本文選題：急性肺挫傷　切入點(diǎn)：TLR-9受體　出處：《貴州醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:Toll受體9(Toll-like receptors-9,TLR-9)在胸部創(chuàng)傷所致大鼠急性肺挫傷中表達(dá)的意義。方法:選取2016年4月~5月購(gòu)自貴州醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)中心的40只健康、雄性SD(Sprague-Dawley,SD)大鼠作為研究對(duì)象。利用小型生物撞擊機(jī)(由第三軍醫(yī)大學(xué)予以提供),在500kpa下對(duì)大鼠胸部進(jìn)行撞擊,形成重度肺挫傷。參照AIS-90標(biāo)準(zhǔn),對(duì)受傷后大鼠進(jìn)行評(píng)分,大鼠受傷后會(huì)出現(xiàn)頻繁、深沉的呼吸,較之前更明顯,呼吸急促、口唇發(fā)鉗、蘇醒延遲、精神萎靡、食欲減退、活動(dòng)減少。上述提示:代謝性呼吸增快在大鼠身上已經(jīng)出現(xiàn),目的在于適應(yīng)氧供。因此,采用500kpa撞擊較好的模擬了大鼠急性肺挫傷。隨機(jī)分為5組,每組8只:正常對(duì)照組;胸部創(chuàng)傷致急性肺挫傷模擬組(500kpa)撞擊后5,24,48,72小時(shí),于股動(dòng)脈放血處死。取大鼠血清及肺組織。HE染色觀察大鼠肺組織病理學(xué)改變、肺組織濕/干質(zhì)量比、采用免疫組織化學(xué)法、Elisa法檢測(cè)血清及肺泡灌洗液中TLR-9受體的蛋白表達(dá)水平、予RT-PCR(Reverse transcription-PCR,RT-PCR)法檢測(cè)肺組織TLR-9、IL-6(Interleukin-6,IL-6)的mRNA表達(dá)水平;Western blot法檢測(cè)肺組織TLR-9的蛋白水平。結(jié)果:肺組織病理學(xué)檢查顯示:與空白對(duì)照組比較,胸部創(chuàng)傷組大鼠肺水腫、肺組織充血、中性粒細(xì)胞浸潤(rùn)、炎性滲出較明顯。免疫組織化學(xué)法回示:胸部創(chuàng)傷組大鼠光鏡下顯色呈明顯棕黃色,空白對(duì)照組大鼠光鏡下未見黃色,其中以胸部創(chuàng)傷48小時(shí)組大鼠染色最為明顯。胸部創(chuàng)傷組大鼠肺組織濕/干質(zhì)量比:胸部創(chuàng)傷組大鼠與空白對(duì)照組大鼠相比,胸部創(chuàng)傷組大鼠肺組織濕/干質(zhì)量比明顯低于空白對(duì)照組。ELISA:血清及肺泡灌洗液中TLR-9、IL-6表達(dá)水平,胸部創(chuàng)傷組高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。RT-PCR回示:胸部創(chuàng)傷組肺組織中TLR-9 mRNA、IL-6 mRNA的表達(dá)較空白對(duì)照組升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。Western blot回示:胸部創(chuàng)傷組肺組織中TLR-9蛋白水平較空白對(duì)照組升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:大鼠創(chuàng)傷后TLR-9表達(dá)的增高,提示TLR-9參加了肺挫傷炎癥反應(yīng)的過程。
[Abstract]:Objective: to investigate the expression of Toll-like receptor 9 (TLR-9) in acute lung contusion induced by thoracic trauma in rats. Methods: from April 2016 to May, 40 healthy rats were selected from the Animal Experimental Center of Guizhou Medical University. Sprague-Dawley SD rats were used as the study subjects. Using a small biological impact machine (provided by the third military Medical University), the chest of the rats was impacted at 500kpa, resulting in severe lung contusion. According to AIS-90 criteria, the injured rats were graded. There are frequent, deep breaths in rats after injury, more pronounced than before, shortness of breath, lip forceps, delayed recovery, depression of mind, loss of appetite, and decreased activity. The aim was to adapt to oxygen supply. Therefore, acute lung contusion was simulated by 500kpa impact in rats. The rats were randomly divided into 5 groups: normal control group (n = 8), acute lung contusion simulation group induced by chest trauma (n = 500 kpa) and control group (n = 8). The pathological changes of lung tissue were observed by HE staining in serum and lung tissue. The wet / dry weight ratio of lung tissue was measured. The expression of TLR-9 receptor protein in serum and alveolar lavage fluid was detected by immunohistochemistry. The expression level of mRNA in lung tissue was detected by RT-PCR(Reverse transcription-PCRD-PCRRT) and the protein level of TLR-9 in lung tissue was detected by Western blot. Results: compared with the control group, pulmonary edema and congestive lung tissue were detected by RT-PCR(Reverse transcription-PCRRT. Neutrophil infiltration and inflammatory exudation were obvious. The results of immunohistochemistry showed that the rats of thoracic trauma group showed obvious brownish yellow under light microscope, but no yellow was found in blank control group under light microscope. The wet / dry weight ratio of lung tissue in the thoracic trauma group was higher than that in the blank control group, and the wet / dry weight ratio of lung tissue in the thoracic trauma group was higher than that in the blank control group. The wet / dry weight ratio of lung tissue in the thoracic trauma group was significantly lower than that in the blank control group. The expression of TLR-9P IL-6 in serum and alveolar lavage fluid in the thoracic trauma group was significantly lower than that in the control group, and that in the chest trauma group was higher than that in the control group. The expression of TLR-9 mRNA-IL-6 mRNA in lung tissue of thoracic trauma group was higher than that of blank control group, and the level of TLR-9 protein in lung tissue of thoracic trauma group was higher than that of blank control group. Conclusion: the increase of TLR-9 expression in rats after trauma suggests that TLR-9 participates in the process of pulmonary contusion inflammation.
【學(xué)位授予單位】：貴州醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R655.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 吳雪花;王昌鋒;彭金娥;;嚴(yán)重肺挫傷早期行機(jī)械通氣治療的療效觀察[J];臨床肺科雜志;2017年03期

