發(fā)布時(shí)間：2018-03-09 20:52

  本文選題：封閉負(fù)壓引流技術(shù)　切入點(diǎn)：燒傷　出處：《第四軍醫(yī)大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：熱能可引起燒傷部位即刻而不可逆性的壞死，鄰近該部位未直接燒傷部位也可產(chǎn)生損傷，而且損傷程度進(jìn)行性加重，不斷加深擴(kuò)大，最終導(dǎo)致組織壞死。這種現(xiàn)象的發(fā)生不僅加重了患者的病情，而且增加燒傷的致殘率及致死率，是燒傷治療面臨的問題之一。封閉負(fù)壓引流(vacuum-assisted closure，VAC)療法是利用負(fù)壓吸引裝置與特殊敷料結(jié)合后，在創(chuàng)面處產(chǎn)生低于大氣壓的壓力，促進(jìn)創(chuàng)面愈合的技術(shù)，已廣泛應(yīng)用于各類創(chuàng)面的治療。有研究證明封閉負(fù)壓引流治療可逆轉(zhuǎn)燒傷瘀滯區(qū)，，減少燒傷創(chuàng)面損傷深度及壞死面積，但缺乏進(jìn)一步的機(jī)制研究。在燒傷瘀滯區(qū)進(jìn)行性壞死過程中，血管內(nèi)皮細(xì)胞既是損傷的主要靶細(xì)胞，也是病理進(jìn)展的效應(yīng)細(xì)胞。有效的保護(hù)血管內(nèi)皮細(xì)胞，減輕其損傷及功能失常是逆轉(zhuǎn)燒傷瘀滯區(qū)的關(guān)鍵。自噬是指細(xì)胞消化自身多余、受損、衰老的蛋白質(zhì)及細(xì)胞器，以維持內(nèi)環(huán)境穩(wěn)定的細(xì)胞機(jī)制。本課題以大鼠梳狀燒傷模型應(yīng)用封閉負(fù)壓引流技術(shù)進(jìn)行治療，從內(nèi)皮細(xì)胞功能和自噬水平的變化，初步探索VAC技術(shù)治療燒傷瘀滯區(qū)的效果和作用機(jī)制，為臨床治療提供理論依據(jù)。實(shí)驗(yàn)共分以下四個(gè)部分： 1.建立大鼠銅梳燒傷模型。銅梳梳齒間未直接燒傷的間隙區(qū)即為瘀滯區(qū)。實(shí)驗(yàn)動(dòng)物隨機(jī)分為VAC組和對(duì)照組。對(duì)瘀滯區(qū)攝像，用圖像分析軟件，測(cè)量燒傷瘀滯區(qū)壞死面積的變化。HE染色觀察瘀滯區(qū)壞死深度。結(jié)果顯示：燒傷6小時(shí)后VAC組燒傷瘀滯區(qū)壞死面積及深度明顯小于對(duì)照組。 2. NO和ET-1含量檢測(cè)。我們用硝酸還原酶法檢測(cè)瘀滯區(qū)組織的一氧化氮（NO）的含量，用ELISA法檢測(cè)內(nèi)皮素-1(ET-1)水平，對(duì)比VAC組和對(duì)照組之間兩者比值的差異，分析VAC對(duì)血管內(nèi)皮細(xì)胞分泌功能的影響。結(jié)果顯示：燒傷后VAC組瘀滯區(qū)組織NO含量明顯多于傷前對(duì)照組，傷后12小時(shí)達(dá)到峰值，72小時(shí)仍高于正常值。對(duì)照組中NO含量低于VAC組，傷后6小時(shí)達(dá)到峰值，12小時(shí)即降低到正常含量以下，并持續(xù)下降。兩組傷后6小時(shí)及其后各時(shí)間點(diǎn)差異有統(tǒng)計(jì)學(xué)差異；燒傷后早期VAC組燒傷區(qū)瘀滯組織中ET-1含量較傷前對(duì)照組明顯上升，傷后24小時(shí)達(dá)到峰值，48小時(shí)后低于正常值。對(duì)照組燒傷瘀滯區(qū)組織中ET-1含量也上升，但上升程度低于VAC組，傷后6小時(shí)達(dá)到峰值，傷后12小時(shí)即降低到正常值以下。兩組傷后6、12、24小時(shí)差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。傷后早期VAC組ET-1/NO比值較低，12小時(shí)到達(dá)最低值后逐步回升，接近正常值。而對(duì)照組ET-1/NO比值持續(xù)升高，在72小時(shí)仍很高。兩組傷后6小時(shí)及其后各時(shí)間點(diǎn)差異均有明顯差異（P＜0.05）。 3. TM（血栓調(diào)節(jié)蛋白）和TM mRNA檢測(cè)。用免疫組化染色和實(shí)時(shí)熒光PCR檢測(cè)內(nèi)皮細(xì)胞TM和TM mRNA表達(dá)，分析VAC治療對(duì)血管內(nèi)皮細(xì)胞抗凝血、抗炎功能的影響。結(jié)果顯示：傷后各時(shí)間點(diǎn)VAC組瘀滯區(qū)組織均可見TM陽性表達(dá)。對(duì)照組傷后12小時(shí)可見TM陽性表達(dá)，但傷后24小時(shí)及其后時(shí)間點(diǎn)均為陰性。傷后各時(shí)間點(diǎn)，兩組TM mRNA表達(dá)均呈下降趨勢(shì)，但VAC組表達(dá)量高于對(duì)照組。傷后6小時(shí)及其后各時(shí)間點(diǎn)差異明顯（P＜0.05）。 4. LC3和Beclin-1表達(dá)檢測(cè)。免疫組化染色觀察燒傷瘀滯區(qū)LC3及Beclin-1的表達(dá)，對(duì)比VAC組與對(duì)照組之間的差異，分析VAC治療對(duì)燒傷瘀滯區(qū)細(xì)胞自噬的影響。結(jié)果顯示: VAC組瘀滯區(qū)LC3和Beclin-1表達(dá)率傷后6至24小時(shí)低于對(duì)照組，傷后48至72小時(shí)高于對(duì)照組，半定量分析顯示兩組在傷后6小時(shí)及其后各時(shí)間點(diǎn)差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。
[Abstract]:Heat can cause burns immediate and irreversible necrosis, adjacent to the site without direct burn can also cause damage, and the damage degree of progressive deepening, expanding, resulting in tissue necrosis. This phenomenon not only aggravate the patient's condition, but also increase the burn rate of morbidity and mortality, is one of the burn treatment problems. VSD (vacuum-assisted closure, VAC) therapy is the use of negative pressure suction device with special dressings combined, produce subatmospheric pressure on the wound, promote wound healing technique has been widely used in the treatment of various wounds. Studies have shown that vacuum sealing drainage treatment can reverse burn stasis stagnation zone, reduce the damage of deep burn wound and necrosis area, but the lack of further studies. In the process of burn necrosis of Stasis Zone, both vascular endothelial cells Is the main target of cell damage, is the pathological progress of effector cells. Vascular endothelial cell protection effectively, reduce the injury and dysfunction is the key to reverse the burn Stasis Zone. Autophagy is refers to the cell to digest their excess, damaged proteins and organelles of aging, to maintain a stable environment within the cellular mechanism of this subject. By using the rat comb burn model of vacuum sealing drainage for treatment, from the changes of endothelial cell function and autophagy, explore the effect and mechanism of VAC technique in the treatment of burn Stasis Zone, provide a theoretical basis for clinical treatment. The experiment is divided into the following four parts:
