【摘要】：目的：通過研究低氧誘導(dǎo)因子-1α (Hypoxia inducing factor-1α)在銅綠假單胞菌肺炎模型小鼠肺組織中的表達(dá)情況，以及探討HIF-1α與TNF-α、IL-8之間的相關(guān)性，推測HIF-1α在銅綠假單胞菌肺炎中的可能作用，以期為銅綠假單胞菌肺炎的治療尋找到新思路。方法：1.將鼠齡6~8周、體重25~30g的清潔級健康雄性昆明小鼠120只按隨機(jī)原則分成肺部感染組（PA組：60只）和正常對照組（生理鹽水組：60只）。PA組：氯胺酮麻醉小鼠后固定，常規(guī)消毒鋪巾，嚴(yán)格無菌操作，切開頸部皮膚，鈍性分離至皮下組織，暴露上段氣管，經(jīng)氣管內(nèi)穿刺注入銅綠假單胞菌（PA）菌液（2個(gè)麥?zhǔn)蠁挝粷舛取?.03ml）建立小鼠銅綠假單胞菌肺炎模型；正常對照組：操作同PA組，經(jīng)氣管內(nèi)穿刺注入無菌生理鹽水（0.03ml）作為對照，兩組均于造模成功后第3、9、24小時(shí)隨機(jī)分別抽取20只小鼠，予以留取標(biāo)本后處死。2.收集各組不同時(shí)間點(diǎn)處死的小鼠血液，離心分離血清后，以酶聯(lián)免疫吸附試驗(yàn)（enzyme-linked immunosorbnent assay，ELISA）檢測血清中TNF-α、IL-8的濃度；取右肺經(jīng)10%多聚甲醛固定后，切片行HE染色比較炎癥的范圍及程度，并行免疫組化染色檢測肺組織中HIF-1α蛋白的表達(dá)水平；同時(shí)留取左側(cè)肺組織行肺組織勻漿細(xì)菌培養(yǎng)及菌落計(jì)數(shù)。3.應(yīng)用SPSS17.0軟件對數(shù)據(jù)進(jìn)行統(tǒng)計(jì)分析。結(jié)果：1、PA感染組小鼠肺組織病理改變在24小時(shí)最為嚴(yán)重，炎癥范圍廣，程度重，3、9小時(shí)與之相比均較輕，，但均有明顯的炎癥改變；而對照組的肺泡及肺間質(zhì)結(jié)構(gòu)基本正常，無明顯出血及炎細(xì)胞浸潤。并且PA感染組3、9、24小時(shí)肺組織勻漿均培養(yǎng)出銅綠假單胞菌，菌落計(jì)數(shù)均105CFU/g，而對照組肺組織勻漿各時(shí)間點(diǎn)均未培養(yǎng)出細(xì)菌。2、PA感染組3、9、24小時(shí)小鼠肺組織中HIF-1α蛋白的表達(dá)均有明顯的增高，其中細(xì)菌接種3小時(shí)后HIF-1α的表達(dá)就有升高，9小時(shí)繼續(xù)升高，24小時(shí)最高。而對照組HIF-1α各時(shí)間點(diǎn)均未見明顯表達(dá)。3、PA感染組3、9、24小時(shí)小鼠血清中TNF-α、IL-8的水平較對照組均有明顯的增高，差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。其中TNF-α、IL-8的水平均在3小時(shí)開始上升，9小時(shí)繼續(xù)升高，24小時(shí)最高。4、HIF-1α蛋白的表達(dá)與TNF-α、IL-8的濃度呈正相關(guān)（相關(guān)系數(shù)分別為0.827和0.974）。結(jié)論：1. HIF-1α參與了銅綠假單胞菌肺炎的炎癥過程，且炎癥愈重表達(dá)愈明顯。2. HIF-1α是炎癥細(xì)胞因子網(wǎng)絡(luò)中的重要一環(huán)，它與TNF-α、IL-8等相互作用，促進(jìn)炎癥細(xì)胞合成及釋放多種炎癥因子，在銅綠假單胞菌肺炎的發(fā)生發(fā)展中起了重要作用。
[Abstract]:Objective: to study the expression of hypoxia inducible factor-1 偽 (Hypoxia inducing factor-1 偽 in lung tissue of mice with Pseudomonas aeruginosa pneumonia and to explore the correlation between HIF-1 偽 and TNF- 偽, IL-8. The possible role of HIF-1 偽 in Pseudomonas aeruginosa pneumonia was speculated in order to find a new idea for the treatment of Pseudomonas aeruginosa pneumonia. Method 1: 1. A total of 120 healthy male Kunming mice were randomly divided into two groups: lung infection group (PA group: 60 mice) and normal control group (normal saline group: 60). PA mice). Routine disinfecting and spreading towel, strict aseptic operation, incision of neck skin, blunt separation to subcutaneous tissue, exposure of upper trachea, puncture and injection of Pseudomonas aeruginosa (PA) solution (2 McClone unit concentrations) in trachea through trachea. 0.03ml) to establish the model of Pseudomonas aeruginosa pneumonia in mice. Normal control group: in the same operation as PA group, 20 mice were randomly selected at 24 hours after successful modeling and 20 mice were randomly selected from the two groups after puncture and injection of sterile saline (0.03ml) into the trachea. 2. 