【摘要】：背景肺纖維化(pulmonary fibrosis,PF)是目前在呼吸系統(tǒng)中死亡率非常高的一種疾病,誘導(dǎo)其發(fā)病的因素也很多,PF的主要臨床特征有呼吸困難逐漸遞增和肺間質(zhì)纖維增生呈彌漫性。能夠引起PF的發(fā)病因素有很多,由于缺乏有效的藥物來治療,所以其預(yù)后的效果并不十分理想。百草枯(paraquat,PQ)是目前應(yīng)用比較廣泛的一種除草劑,百草枯中毒之后最主要靶點(diǎn)之一就是肺部,人體接觸之后極易在肺部聚積,之后引起的肺水腫、低氧以及肺部急性損傷都可能導(dǎo)致肺纖維化的發(fā)生,但其作用機(jī)制目前還不十分清楚。本研究以百草枯誘導(dǎo)的大鼠肺纖維化模型為研究對(duì)象,分析鑒定大鼠肺纖維化肺組織之間的差異表達(dá)蛋白,以期尋找與肺纖維化相關(guān)的關(guān)鍵蛋白,為探索闡明肺纖維化形成、發(fā)生發(fā)展機(jī)制奠定重要的理論和實(shí)驗(yàn)基礎(chǔ)。目的用同位素標(biāo)記相對(duì)與絕對(duì)定量(iTRAQ)技術(shù)篩選百草枯誘導(dǎo)的大鼠肺纖維化模型特異性表達(dá)蛋白,以期找到一些與肺纖維化相關(guān)的蛋白。方法將20只雄性SD大鼠(180-200g)分為兩組:模型組:灌胃給予50mg/kg百草枯,對(duì)照組:灌胃給予50mg/kg生理鹽水。在第28天時(shí)分別處死兩組大鼠,通過HE染色的方法觀察大鼠肺組織學(xué)的改變;應(yīng)用iTRAQ標(biāo)記聯(lián)合LC-MS/MS質(zhì)譜檢測(cè)分析蛋白的表達(dá)量;利用NCBI數(shù)據(jù)庫和Proteome Discoverer軟件分析蛋白表達(dá)間的差異。結(jié)果HE染色表明實(shí)驗(yàn)組大鼠肺纖維化分級(jí)大于對(duì)照組;iTRAQ技術(shù)分析鑒定出268個(gè)差異表達(dá)蛋白,其中表達(dá)上調(diào)蛋白194個(gè),表達(dá)下調(diào)蛋白74個(gè)。結(jié)論通過篩選百草枯誘導(dǎo)的大鼠肺纖維化模型特異性表達(dá)蛋白,表明iTRAQ標(biāo)記聯(lián)合LC-MS/MS質(zhì)譜技術(shù)可以對(duì)表達(dá)具有差異的蛋白進(jìn)行相關(guān)研究,并通過篩選的方法獲得與肺纖維化的發(fā)生和發(fā)展有關(guān)聯(lián)的蛋白質(zhì),這將對(duì)了解肺纖維化的發(fā)生發(fā)展過程產(chǎn)生十分重大的意義,并且還可能為肺纖維化的治療或者預(yù)后提供理論基礎(chǔ)。
[Abstract]:Background Pulmonary fibrosis (pulmonary fibrosis,PF) is a disease with high mortality in the respiratory system at present. The main clinical characteristics of PF are the gradual increase of dyspnea and the diffuse proliferation of pulmonary interstitial fiber. There are many factors that can cause PF, and the prognosis is not very good because of the lack of effective drugs to treat it. Paraquat (paraquat,PQ) is one of the most widely used herbicides. One of the main targets after paraquat poisoning is lung. Hypoxia and acute lung injury may lead to pulmonary fibrosis, but its mechanism is not clear. In this study, the rat pulmonary fibrosis model induced by paraquat was used to analyze and identify the differentially expressed proteins in the lung tissues of rats with pulmonary fibrosis, in order to find the key proteins related to pulmonary fibrosis and to elucidate the formation of pulmonary fibrosis. The mechanism of occurrence and development lays an important theoretical and experimental foundation. Objective to screen the specific expression proteins of paraquat induced pulmonary fibrosis model by using isotope labeling and absolute quantitative (iTRAQ) technique in order to find some proteins related to pulmonary fibrosis. Methods Twenty male SD rats (180-200g) were divided into two groups: the model group was treated with 50mg/kg paraquat and the control group with 50mg/kg saline. On the 28th day, the rats of the two groups were killed, the changes of lung histology were observed by HE staining, and the expression of protein was detected by iTRAQ labeling and LC-MS/MS mass spectrometry. The differences of protein expression were analyzed by NCBI database and Proteome Discoverer software. Results HE staining showed that the grade of pulmonary fibrosis in the experimental group was higher than that in the control group, and 268 differentially expressed proteins were identified by iTRAQ technique, including 194 up-regulated proteins and 74 down-regulated proteins. Conclusion by screening the specific expression protein of paraquat induced pulmonary fibrosis in rats, it is suggested that iTRAQ labeling combined with LC-MS/MS mass spectrometry can be used to study the expression of differentially expressed proteins. The protein associated with the occurrence and development of pulmonary fibrosis is obtained by screening method, which will be of great significance in understanding the process of occurrence and development of pulmonary fibrosis. It may also provide a theoretical basis for the treatment or prognosis of pulmonary fibrosis.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R-332;R563

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