本文選題：急性肺損傷 + 內(nèi)皮祖細(xì)胞��； 參考：《第三軍醫(yī)大學(xué)》2017年碩士論文

【摘要】：背景與目的:急性肺損傷(acute lung injury,ALI)是多種繁雜病因造成的毛細(xì)血管內(nèi)皮細(xì)胞和肺泡上皮細(xì)胞的損傷,伴有不斷加重的低氧血癥和ARDS。ALI具有病死率高、治療困難、預(yù)后差等特點(diǎn),尋找有效的ALI治療靶點(diǎn)仍然是目前危重病醫(yī)學(xué)亟待解決的關(guān)鍵問(wèn)題之一[1]。H_2S被視為和CO、NO一類的第三種氣態(tài)遞質(zhì)[2]。前期研究發(fā)現(xiàn),在肢體損傷導(dǎo)致的遠(yuǎn)隔肺損傷中,肺H_2S產(chǎn)生減少、循環(huán)H_2S濃度降低,給以外源性H_2S供體Na HS可減輕肢體損傷導(dǎo)致的肺損傷[3]。大量文獻(xiàn)報(bào)道,外源性H_2S可減輕高壓氧、LPS、缺血再灌注、機(jī)械通氣引起的肺損傷[4]。但是,H_2S在LPS誘發(fā)小鼠ALI中發(fā)揮保護(hù)效用的機(jī)制尚未闡明。本研究通過(guò)經(jīng)氣管內(nèi)注射LPS來(lái)構(gòu)建小鼠ALI模型,檢測(cè)炎癥因子、H_2S、內(nèi)皮祖細(xì)胞(endothelialprogenitorcel,EPCs)、肺巨噬細(xì)胞(alveolar macrophage,AM)等在ALI小鼠體內(nèi)的含量變化,觀察H_2S對(duì)于ALI損傷程度的影響,探討:(1)H_2S對(duì)EPCs動(dòng)員和再內(nèi)皮化的作用,(2)H_2S與ALI肺內(nèi)AM表型轉(zhuǎn)化的關(guān)聯(lián),為ALI的治療提供新的解決思路。方法:1.氣管內(nèi)注射LPS構(gòu)建小鼠ALI模型。2.肉眼下比較各組小鼠肺大體標(biāo)本,光學(xué)顯微鏡下比較肺病理學(xué)改變,測(cè)定肺系數(shù)和肺病理學(xué)評(píng)分。3.測(cè)定不同時(shí)相點(diǎn)各組小鼠血清炎癥因子IL-6、IL-8和TNF-α濃度。4.測(cè)定各組小鼠血清H_2S在不同時(shí)相點(diǎn)的濃度。5.檢測(cè)各組小鼠肺組織CD34和VEGFR2的表達(dá),計(jì)數(shù)CD34+/VEGFR2+細(xì)胞數(shù)目。6.檢測(cè)各組小鼠肺組織i NOS和Arginase的表達(dá)。結(jié)果:1.成功建立ALI模型:LPS組各時(shí)相點(diǎn)肺組織呈現(xiàn)不同程度的毛細(xì)血管上皮和肺泡壁破壞、肺泡壁和肺間質(zhì)顯著增厚,肺間質(zhì)及肺泡腔內(nèi)大量紅細(xì)胞滲出和炎性細(xì)胞浸潤(rùn);與Sham組相比,肺系數(shù)、IQA評(píng)分顯著升高。2.LPS組第3天肺大體標(biāo)本的出血面積顯著大于其它時(shí)相點(diǎn),肺組織肺間質(zhì)增厚、紅細(xì)胞滲出和炎性細(xì)胞浸潤(rùn)程度最為嚴(yán)重,肺系數(shù)和IQA病理學(xué)評(píng)分高于其他時(shí)相點(diǎn)。3.Na HS組小鼠肺組織IQA值及肺系數(shù)比LPS組顯著降低,PAG組和AOAA組小鼠肺組織IQA值及肺系數(shù)比LPS組顯著升高。4.ALI小鼠血清H_2S濃度顯著減少,Na HS組血清H_2S濃度比LPS組顯著增加。5.Na HS組小鼠肺組織CD34和VEGFR2表達(dá)較LPS組顯著增多,CD34+/VEGFR2+細(xì)胞數(shù)目顯著增加。6.Na HS組肺組織中Arginase/i NOS比值較LPS組顯著升高。結(jié)論:1.通過(guò)氣管內(nèi)滴注LPS構(gòu)建小鼠ALI模型的方法具有重復(fù)性好,穩(wěn)定性強(qiáng)的優(yōu)點(diǎn)。為進(jìn)一步研究ALI相關(guān)生理病理機(jī)制提供穩(wěn)定的動(dòng)物模型基礎(chǔ)。2.LPS氣管內(nèi)滴注后第3天是ALI小鼠肺損傷最嚴(yán)重的時(shí)間節(jié)點(diǎn),因此作為后續(xù)研究H_2S對(duì)ALI的作用及機(jī)制的時(shí)相點(diǎn)。3.外源性H_2S可通過(guò)促進(jìn)EPCs動(dòng)員及在肺部的再內(nèi)皮化、糾正ALI時(shí)AM的極化失衡,抑制炎癥反應(yīng)、促進(jìn)損傷肺組織愈合,進(jìn)而減輕ALI肺損傷。
[Abstract]:Background & objective: acute lung injury (Ali) is a kind of injury of capillary endothelial cells and alveolar epithelial cells caused by many complicated causes. It is accompanied by increasing hypoxemia and ARDS.ALI with the characteristics of high mortality, difficult treatment and poor prognosis. Finding an effective therapeutic target for ALI is still one of the key problems in critical medicine [1] .Hs / s _ 2S is regarded as the third gaseous transmitter such as CONO _ 2 [2]. Previous studies showed that in the distant lung injury caused by limb injury, H2S production and circulating H2S concentration decreased in the lung. The exogenous H2S donor NaHS could reduce the lung injury caused by limb injury [3]. It is reported that exogenous H2S can reduce lung injury induced by hyperbaric oxygen, ischemia reperfusion and mechanical ventilation. However, the mechanism of the protective effect of H2S in LPS induced ALI in mice has not been elucidated. In this study, the ALI model of mice was established by intratracheal injection of LPS, and the levels of inflammatory factor, endothelial-progenitor