本文選題：急性肺損傷 + 脂肪干細(xì)胞　； 參考：《鄭州大學(xué)》2017年碩士論文

【摘要】：研究背景:急性肺損傷(acute lung injury,ALI)/急性呼吸窘迫綜合征(acute respiratory distress syndrome,ARDS)繼發(fā)于嚴(yán)重感染、創(chuàng)傷、誤吸、缺血再灌注、酸中毒等原因,失控的炎癥反應(yīng)二次打擊后肺往往為最早受損的器官,是ICU中常見(jiàn)的危重病之一,進(jìn)一步發(fā)展可出現(xiàn)嚴(yán)重的呼吸衰竭而危及生命。盡管ALl/ARDS病因不同,但有共同的特征是難治性低氧血癥和進(jìn)行性呼吸窘迫,其確切的發(fā)病機(jī)制目前仍未完全闡明。治療也因原發(fā)病不同變得復(fù)雜,目前臨床中主要采用小潮氣量肺保護(hù)性通氣策略、嚴(yán)格的液體控制管理以及CRRT、ECMO等高端儀器的應(yīng)用,療效有限。雖然在過(guò)去的二十年里治療方法及ICU病房的使用對(duì)疾病預(yù)后有所改善,但較高的發(fā)病率和死亡率,治療困難以及高昂的醫(yī)療成本等問(wèn)題一直困擾著醫(yī)療界,成為全球性重要的公共衛(wèi)生問(wèn)題。因此迫切尋求新的治療方法來(lái)改善現(xiàn)狀,近年來(lái)研究發(fā)現(xiàn)具有多向分化潛能和抗炎作用的干細(xì)胞,有望改善這些癥狀。脂肪干細(xì)胞(adipose-derived stem cells,ADSCs)是近年來(lái)新發(fā)現(xiàn)的一種成體干細(xì)胞,具有來(lái)源豐富、取材方便,而且增殖能力強(qiáng)、免疫功能低等優(yōu)點(diǎn)引起人們的廣泛關(guān)注。目前關(guān)于A(yíng)DSCs在脂多糖(lipopolysaccharide,LPS)誘導(dǎo)的ALI方面的研究較少,本研究進(jìn)一步探討ADSCs對(duì)LPS誘導(dǎo)急性肺損傷大鼠炎性因子表達(dá)的影響及其抗炎作用機(jī)制。目的:建立用LPS誘導(dǎo)的大鼠ALI模型,EdU標(biāo)記ADSCs后觀(guān)察在肺部定植情況,觀(guān)察ADSCs對(duì)LPS誘導(dǎo)的ALI大鼠的治療作用及炎性因子的影響,進(jìn)一步探討抗炎作用機(jī)制。方法:雄性SD大鼠70只,按隨機(jī)數(shù)字表法分為對(duì)照組(10只)、LPS組(30只)、LPS+ADSCs組(30只)。ADSCs培養(yǎng)至第4代時(shí)加入EdU培育24h標(biāo)記干細(xì)胞。腹腔注射LPS8mg/kg制備大鼠ALI模型,LPS+ADSCs組于制模30min后尾靜脈注射300μL ADSCs進(jìn)行干預(yù);對(duì)照組無(wú)處理直接取標(biāo)本,其余兩組在6h、24h、72 h取標(biāo)本。股動(dòng)脈取血檢測(cè)動(dòng)脈血氧分壓(PaO2)及乳酸水平;左心室取血,用酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)血清髓過(guò)氧化物酶(MPO)和白細(xì)胞介素-10(IL-10)水平;開(kāi)胸取肺組織,檢測(cè)肺濕/干質(zhì)量(W/D)比值,光鏡下觀(guān)察肺組織病理學(xué)改變,熒光顯微鏡下觀(guān)察肺組織EdU標(biāo)記的干細(xì)胞定植情況,用蛋白質(zhì)免疫印跡試驗(yàn)(Western Blot)檢測(cè)肺組織核轉(zhuǎn)錄因子-κB(NF-κB)表達(dá)。結(jié)果:用EdU標(biāo)記的ADSCs通過(guò)大鼠尾靜脈注射后6h、24h、72h均有不同程度的在A(yíng)LI肺部定植。與對(duì)照組比較,LPS組6h起肺組織損傷較重,肺W/D比值、血乳酸及血清MPO、IL-10水平和肺組織NF-κB表達(dá)均顯著增加,PaO2顯著下降;與LPS組比較,LPS+ADSCs組6h起肺組織損傷程度明顯減輕,肺W/D比值、血乳酸及血清MPO水平和肺組織NF-κB表達(dá)均明顯降低,PaO2顯著增加,72 h已接近正常對(duì)照組水平[肺W/D比值:6 h為5.33±0.29比5.77±0.42,72 h為5.14±0.46比5.43±0.38;血乳酸(mmol/L):6 h為3.6±1.0比5.7±1.1,72 h為3.1±1.0比3.8±1.2;血MPO(μg/L):6 h為1.50±0.90比2.70±1.85,72 h為0.46±0.30比0.71±0.22;血NF-κB(灰度值):6 h為0.40±0.11比0.50±0.09,72 h為0.24±0.03比0.33±0.06;PaO2(mmHg,1 mmHg=0.133 kPa):6 h為78.0±4.1比74.5±3.2,72 h為89.3±9.4比81.9±3.4;均P0.05],IL-10水平僅24 h顯著高于LPS模型組(ng/L:27.75±15.49比17.52±6.56,P0.05)。結(jié)論:ADSCs移植可在A(yíng)LI大鼠肺部定植,能改善氧合升高PaO2及降低血清乳酸水平,減輕血清中炎癥因子及肺部炎癥反應(yīng),可能與抑制NF-κB活性有關(guān)。
[Abstract]:Background: acute lung injury (acute lung injury, ALI) / acute respiratory distress syndrome (acute respiratory distress syndrome, ARDS) is secondary to severe infection, trauma, mistaken aspiration, ischemia reperfusion, acidosis and other reasons, and the lung is often the earliest damaged organ after two hits, which is one of the most common critical diseases in ICU. In one step, severe respiratory failure can occur. Although the causes of ALl/ARDS are different, the common characteristics are refractory hypoxemia and progressive respiratory distress. The exact pathogenesis is still not fully elucidated. The treatment is also complicated because of the different primary diseases. Gas strategy, strict liquid control management, and the application of CRRT, ECMO and other high-end instruments have limited curative effect. Although the treatment methods and the use of ICU in the past twenty years have improved the prognosis of the disease, the higher incidence and mortality, the difficulty of treatment and the high cost of medical