發(fā)布時間：2018-03-21 17:02

  本文選題：燒傷　切入點：肺水腫　出處：《蚌埠醫(yī)學院》2017年碩士論文　論文類型：學位論文

【摘要】：研究背景及目的在嚴重燒傷后早期,微血管擴張,血管通透性增加,大量水分及營養(yǎng)物質(zhì)丟失,炎癥介質(zhì)入侵,嚴重可引起休克及多臟器損傷,而肺臟是最常損傷的器官。燒傷后引起急性肺損傷(ALI)可能機制如下:1.中性粒細胞作為ALI發(fā)病過程中最為重要的細胞,通過釋放活性氧、多種細胞因子及蛋白水解酶引起肺損傷;2.氧化與抗氧化反應的嚴重失衡可能是ALI發(fā)生的機制;3.ALI患者凝血因子發(fā)生異常,繼而導致凝血與抗凝機制作用異常,造成肺泡內(nèi)纖維蛋白沉積;4.肺毛細血管內(nèi)皮細胞的損傷是ALI發(fā)病過程中的中心環(huán)節(jié),肺微血管通透性增加是早期肺損傷的重要原因。粒細胞巨噬細胞集落刺激因子(Granulocyte macrophage colony-stimulating factor,GM-CSF)是一種具有多種功能的細胞因子,其與GM-CSF受體(GMR)結(jié)合促進多種細胞的增殖和分化,保護并提高多類細胞的功能,包括內(nèi)皮細胞、成纖維細胞、中性粒細胞/單核細胞、樹突狀細胞及角質(zhì)形成細胞等。在過去,GM-CSF主要應用于放化療及骨髓移植后的白細胞減少癥、再生障礙性貧血等免疫缺陷性疾病的治療,發(fā)揮抗腫瘤及抗感染作用;現(xiàn)在GM-CSF還可以應用到各類創(chuàng)面的修復,通過加速傷口上皮化、參與新生血管的形成及傷口的收縮以促進傷口愈合;近年來,我們的研究還發(fā)現(xiàn)GM-CSF可以調(diào)節(jié)血管周細胞增加Ang-1的表達,降低VEGF的表達,從而調(diào)節(jié)微血管通透性,促進傷口愈合。綜合以上研究基礎,我們設想GM-CSF可能對嚴重燒傷后的急性肺損傷具有保護作用。本課題在嚴重燒傷急性肺損傷模型的基礎上,腹腔注射GM-CSF,觀察GM-CSF對燒傷后急性肺損傷治療效果,并分析其可能的機理。方法SD大鼠30只隨機分為A、B、C三組,A組為假燙組,其余制模成30%Ⅲ度燒傷,B為立即復蘇用藥組,C為立即復蘇不用藥組,(復蘇液為乳酸林格氏液,按照Parkland公式;GMCSF用藥方法60ug/kg,腹腔內(nèi)注射)各組大鼠均于傷后48h后取外周血,用ELIAS方法檢測MPO、TNFa、IL-6,VEGF、ANG-1;取肺組織檢測肺組織濕干比并做病理檢測,用免疫熒光法,檢測肺組織MPO、IL-6、VEGF及ANG-1表達情況。上述結(jié)果均用t檢驗做統(tǒng)計學分析。結(jié)果1.大鼠燒傷48h后,各組肺組織濕干比均較A組(4.3502±0.0782)明顯增加。其中B組濕干比(4.5834±0.1030)低于C組(4.8509±0.1896),比較差異有顯著統(tǒng)計學意義(P0.01)。2.燒傷48h后檢測各組肺組織MPO濃度含量情況:C組(144.6±6.1325pg/ml)明顯高于B(97.7±4.7603)組,差異有統(tǒng)計學意義(P0.01);A組(68.1±5.7227)略低于B組(97.7±4.7603),差異有統(tǒng)計學意義(P0.05)。3.在建立模型48h后取外周血測得TNF-a、IL-6、VEGF濃度含量:C組(237±18.69pg/ml、124.8±10.8pg/ml、255.7±12.48pg/ml)較A組(135.2±13.17pg/ml、74.5±12.5pg/ml、152.3±8.86pg/ml)及B組(170.2±14.96pg/ml、88.6±8.2pg/ml、179.3±11.2pg/ml)顯著升高,C組明顯高于B組(P0.01),差異有統(tǒng)計學意義。而外周血中B組(3080±218.27pg/ml)ANG-1濃度含量較C組(2045±98.69pg/ml)顯著增高,差異有統(tǒng)計學意義(P0.01),而A組與B組無明顯差異,無統(tǒng)計學意義。4.燒傷48h后肺組織中檢測TNF-a及IL-6的濃度含量C組較A組及B組明顯增加,C組(214.8±8.24pg/ml、116.1±6.0pg/ml)明顯高于B組(177.4±3.78pg/ml、82.1±5.27pg/ml),比較差異有統(tǒng)計學意義(P0.01)。C組肺組織中VEGF濃度含量(279.3±8.07pg/ml)明顯高于及B組(177.4±3.78pg/ml),差異有統(tǒng)計學意義(P0.01);B組ang-1濃度含量(3106±80.2pg/ml)明顯高于C組(2177±93.56pg/ml)(P0.01),差異有統(tǒng)計學意義。5.燒傷48h后對肺組織中的血管通透性指標進行熒光分析檢測,觀察到C組組中IL-6、MPO及VEGF表達含量較A組與B組明顯增多,而ang-1熒光表達A組與B組明顯強于C組。6.燒傷48h后C組Evans Blue OD值(0.318±0.337)明顯高于B組(0.202±0.397),差異有統(tǒng)計學意義(P0.01)。結(jié)論1.大鼠嚴重燒傷后早期,肺組織水腫為早期并發(fā)癥,通過腹腔注射GM-CSF用藥發(fā)現(xiàn)可以減輕肺水腫,提示GM-CSF對嚴重燙傷后急性肺損傷有保護作用。2.GM-CSF通過促進血管外周細胞ANG-1的表達,降低VEGF作用于血管內(nèi)皮細胞通透性,增強細胞與細胞之間的緊密性,減少水分及大分子物質(zhì)的滲出,降低肺毛細血管通透性,對肺部具有保護作用。
