本文選題：ARDS + 膿毒血癥��； 參考：《中南大學(xué)》2014年博士論文

【摘要】：第一部分膿毒癥ARDS患者病程不同時(shí)期肺泡巨噬細(xì)胞蛋白質(zhì)組學(xué)的變化 背景與目的急性呼吸窘迫綜合征(acute respiratory distress syndrome,ARDS)是一種肺內(nèi)過(guò)度性、失控性的炎癥反應(yīng),多種炎癥細(xì)胞和炎癥介質(zhì)是ARDS發(fā)生發(fā)展的關(guān)鍵因素。肺泡巨噬細(xì)胞(alveolar macrophage, AM)是機(jī)體重要的炎性細(xì)胞,通過(guò)研究在ARDS病程不同時(shí)期中AM蛋白質(zhì)表達(dá)的變化,可能為闡明ARDS發(fā)病機(jī)理提供新的線索和思路。 方法以14個(gè)繼發(fā)于膿毒血癥的ARDS患者為研究對(duì)象,分別于并發(fā)中度ARDS后病程的第一天(第一組)及病情明顯好轉(zhuǎn)的第五天(第二組)取支氣管肺泡灌洗液,從中提取肺泡巨噬細(xì)胞,應(yīng)用雙向凝膠電泳(Two dimensional electrophoresis,2DE)和基質(zhì)輔助激光解析電離飛行時(shí)間質(zhì)譜(Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry, MALDI-TOF-MS)技術(shù)篩選鑒定出差異蛋白質(zhì)。 結(jié)果兩組AM共發(fā)現(xiàn)135個(gè)顯著不同的點(diǎn),其中87個(gè)點(diǎn)容積的變化2.5倍,48個(gè)點(diǎn)顯示2.5倍的變化。進(jìn)一步通過(guò)串聯(lián)質(zhì)譜確定差異蛋白質(zhì)的身份。最終有35個(gè)蛋白質(zhì)得以確認(rèn)。與第一組相比較,第二組中表達(dá)上調(diào)的蛋白質(zhì)點(diǎn)有21個(gè),表達(dá)下調(diào)的蛋白質(zhì)點(diǎn)有14個(gè)。 結(jié)論膿毒癥ARDS患者病程不同時(shí)期肺泡巨噬細(xì)胞蛋白質(zhì)存在差異性表達(dá)。 第二部分膿毒癥ARDS患者病程不同時(shí)期肺泡巨噬細(xì)胞源性HSP27及S100A9、TNF-a、IL-1ra表達(dá)的差異 背景本研究的第一部分選擇了繼發(fā)于膿毒血癥的ARDS患者,在病程不同時(shí)期取肺泡巨噬細(xì)胞進(jìn)行蛋白質(zhì)組學(xué)分析,研究肺泡巨噬細(xì)胞的蛋白質(zhì)組學(xué)的動(dòng)態(tài)變化,通過(guò)串聯(lián)質(zhì)譜最終有35個(gè)蛋白質(zhì)得以確認(rèn),其中表達(dá)上調(diào)的蛋白質(zhì)點(diǎn)有21個(gè),表達(dá)下調(diào)的蛋白質(zhì)點(diǎn)有14個(gè)。本部分研究選擇表達(dá)下調(diào)的蛋白質(zhì)之一熱休克蛋白27(heat shock protein27, HSP27)以及表達(dá)上調(diào)的蛋白質(zhì)S100A9、腫瘤壞死因子-a(Tumor Necrosis Factor-a, TNF-a).白細(xì)胞介素-1受體拮抗劑(Interleukin-1receptor antagonist, IL-1ra)行Western Blot實(shí)驗(yàn)對(duì)第一部分結(jié)果進(jìn)行驗(yàn)證。 目的探討膿毒癥ARDS患者在病程的不同時(shí)期HSP27及S100A9、TNF-a, IL-lra在肺泡巨噬細(xì)胞是否有表達(dá)及表達(dá)是否有差異。 方法以5個(gè)膿毒癥ARDS患者為研究對(duì)象,分別于/ARDS發(fā)病的第一天及第五天行支氣管肺泡灌洗,從中提取肺泡巨噬細(xì)胞,采用Western Blot檢測(cè)ARDS病程不同時(shí)期肺泡巨噬細(xì)胞HSP27及S100A9、TNF-a、IL-1ra的差異表達(dá)。 結(jié)果TNF-α、IL-1ra、S100A9及HSP27在ARDS患者的肺泡巨噬細(xì)胞中均有表達(dá),表達(dá)率100%。