本文選題：急性肺損傷　切入點(diǎn)：血必凈　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的觀察血必凈對爆炸致家兔急性肺損傷(ALI)的保護(hù)作用,并研究探討其作用機(jī)制。方法健康清潔級家兔32只,采用簡單隨機(jī)化分組方法分成4組:正常對照(NS)組、爆炸模型(ALI)組和低劑量血必凈(L-XBJ)組、高劑量血必凈(H-XBJ)組,每組8只,由指定專業(yè)人員以工業(yè)炸藥為爆炸源,爆炸致傷家兔,爆炸后迅速實(shí)施簡單的急救措施,觀察并記錄家兔的一般情況,精神狀態(tài),應(yīng)激反應(yīng)等。建立ALI動物模型后,L-XBJ組、H-XBJ組分別注射10、20 ml/kg血必凈溶液,NS組、ALI組注射生理鹽水。4 h采集靜脈血,24 h實(shí)驗(yàn)結(jié)束時(shí)采集支氣管肺泡灌洗液(BALF)和部分肺組織標(biāo)本,Western blot法檢測肺組織MMP-9蛋白和NF-κB蛋白的表達(dá)水平,反轉(zhuǎn)錄PCR法測定肺組織MMP-9 m RNA和NF-κB m RNA的表達(dá)水平,ELISA法測定BALF及血清中腫瘤壞死因子-α(TNF-α)、白介素-6(IL-6)、白介素-10(IL-10)的含量,并測定肺濕干重比(W/D),光鏡下觀察肺組織切片病理學(xué)改變。結(jié)果1、各組家兔的一般情況:實(shí)驗(yàn)前,各組家兔均發(fā)育良好,自由飲食、飲水,呼吸頻率、活動能力正常。爆炸致傷后,ALI組、L-XBJ組、H-XBJ組家兔呼吸急促,心率加快,躁動,口唇、耳鼻部等皮膚黏膜呈發(fā)紺表現(xiàn),口鼻腔黏膜充血伴泡沫狀分泌物,雙肺部干、濕Up音明顯,伴呼吸困難、窘迫等癥狀,少數(shù)家兔短暫性昏迷。2、各組家兔肺組織大體表現(xiàn):NS組家兔肺組織表面為均勻粉紅色,包膜完整光滑,富有彈性,無破損,切面細(xì)膩緊致,無蛋白水腫液溢出。ALI組肺組織表面顏色暗紅,體積顯著增加,包膜下充血,肺表面可見多發(fā)片狀出血灶,切面有泡沫狀蛋白水腫液溢出。L-XBJ組、H-XBJ組肺組織表面顏色暗紅,包膜下充血程度較ALI組輕,體積也較ALI組小。3、Western blot法測定肺組織MMP-9蛋白:NS組僅少量表達(dá),ALI組明顯高表達(dá),隨血必凈劑量增加,L-XBJ組、H-XBJ組的蛋白表達(dá)逐漸降低,以H-XBJ組降低最為顯著。4、RT-PCR測定肺組織MMP-9 m RNA的表達(dá)水平:經(jīng)方差分析,MMP-9 m RNA的表達(dá)水平差異有統(tǒng)計(jì)學(xué)意義(P0.01)。與NS組比較,ALI組明顯高表達(dá)(P0.01),隨血必凈劑量增加,L-XBJ組、H-XBJ組的表達(dá)水平逐漸降低,以H-XBJ組降低最為顯著(P0.01)。5、Western blot法測定肺組織NF-κB蛋白:NS組僅少量表達(dá),ALI組明顯高表達(dá),隨血必凈劑量增加,L-XBJ組、H-XBJ組的蛋白表達(dá)逐漸降低,以H-XBJ組降低最為顯著。6、RT-PCR測定肺組織NF-κB m RNA的表達(dá)水平:經(jīng)方差分析,NF-κB m RNA的表達(dá)水平差異有統(tǒng)計(jì)學(xué)意義(P0.01)。與NS組比較,ALI組明顯高表達(dá)(P0.01),隨血必凈劑量增加,L-XBJ組、H-XBJ組的表達(dá)水平逐漸降低,以H-XBJ組降低最為顯著(P0.01)。7、各組家兔肺組織濕干重比(W/D)含量比較:ALI組、L-XBJ組、H-XBJ組含量均較NS組升高,L-XBJ組、H-XBJ組含量較ALI組降低,以H-XBJ組降低最為顯著(P0.05)。8、各組家兔4h血清TNF-α、IL-6含量比較:與NS組比較,ALI組高表達(dá)(P0.05),經(jīng)血必凈干預(yù)后,L-XBJ組、H-XBJ組指標(biāo)均較ALI組降低,以H-XBJ組降低最為顯著(P0.05)。9、各組家兔4h血清IL-10含量比較:經(jīng)血必凈干預(yù)后,L-XBJ組、H-XBJ組均較ALI組升高,且H-XBJ組升高更顯著(P0.05)。10、各組家兔肺泡灌洗液TNF-α、IL-6含量比較:與NS組比較,ALI組高表達(dá)(P0.05),隨血必凈劑量增加,L-XBJ組、H-XBJ組指標(biāo)均較ALI組降低,以H-XBJ組降低最為顯著(P0.05)。11、各組家兔肺泡灌洗液IL-10含量比較:經(jīng)血必凈干預(yù)后,L-XBJ組、H-XBJ組均較ALI組升高,且H-XBJ組升高更顯著(P0.05)。12、各組家兔肺組織病理學(xué)改變:NS組為正常肺組織,組織結(jié)構(gòu)清晰可見,肺泡壁完整,肺泡腔干凈,無炎性細(xì)胞浸潤,間質(zhì)無充血水腫液滲出。