1. to establish a rat model of burn. Copper comb gap between copper comb comb is not directly burn the Stasis Zone. The experimental animal were randomly divided into VAC group and control group. The camera of the Stasis Zone, using image analysis software to observe the Stasis Zone of necrosis depth changes of.HE measuring burn Stasis Zone of necrosis area staining. The results showed that 6 hours after burn VAC group burn the Stasis Zone of necrosis area and depth significantly less than the control group.
2. the detection of NO and ET-1 content of nitric oxide. We detected by nitrate reductase method the Stasis Zone tissue (NO) were detected by ELISA et -1 (ET-1) level, the difference of the ratio between the two comparison between VAC group and control group, to analyze the effect of VAC on the secretion of vascular endothelial cells. The results showed that burn the content of NO in group VAC was significantly more than the Stasis Zone tissue injury control group before and 12 hours after injury and reached the peak, 72 hours is still higher than normal. NO content of control group was lower than that of VAC group, 6 hours after injury and reached the peak, 12 hours reduced to normal levels, and continued to decline. There were significant differences in 6 hours the two groups after injury and after each time point of difference; early lag in VAC group after burn in burn area and the content of ET-1 is the pre injury control group increased significantly, and reached the peak at 24 hours after injury, 48 hours less than normal. The control group also ET-1 content in blood stasis stagnation area burn The rise, but the rise is less than the VAC group, and reached the peak at 6 hours after injury, 12 hours after injury is reduced to below normal. Two groups of 6,12,24 hours after injury was statistically significant (P < 0.05). Early after injury VAC group ET-1/NO ratio is low, 12 hours to reach the lowest value gradually rise, close to normal value. While the control group, the ratio of ET-1/NO increased in 72 hours is still very high. 6 hours after injury and two groups had significant difference at each time point after the difference (P < 0.05).
3. TM (thrombomodulin). The expression of mRNA and TM detected by immunohistochemical staining and real-time fluorescence PCR detection of endothelial cells TM and TM mRNA analysis of VAC treatment on vascular endothelial cell function of anticoagulant, anti-inflammatory effects. The results showed that the positive expression of TM in the VAC group the Stasis Zone tissue could be seen after injury 12 hours after injury. The control group showed positive expression of TM, but 24 hours after injury and later time points were negative. At each time point, the expression of mRNA TM in two groups were decreased, but the expression of VAC group was higher than the control group. 6 hours after injury and after each time point difference (P < 0.05).
4. LC3 and Beclin-1 expression was detected by immunohistochemical staining. The expression of LC3 and Beclin-1 to observe the burn of the Stasis Zone, the difference between the VAC group and the control group comparison, analysis of effect of VAC treatment on burn cell Stasis Zone of autophagy. The results showed that VAC LC3 and Beclin-1 group the Stasis Zone expression rate after 6 to 24 hours compared with the control group, 48 hours after injury to 72 higher than the control group, semi quantitative analysis showed that the two groups in 6 hours after injury and after each time point had significant difference (P < 0.05).