TNF-α、IL-lra、S100A9在第二組(ARDS病程第5天)比第一組(ARDS病程第1天)表達(dá)明顯增強(qiáng)：TNF-α表達(dá)相對(duì)值分別是dayl (0.41±0.03), day5(0.74±0.07) P0.05; IL-lra:dayl (0.26±0.04), day5(0.54±0.06) P0.05; S100A9表達(dá)相對(duì)值分別是：dayl (0.29±0.02), day5(0.48±0.03) P0.05。而HSP27在第二組(ARDS病程第5天)比第一組(ARDS病程第1天)表達(dá)明顯減弱,表達(dá)相對(duì)值分別是dayl(0.60±0.04), day5(0.35±0.02), P0.05。兩組間TNF-α、IL-1ra、S100A9及HSP27表達(dá)的差異與經(jīng)過(guò)2-DE分析后的蛋白質(zhì)組學(xué)結(jié)果一致。 結(jié)論膿毒癥ARDS患者在病程的不同時(shí)期HSP27及TNF-α、IL-lra、S100A9在肺泡巨噬細(xì)胞均有差異性表達(dá),且與蛋白質(zhì)組學(xué)結(jié)果一致。 第三部分膿毒癥ARDS患者血漿及支氣管肺泡灌洗液中TNF-a、IL-1、IL-Ira、IL-10、vWF及RAGE的水平及對(duì)ARDS病情發(fā)展及預(yù)后的評(píng)估 背景ARDS是以突出的肺部損傷、炎癥及微血管通透性增加為特征進(jìn)而導(dǎo)致彌漫性肺水腫和機(jī)械性功能障礙,最終導(dǎo)致呼吸衰竭的一種疾患。ARDS患者的主要死因是由于肺損傷引起的系統(tǒng)性炎癥所導(dǎo)致的多器官功能衰竭。在本研究的第一部分獲得了在ARDS病程不同時(shí)期肺泡巨噬細(xì)胞差異性表達(dá)的蛋白質(zhì)。在本研究的第二部分,用Western Blot的研究方法證實(shí)ARDS患者在病程的不同時(shí)期TNF-a及IL-1ra在肺泡巨噬細(xì)胞有差異性表達(dá),且與蛋白質(zhì)組學(xué)結(jié)果一致。因此在第一、第二部分研究的基礎(chǔ)上,本部分研究選擇在ARDS患者血漿及肺泡灌洗液中檢測(cè)TNF-a、IL-1ra及其他炎癥介質(zhì)研究其對(duì)ARDS病情發(fā)展及預(yù)后的預(yù)測(cè)價(jià)值。 目的探討ARDS患者血漿及支氣管肺泡灌洗液(BALF)中TNF-a、IL-1、IL-1ra、IL-10、晚期糖基化終末產(chǎn)物受體(receptor for advanced glycation end products,RAGE)、von Villebrand因子(vWF)對(duì)ARDS病情發(fā)展及預(yù)后的預(yù)測(cè)作用。 方法研究對(duì)象選擇膿毒癥ARDS患者54例,所有研究對(duì)象收住ICU時(shí)均處于病情的急性期,并發(fā)ARDS24小時(shí)以?xún)?nèi),在收住ICU病房的第1天6小時(shí)內(nèi)立即收集外周血液標(biāo)本和肺泡灌洗液(BALF)標(biāo)本。用酶聯(lián)免疫吸附試驗(yàn)測(cè)定細(xì)胞因子TNF-a、IL-1、IL-1ra、IL-10、vWF及RAGE的濃度以研究這些參數(shù)對(duì)ARDS發(fā)展及預(yù)后的預(yù)測(cè)作用。研究對(duì)象分為三組：A組(n=20)為中度ARDS患者組；B組(n=17)為輕度ARDS患者組,該組患者在48小時(shí)內(nèi)發(fā)展為中度ARDS;C組(n=17)為輕度ARDS患者組,該組患者未發(fā)展為中度ARDS。 結(jié)果輕度ARDS患者血漿及灌洗液中TNF-a、IL-1、及RAGE、vWF均是病情進(jìn)展惡化很好的陰性預(yù)測(cè)值,其中血漿中以上各指標(biāo)的陰性預(yù)測(cè)值為83%-95%,灌洗液中88%-94%。血漿中IL-1及vWF的水平和灌洗液中IL-1的水平與死亡正相關(guān),隨著IL-1及vWF的水平增高,死亡風(fēng)險(xiǎn)增高。血漿IL-10、IL-1ra水平與死亡呈負(fù)相關(guān),血漿IL-10、IL-lra水平越高,死亡風(fēng)險(xiǎn)越低。 結(jié)論血漿及肺泡灌洗液中的細(xì)胞因子TNF-a、IL-1、IL-1ra、IL-10及RAGE、vWF對(duì)膿毒癥ARDS患者病情進(jìn)展及預(yù)后有一定的預(yù)測(cè)作用。
[Abstract]:Proteomics changes of alveolar macrophages in patients with ARDS after first partial sepsis