treatment have plagued the medical community. As an important public health problem, it is urgent to seek new treatments to improve the status. In recent years, studies have discovered that stem cells with multiple differentiation potential and anti-inflammatory effects are expected to improve these symptoms. Adipose-derived stem cells (ADSCs) is a newly discovered adult stem cell in recent years, with a rich source of origin. The advantages of ADSCs in lipopolysaccharide (LPS) induced ALI are less. The effect of ADSCs on the expression of inflammatory factors in rats with acute lung injury induced by LPS and the mechanism of its anti-inflammatory mechanism are further explored. The rat ALI model induced by LPS was established and the lung colonization was observed by EdU marker ADSCs. The effect of ADSCs on LPS induced ALI rats and the effect of inflammatory factors were observed. The mechanism of anti-inflammatory action was further explored. Methods: 70 male SD rats were divided into group (10), LPS group (30), LPS+ADSCs group (30).ADSCs. 24h labeled stem cells were cultured with EdU in the fourth generation. ALI model of rats was prepared by intraperitoneal injection of LPS8mg/kg, and LPS+ADSCs group was injected with 300 mu L ADSCs for the injection of LPS+ADSCs. The control group had no treatment directly and the other two groups were collected in 6h, 24h, 72 h. The serum levels of myeloperoxidase (MPO) and interleukin -10 (IL-10) were detected by enzyme linked immunosorbent assay (ELISA); lung tissue was detected in the lungs and the ratio of lung wet / dry mass (W/D) was detected. The pathological changes of lung tissue were observed under light microscope. The colonization of EdU labeled stem cells in lung tissue was observed under fluorescence microscope, and protein immuno printing was used. The expression of nuclear factor kappa B (NF- kappa B) in lung tissue was detected by trace test (Western Blot). Results: EdU marked ADSCs was injected in the tail vein of the rat through the rat tail vein, 6h, 24h, 72h all were in the ALI lung colonization. Compared with group LPS, the damage degree of lung tissue in LPS+ADSCs group was significantly reduced, the ratio of lung W/D in LPS+ADSCs group, the ratio of W/D, serum MPO and NF- kappa B in lung tissue were significantly decreased, PaO2 increased significantly, and the 72 h was close to the normal control group [lung W/D ratio: 6 h was 5.33 + 0.29, 5.77 + and 5.14 + 0.46 ratio 5. Blood lactic acid (mmol/L): 6 h is 3.6 + 1, 5.7 + 1.1,72 h is 3.1 + 1 and 3.8 + 1.2, and blood MPO (g/L): 6 h is 1.50 + 0.90 than 2.70 + 1.85,72 h, and NF- kappa B (gray value). It was 89.3 + 9.4 ratio 81.9 + 3.4, P0.05], IL-10 level was only 24 h significantly higher than that of LPS model group (ng/L:27.75 15.49 than 17.52 + 6.56, P0.05). Conclusion: ADSCs transplantation can be used to colonize the lungs of ALI rats. It can improve the increase of oxygenation and decrease the level of serum lactic acid, reduce the inflammatory factors in the serum and the inflammatory reaction in the lungs, and may be related to the inhibition of NF- kappa B activity.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R563.8

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 馮洋;尹文;;阻斷NF-κB信號(hào)途徑防治急性肺損傷的研究進(jìn)展[J];中國(guó)急救醫(yī)學(xué);2014年04期

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本文編號(hào)：1913782