【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R644

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 許龍順,陳紹宗,喬騁,李學(xué)擁;負(fù)壓對(duì)創(chuàng)面血流量的影響[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2000年08期

2 李金清,陳紹宗,付小兵,李學(xué)擁,許龍順,李望舟;封閉負(fù)壓引流對(duì)豬皮膚軟組織爆炸傷感染創(chuàng)面肉芽組織生成的影響[J];解放軍醫(yī)學(xué)雜志;2004年08期

3 湯蘇陽,陳紹宗,胡昭華,宋梅,曹大勇,呂曉星;封閉式負(fù)壓引流技術(shù)對(duì)大鼠創(chuàng)面愈合過程中VEGF和BCL-2的影響[J];中國(guó)美容醫(yī)學(xué);2003年02期

4 盧興國(guó),付國(guó)勝,黃連生,張佩利;原發(fā)性高血壓血漿血栓調(diào)節(jié)蛋白的水平及其臨床意義[J];上海醫(yī)學(xué)檢驗(yàn)雜志;1999年05期

5 何韜;王海杰;譚玉珍;;自噬在細(xì)胞存活和死亡中的作用[J];生理科學(xué)進(jìn)展;2008年01期

6 舒茂國(guó),郭樹忠,韓巖,楊力,張琳西,賈平,夏文森;機(jī)械應(yīng)力作用皮膚成纖維細(xì)胞增殖的相關(guān)研究[J];中國(guó)實(shí)用美容整形外科雜志;2004年01期

7 喬騁,陳紹宗,許龍順;負(fù)壓促進(jìn)豬深Ⅱ度燒傷創(chuàng)面愈合的實(shí)驗(yàn)研究[J];中國(guó)燒傷創(chuàng)瘍雜志;2000年02期

8 張自鵬;陳紹宗;李學(xué)擁;李金清;呂小星;雷戰(zhàn)軍;王曉琳;;封閉負(fù)壓引流聯(lián)合局部給氧治療兔耳缺血性創(chuàng)面的實(shí)驗(yàn)研究[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2011年10期

9 劉海波;陳紹宗;李學(xué)擁;李金清;雷戰(zhàn)軍;蔡樂;徐曉麗;;VAC技術(shù)治療兔耳缺血性創(chuàng)面的實(shí)驗(yàn)研究[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2012年11期

10 于風(fēng)旭;張英;凌生林;石應(yīng)康;廖斌;吳江;;不同剪切應(yīng)力對(duì)內(nèi)皮細(xì)胞表面SR-B1 mRNA表達(dá)影響的實(shí)驗(yàn)研究[J];生物醫(yī)學(xué)工程學(xué)雜志;2011年01期

本文編號(hào)：1590208