BACKGROUND & OBJECTIVE Acute respiratory distress syndrome ( ARDS ) is a key factor in the development of ARDS after acute respiratory distress syndrome ( ARDS ) .

Methods The alveolar macrophages were extracted from 14 ARDS patients who were secondary to sepsis . The alveolar macrophages were extracted from alveolar macrophages by two dimensional electrophoresis ( 2DE ) and matrix assisted laser desorption / ionization time of Flight Mass Spectrometry ( MALDI - TOF - MS ) .

Results There were 135 significant differences between the two groups . The changes of 87 points were 2.5 times and 2.5 times . The identity of the difference protein was determined by tandem mass spectrometry . The final 35 proteins were confirmed . There were 21 protein spots up - regulated in the second group compared with the first group , and 14 of the down - regulated protein spots were expressed .

Conclusion There is different expression of alveolar macrophage protein in patients with sepsis ARDS .

The difference of expression of HSP27 , TNF - a and IL - 1ra in alveolar macrophages during different stages of ARDS in patients with second partial sepsis

Background : The first part of this study selected ARDS patients who were secondary to sepsis , studied the proteomic analysis of alveolar macrophages in different stages of the course of the disease , studied the dynamic changes of protein groups in alveolar macrophages , and confirmed that there were 21 protein spots up - regulated by tandem mass spectrometry . The first part of IL - 1receptor antagonist ( IL - 1ra ) was verified by Western Blot .

Objective To investigate the expression and expression of HSP27 ( HSP27 ) , TNF - a ( TNF - a ) and interleukin - lra ( IL - lra ) in alveolar macrophages in ARDS patients .

Methods The alveolar macrophages were extracted from the first day and the fifth day of the onset of ARDS in five sepsis ARDS patients , and the difference of HSP27 in the alveolar macrophages and the levels of TNF - a and IL - 1ra in the alveolar macrophages in different stages of ARDS were detected by Western Blot .

Results The relative values of TNF - 偽 , IL - 1ra , S100 A9 and HSP27 were significantly increased in the second group ( 0 . 41 鹵 0 . 03 ) , day5 ( 0 . 74 鹵 0 . 07 ) P 0 . 05 ; IL - lra : dayl ( 0.26 鹵 0.04 ) , day5 ( 0.54 鹵 0.06 ) P 0 . 05 ; the relative values of HSP27 expression were dayl ( 0 . 60 鹵 0.04 ) , day5 ( 0 . 35 鹵 0.02 ) , P 0 . 05 .

Conclusion HSP27 and TNF - 偽 , IL - lra and S100 A9 were differentially expressed in alveolar macrophages in ARDS patients at different stages and were consistent with the results of Proteomics .

The levels of TNF - a , IL - 1 , IL - Ira , IL - 10 , VWF and RAGE in plasma and bronchoalveolar lavage fluid in patients with third partial sepsis ARDS and the evaluation of the development and prognosis of ARDS

In the second part of this study , we studied the expression of TNF - a , IL - 1ra and other inflammatory mediators in the plasma and alveolar lavage fluid of ARDS patients , and the predictive value of TNF - a , IL - 1ra and other inflammatory mediators on the development and prognosis of ARDS patients was studied .

Objective To investigate the prognostic significance of TNF - a , IL - 1 , IL - 1ra , IL - 10 , advanced glycosylation end products ( RAGE ) and von Wilk factor ( VWF ) on the development and prognosis of ARDS patients .

Methods 54 cases of ARDS patients with sepsis were studied . All the subjects were in the acute stage of the condition in the ICU , and the peripheral blood samples and BALF samples were collected immediately within 6 hours of the first day of the intensive care unit . The concentrations of cytokines TNF - a , IL - 1 , IL - 1ra , IL - 10 , von Wilvand RAGE were measured by enzyme - linked immunosorbent assay . The results were divided into three groups : group A ( n = 20 ) as the group of moderate ARDS patients ;
Group B ( n = 17 ) was a mild ARDS group and the group developed into moderate ARDS within 48 hours ; Group C ( n = 17 ) was mild ARDS patients and the group did not develop into moderate ARDS .

Results TNF - a , IL - 1 , IL - 1 , IL - 1 , IL - 1 and IL - 1 levels in plasma and lavage fluid of patients with mild ARDS were negative predictive values . The levels of IL - 1 , IL - 1ra and IL - 1 in plasma were negatively correlated with death . The higher the levels of IL - 10 and IL - 1ra in plasma , the higher the level of IL - 10 and IL - lra , the lower the risk of death .

Conclusion TNF - a , IL - 1 , IL - 1ra , IL - 10 and RAGE in plasma and alveolar lavage fluid can predict the progression and prognosis of ARDS patients .
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R459.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Heat shock protein 27 regulates oxidative stress-induced apoptosis in cardiomyocytes:mechanisms via reactive oxygen species generation and Akt activation[J];Chinese Medical Journal;2007年24期

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本文編號(hào)：2072131