本文選題：燒沖復(fù)合傷 + 沖擊傷�。� 參考：《南開(kāi)大學(xué)》2013年博士論文

【摘要】：目的：燒沖復(fù)合傷是軍事沖突、恐怖襲擊、交通事故等突發(fā)事件中最常見(jiàn)的損傷類(lèi)型之一。燒沖復(fù)合傷后肺臟損傷導(dǎo)致即時(shí)死亡的發(fā)生率高達(dá)47%。急性肺損傷(ALI)和急性呼吸窘迫綜合癥(ARDS)是燒沖復(fù)合傷早期死亡的主要原因之一,燒沖復(fù)合傷后即刻引起低血容量性休克與炎癥級(jí)聯(lián)反應(yīng)。大量研究結(jié)果顯示在ALI/ARDS的發(fā)病過(guò)程中,細(xì)胞因子與中性粒細(xì)胞發(fā)揮了關(guān)鍵性的作用。IL-33是新近發(fā)現(xiàn)的一種細(xì)胞因子,屬于IL-1家族。IL-33mRNA表達(dá)于人類(lèi)與小鼠的多個(gè)器官和不同類(lèi)型的細(xì)胞,在蛋白水平,IL-33主要表達(dá)在上皮細(xì)胞、內(nèi)皮細(xì)胞和纖維細(xì)胞。IL-33能夠促進(jìn)中性粒細(xì)胞趨化至感染部位減輕膿毒癥。然而,在燒沖復(fù)合傷后肺組織IL-33與中性粒細(xì)胞是否存在這種機(jī)制仍不清楚。因此,我們建立重度燒沖復(fù)合傷動(dòng)物模型,探討在燒沖復(fù)合傷動(dòng)物模型與燒沖復(fù)合傷患者中IL-33是否參與肺損傷。 方法：利用野生型C57BL/6小鼠與IL-33轉(zhuǎn)基因小鼠分別建立燒沖復(fù)合傷、沖擊傷、燒傷動(dòng)物模型,小鼠隨機(jī)分為正常對(duì)照組,燒傷組,沖擊傷組,燒沖復(fù)合傷組。通過(guò)免疫組織化學(xué)法分別檢測(cè)肺組織髓過(guò)氧化物酶(myeloperoxidase,MPO)、白介素-33(IL-33)、G蛋白偶聯(lián)受體-2(GRK2)、趨化因子CXCR2的表達(dá)。透射電鏡(transmission electron microscopy)觀察肺組織超微結(jié)構(gòu)的變化。real-time RT-PCR檢測(cè)IL-33mRNA與GRK2mRNA的表達(dá)。酶聯(lián)免疫吸附實(shí)驗(yàn)(ELISA)檢測(cè)血漿IL-6與TNF-a濃度。Micro-CT觀察肺組織的變化。同時(shí),通過(guò)免疫組織化學(xué)法檢測(cè)燒沖復(fù)合傷患者傷后24小時(shí)肺組織MPO、IL-33的表達(dá)。酶聯(lián)免疫吸附實(shí)驗(yàn)檢測(cè)燒沖復(fù)合傷患者血漿IL-6與TNF-a濃度。胸部X-ray觀察肺組織的變化。 結(jié)果： 1.我們以5g8701炸藥為爆炸源,距離爆炸源43cm,53cm,63cm分別建立沖擊傷動(dòng)物模型,壓力值分別為200Kpa,94Kpa,88.3Kpa,根據(jù)沖擊傷診斷標(biāo)準(zhǔn)確定中度沖擊傷距離為53cm。 2.我們的動(dòng)物模型表明燒沖復(fù)合傷后肺組織嚴(yán)重?fù)p傷。與正常對(duì)照組小鼠肺組織肉眼觀相比,燒傷組小鼠肺組織輕度充血；沖擊傷組小鼠肺組織中度充血,可見(jiàn)約3處點(diǎn)灶狀出血；復(fù)合傷組小鼠肺組織明顯充血,可見(jiàn)約5處斑點(diǎn)狀出血；燒沖復(fù)合傷組小鼠傷后24小時(shí)與正常對(duì)照組、燒傷組、沖擊傷組相比,肺組織濕干比重、出血面積明顯增大。 3.與各實(shí)驗(yàn)組相比,在致傷后24小時(shí)燒沖復(fù)合傷小鼠的呼吸頻率(Rr)、潮氣量(Tv)、每分通氣量(Mv)、最大呼氣流量(PEF)明顯降低降低,然而呼氣時(shí)間(Te)與吸氣時(shí)間(Ti)顯著延長(zhǎng)。 4.傷后24小時(shí)micro-CT檢查顯示：正常對(duì)照組與燒傷組小鼠肺組織未見(jiàn)病變；沖擊傷組小鼠左側(cè)肺組織可見(jiàn)輕度的毛玻璃樣變區(qū)域；燒沖復(fù)合傷組小鼠左側(cè)肺組織可見(jiàn)中度毛玻璃樣變區(qū)域。 5.燒沖復(fù)合傷野生型C57BL/6小鼠傷后24小時(shí)肺組織髓過(guò)氧化物酶(MPO)、白介素一33(IL-33)、G-蛋白偶聯(lián)受體-2(GRK2)表達(dá)明顯增強(qiáng)。與燒沖復(fù)合傷后IL-33轉(zhuǎn)基因小鼠傷后24小時(shí)相比,燒沖復(fù)合傷野生型C57BL/6小鼠趨化因子(CXCR2)明顯減少；與各實(shí)驗(yàn)組相比,燒沖復(fù)合傷組小鼠血漿IL-6與TNF-α濃度均明顯升高。 6.在燒傷、沖擊傷、燒沖復(fù)合傷小鼠肺組織中IL-33的陽(yáng)性細(xì)胞數(shù)與GRK2陽(yáng)性細(xì)胞數(shù)、MPO陽(yáng)性細(xì)胞數(shù)均有顯著的正相關(guān)(r=0.65,p=0.042；r=0.638, p=0.0047；r=0.764,p=0.001)(r=0.716,p=0.02；r=0.661,p=0.0037；r=0.755, p=0.012),與Mv呈負(fù)相關(guān)(r=-0.677,p=0.031;r=-0.68,p=0.03;r=-0.816,p=0.004). 7.燒沖復(fù)合傷尸檢見(jiàn)傷側(cè)肺組織表面可見(jiàn)散在的斑片狀出血,HE染色觀察見(jiàn)大面積肺泡內(nèi)出血。肺組織標(biāo)本免疫組化染色見(jiàn)IL-33陽(yáng)性細(xì)胞數(shù)高表達(dá)區(qū)域MPO陽(yáng)性細(xì)胞數(shù)也高表達(dá)。 8.燒沖復(fù)合傷患者傷側(cè)肺組織呈中度毛玻璃樣變。 結(jié)論： 1.成功的建立了一種燒沖復(fù)合傷小鼠動(dòng)物模型,具體參數(shù)為：野生型C57BL/6小鼠距離爆炸源53cm,壓力峰值94Kpa致中度沖擊傷,即刻將中度沖擊傷小鼠背部脫毛區(qū)置入90℃沸水中持續(xù)9s,造成小鼠背部25%TBSAⅢ度燙傷。 2. IL-33通過(guò)激活GRK2通路阻斷趨化因子CXCR2下調(diào)促進(jìn)中性粒細(xì)胞遷移參與燒沖復(fù)合傷小鼠肺損傷。 3.高表達(dá)的IL-33參與燒沖復(fù)合傷患者肺損傷。
