unfavorable factors, from spirulina to spheroid.Hp, as a specific microaerobic bacterium, is more susceptible to adaptive variation under aerobic stress. It may play an important role in the transmission of HP infection and the recurrence of digestive diseases associated with HP infection. To study the variation characteristics and nature of adaptive spherical variation of HP under aerobic stress, to analyze the adaptive regulatory elements of HP, to reveal the possible transmission pathway of HP, and to clarify the adaptive variation mechanism of HP. And the innovative research on the oxygen tolerance culture of genetically engineered HP strain provides a powerful experimental basis. The complete genome sequencing of 26695 and J99 strains of HP strain provides the conditions for further proteomics research. In recent years, researchers at home and abroad have carried out a series of studies on the genetic variation, physiology, immune target screening and virulence factor determination of HP by proteomics. However, there were no related studies on the adaptive globular variation of HP under aerobic stress. In this study, the clinical isolates of HP were selected. Under the condition of high oxygen concentration, the spherical variation of HP was observed under aerobic stress, and a stable spheroidy model was established, and the ultrastructure, cell metabolism and enzyme activity of HP spheroidium were further studied. The expression of virulence factor mRNA and the ability of colonization in vivo and in vitro were studied experimentally. At the same time, the protein expression profiles of Helicobacter pylori and spheroids before and after adaptive mutation were compared and analyzed by two-dimensional electrophoresis, which is the core technique in proteomics research. The differential protein was identified and analyzed by MALDI-TOF mass spectrometry. Finally, the high expression strain of the differential protein related gene was constructed by gene cloning technique to verify the effect of the gene on the oxygen tolerance of HP under aerobic stress. The specific findings are as follows:
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2006
【分類號(hào)】：R378

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 王亮;IL-9重組表達(dá)及其單克隆抗體的制備[D];重慶理工大學(xué);2012年

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本文編號(hào)：2040319