2 金平飛;賀淼;;TLR/NF-κB信號(hào)通路與肺部疾病[J];生命的化學(xué);2016年06期

3 楊雪姣;劉萬成;祝元鋒;楊永軍;范仕郡;李彥;;CpG-c41對(duì)TLR9信號(hào)通路的干擾作用及其機(jī)制探究[J];免疫學(xué)雜志;2016年12期

4 周楊;董佑紅;;TLR在乳腺癌中的作用及其治療進(jìn)展[J];生命的化學(xué);2016年04期

5 劉沙沙;劉冰熔;;TLR4、TLR9與結(jié)直腸癌關(guān)系的研究進(jìn)展[J];胃腸病學(xué)和肝病學(xué)雜志;2015年03期

6 胥靜;丁力;張俊平;;Toll樣受體和其他分子識(shí)別受體在固有免疫中的相互作用[J];藥學(xué)實(shí)踐雜志;2014年05期

7 喬良;劉志;;急診肺炎患者發(fā)生急性肺損傷/急性呼吸窘迫綜合征危險(xiǎn)因素分析[J];生物醫(yī)學(xué)工程與臨床;2014年05期

8 馮健華;;TLR9信號(hào)通路參與急性肺損傷的作用機(jī)制研究[J];國(guó)際醫(yī)藥衛(wèi)生導(dǎo)報(bào);2014年11期

9 劉新華;;急性肺損傷/急性呼吸窘迫綜合征發(fā)病機(jī)制的研究進(jìn)展[J];中國(guó)衛(wèi)生產(chǎn)業(yè);2014年01期

10 施卉;任成山;;急性肺損傷/急性呼吸窘迫綜合征基礎(chǔ)及臨床研究進(jìn)展[J];中華肺部疾病雜志(電子版);2013年04期

相關(guān)會(huì)議論文 前1條

1 張?jiān)姾?;急性肺損傷的研究進(jìn)展[A];中華醫(yī)學(xué)會(huì)第二十次全國(guó)麻醉學(xué)術(shù)年會(huì)論文匯編[C];2012年

相關(guān)碩士學(xué)位論文 前2條

1 謝輝;降鈣素基因相關(guān)肽對(duì)ALI時(shí)小鼠巨噬細(xì)胞替代激活的影響[D];中南大學(xué);2013年

2 陸駿灝;嚴(yán)重肺挫傷患者血中VEGF、vWF及IL-10的動(dòng)態(tài)變化及其意義[D];蘇州大學(xué);2009年

，

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