[Abstract]:Objective: burn and impulse compound injury is one of the most common types of injuries in military, terrorist, and traffic accidents. The incidence of immediate death caused by lung injury after burn and impulse compound injury is as high as 47%. acute lung injury (ALI) and acute respiratory distress syndrome (ARDS), one of the main causes of early death of burning and punching complex injuries. A number of results show that cytokine and neutrophils play a key role in the pathogenesis of ALI/ARDS, and.IL-33 is a newly discovered cytokine in the pathogenesis of ALI/ARDS, which belongs to the multiple organs and different types of human and mice in the family.IL-33mRNA. Type of cells, at protein level, IL-33 is mainly expressed in epithelial cells, endothelial cells and fibrous cell.IL-33 to promote neutrophil chemotaxis to sepsis. However, it is not clear whether the mechanism of IL-33 and neutrophils exists in the lung tissue after burn and blast injury. Objective to investigate whether IL-33 is involved in lung injury in patients with combined burn blast combined injury and combined burn blast injury.
Methods: the wild type C57BL/6 mice and IL-33 transgenic mice were used to establish the burning and punching complex injury, the impact injury and the burn animal model. The mice were randomly divided into the normal control group, the burn group, the impact injury group and the burn and flush compound injury group. The pulmonary myeloperoxidase (myeloperoxidase, MPO) and interleukin -33 (IL-33) were detected by immunohistochemistry. G protein coupling receptor -2 (GRK2), the expression of chemokine CXCR2. Transmission electron microscopy (transmission electron microscopy) observed changes in the ultrastructure of the lung tissue and the expression of.Real-time RT-PCR detection IL-33mRNA and GRK2mRNA. The expression of MPO and IL-33 in the lung tissue was detected by the method of immunohistochemistry for 24 hours after injury. The plasma concentration of IL-6 and TNF-a was detected by enzyme linked immunosorbent assay (ELISA). The changes of lung tissue were observed by chest X-ray.
Result錛，

本文編號(hào)：1923605