[Abstract]:Background and objective in after severe burns, microvascular expansion, increased vascular permeability, large amounts of water and nutrient loss, inflammation invasion, serious can cause shock and multiple organ damage, and lung is the most common organ injury. Burns caused by acute lung injury (ALI) mechanism of neutrophils are as follows: 1. as the most important cell in the pathogenesis of ALI, through the release of reactive oxygen species, a variety of cytokines and proteolytic enzymes caused by lung injury; 2. serious imbalance of oxidation and antioxidation reaction may be the mechanism of ALI; 3.ALI patients with abnormal blood coagulation factor, then lead to the effect of coagulation and anticoagulation mechanism caused by abnormal alveolar fibrin deposition; 4. pulmonary capillary endothelial cell injury is the central link in the pathogenesis of ALI, increase the pulmonary microvascular permeability is an important reason for early lung injury induced by granulocyte macrophage colony. Colony stimulating factor (Granulocyte macrophage, colony-stimulating factor, GM-CSF) is a multifunctional cytokine, and GM-CSF receptor (GMR) combined with the proliferation and differentiation of a variety of cells, protect and improve many types of cells, including endothelial cells, fibroblasts, neutrophils / monocytes, dendritic cells and keratinocytes. In the past, GM-CSF is mainly used in chemotherapy and bone marrow transplantation in the treatment of leukopenia, aplastic anemia and other immunodeficiency diseases, anti-tumor and anti infection effect; now GM-CSF can also be used to repair all kinds of wounds, through accelerated wound epithelialization, formation and participate in wound neovascularization contraction to promote wound healing; in recent years, we have found that GM-CSF can regulate the perivascular cells increase the expression of Ang-1 and decreased the expression of VEGF, from The regulation of microvascular permeability, promote wound healing. Based on the above research foundation, we assume that GM-CSF may have a protective effect on acute lung injury after severe burns. Based on the model of severe burn acute lung injury, intraperitoneal injection of GM-CSF, observe the effect of GM-CSF on the effect of the treatment of acute lung injury after burn injury, and to analyze the possible the mechanism. Methods 30 SD rats were randomly divided into A, B, C three groups, A group was the sham group, the mould into 30% third degree burns, B for immediate recovery group, C for immediate recovery without medication group (resuscitation fluid for lactated Ringer's solution, according to the Parkland formula; use of GMCSF method 60ug/kg, intraperitoneal injection) in all rats after injury after 48h from peripheral blood, MPO, ELIAS was detected by TNFa, IL-6, VEGF, ANG-1; the detection of lung wet dry ratio and pathological detection by immunofluorescence, detection of lung tissue MPO, IL-6. VEGF and ANG-1 杈炬儏鍐，

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