2. Blood samples were collected from mice killed at different time points. After centrifugation, the concentrations of TNF- 偽 and IL-8 in serum were detected by enzyme linked immunosorbent assay (enzyme-linked immunosorbnent assay,ELISA). After the right lung was fixed with 10% paraformaldehyde, HE staining was performed to compare the extent and degree of inflammation, and the expression of HIF-1 偽 protein in lung tissue was detected by immunohistochemical staining. At the same time, the left lung tissue was retained for the culture of bacteria and colony count of lung tissue homogenate. The data were analyzed by SPSS17.0 software. Results: 1 the pathological changes of lung tissue in PA infected mice were the most serious in 24 hours. The range of inflammation was wide and the degree of inflammation was severe. Compared with the control group, the pathological changes of lung tissue in the group of 3 ~ 9 hours were lighter, but there were obvious inflammatory changes. In the control group, the alveolar and interstitial structures were normal, no obvious bleeding and inflammatory cell infiltration. Pseudomonas aeruginosa was cultured in lung tissue homogenate of PA infection group at 24 hours, and colony counts were 105 CFU / g, but no bacteria were cultured in lung tissue homogenate of control group. The expression of HIF-1 偽 protein in lung tissue of 24 h mice was significantly increased. The expression of HIF-1 偽 was increased 3 hours after inoculation, and continued to increase at 9 h, and reached the highest level at 24 h. The levels of TNF- 偽 and IL-8 in the control group were significantly higher than those in the control group (P < 0. 05). The levels of TNF- 偽 and IL-8 began to rise at 3 hours, continued to increase at 9 hours, and reached the highest at 24 hours. The expression of HIF-1 偽 protein and TNF- 偽 in 24 hours were the highest. The concentration of IL-8 was positively correlated (correlation coefficient was 0.827 and 0.974, respectively). Conclusion 1. HIF-1 偽 was involved in the inflammatory process of Pseudomonas aeruginosa pneumonia, and the higher the inflammatory expression was, the more obvious. 2. HIF-1 偽 is an important link in the inflammatory cytokine network. It interacts with TNF- 偽, IL-8 and so on, promotes the synthesis and release of many inflammatory factors by inflammatory cells, and plays an important role in the occurrence and development of Pseudomonas aeruginosa pneumonia.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R563.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 方怡,冉丕鑫;重癥社區(qū)獲得性肺炎炎性機(jī)制研究進(jìn)展[J];國外醫(yī)學(xué).呼吸系統(tǒng)分冊;2005年01期

2 劉金波;方飛;易娟;易莉;;人牙髓組織缺氧耐受機(jī)制的初步研究[J];臨床口腔醫(yī)學(xué)雜志;2007年05期

3 許春梅,董衛(wèi)國,余保平,羅和生,于皆平;低氧誘導(dǎo)因子-1α和炎癥誘導(dǎo)酶在炎癥性腸病中的表達(dá)及意義[J];臨床消化病雜志;2005年03期

4 劉波;許春梅;;HIF-1α、COX-2在潰瘍性結(jié)腸炎中的表達(dá)及與疾病活動性的關(guān)系[J];臨床消化病雜志;2008年03期

5 敖啟林,黃磊,熊維寧,朱朋成,王偉,吳人亮,王迪潯;低氧誘導(dǎo)因子1α促進(jìn)大鼠肺泡巨噬細(xì)胞腫瘤壞死因子α的產(chǎn)生[J];中華結(jié)核和呼吸雜志;2004年06期

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