cells, endothelial progenitor cells, alveolar macrophage AM) were detected in ALI mice, and the effects of H2S on the degree of ALI damage were observed. To explore the role of H2S in mobilization and re-endothelialization of EPCs and its relationship with the phenotypic transformation of AM in the lung of ALI, and to provide a new solution for the treatment of ALI. Method 1: 1. Establishment of mouse ALI model by intratracheal injection of LPS. Gross lung samples of each group were compared under naked eye, lung pathological changes were compared under optical microscope, lung coefficient and lung pathological score were measured. The levels of IL-6, IL-8 and TNF- 偽 in serum of mice at different time points were determined. The concentration of H 2S in serum of each group of mice was determined at different time points. The expression of CD34 and VEGFR2 in lung tissue of each group was detected, and the number of CD34 / VEGFR2 cells. The expression of I NOS and Arginase in lung tissue of each group was detected. The result is 1: 1. The ALI model was successfully established at different time points in the ALI group. The pulmonary tissue showed various degrees of destruction of capillary epithelium and alveolar wall, significant thickening of alveolar wall and interstitial tissue, massive exudation of erythrocytes and infiltration of inflammatory cells in the interstitial and alveolar lumen of the lung, as compared with that in the Sham group. IQA score of lung coefficient was significantly increased in LPS group. 2. The hemorrhage area of lung gross specimens in LPS group was significantly larger than that in other time points on the 3rd day. The pulmonary interstitial thickening, erythrocyte exudation and inflammatory cell infiltration were the most serious in LPS group. Lung coefficient and IQA pathological score were higher than those of other time point. 3.The lung tissue IQA and lung coefficient of NaHS group were significantly lower than that of LPS group. 4. Serum H2S concentration of Ali mice was significantly higher than that of LPS group. Compared with LPS group, the expression of CD34 and VEGFR2 in lung tissue of Na-HS group was significantly higher than that of LPS group. The number of CD34 / VEGFR2 cells was significantly increased. 6. The ratio of Arginase/i NOS in lung tissue of NaHS group was significantly higher than that of LPS group. Conclusion 1. The method of establishing mouse ALI model by intratracheal instillation of LPS has the advantages of good repeatability and strong stability. In order to provide a stable animal model basis for further study on the physiological and pathological mechanism of ALI. 2.The third day after intratracheal instillation of ALI was the most severe time node of lung injury in ALI mice. Exogenous H2S can correct the imbalance of AM polarization in ALI by promoting EPCs mobilization and pulmonary re-endothelialization, inhibit inflammatory reaction, promote the healing of injured lung tissue, and then alleviate the lung injury of ALI.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R563.8

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本文編號(hào)：1849206