ALI組肺泡壁斷裂,肺泡腔和肺間質(zhì)重度水腫,大量紅細(xì)胞和炎性細(xì)胞滲出,偶見透明膜,肺泡間隔斷裂、距離增寬,光鏡下視野肺泡大小不均,部分視野可見肺不張及肺氣腫。L-XBJ組、H-XBJ組肺泡腔和肺間質(zhì)紅細(xì)胞、蛋白水腫液、炎性細(xì)胞滲出較ALI組減輕,以H-XBJ組減輕最為明顯。結(jié)論1、爆炸致傷后家兔一般情況、濕干重比聯(lián)合病理學(xué)變化,提示爆炸致家兔ALI模型制作成功。2、血必凈可抑制肺部的失控性炎癥反應(yīng),降低肺內(nèi)蛋白滲出,作用機(jī)制可能跟阻斷NF-κB介導(dǎo)的炎癥反應(yīng)通路,降低MMP-9的表達(dá),以及下調(diào)致炎因子TNF-α、IL-6水平,上調(diào)抗炎因子IL-10水平有關(guān),從而減輕肺損傷,保護(hù)肺組織。
[Abstract]:Objective To observe the effect of Xuebijing on blast induced acute lung injury in rabbits (ALI) of the protective effect, and explore its mechanism. Methods 32 clean rabbits, using simple random grouping method is divided into 4 groups: normal control group (NS), explosion model (ALI) group and low dose of Xuebijing group (L-XBJ), high dose of Xuebijing (H-XBJ) group, 8 rats in each group, designated by the professionals in industrial explosives for explosive injury in rabbits induced by explosion, explosion after the rapid implementation of simple emergency measures, observe and record the general situation, in mental state, stress reaction. To establish an animal model of ALI, L-XBJ group, H-XBJ group respectively. Injection of 10,20 ml/kg NS group, ALI Xuebijing solution, saline.4 h collected venous blood at the end of the 24 h experiment collected bronchoalveolar lavage fluid (BALF) and lung tissue samples, the expression of Western blot in lung tissues were detected MMP-9 protein and NF- kappa B protein The level of expression of RNA and m in lung tissue was determined MMP-9 NF- kappa B m RNA reverse transcription PCR method, determination of tumor necrosis factor alpha BALF in serum and ELISA (TNF- alpha), interleukin -6 (IL-6), interleukin -10 (IL-10) content, and the determination of lung wet to dry weight ratio (W/D under the light microscope), lung tissue pathology. Results 1 general condition of rabbits: before the experiment, all rabbits were well developed, free food, drinking water, breathing frequency, normal activity. After blast injury, ALI group, L-XBJ group, H-XBJ group of rabbits with shortness of breath, heart rate fast. Restlessness, lips, and other skin ear and nose mucosa was cyanosis, nasal mucosa hyperemia with foamy mouth secretions, double lung dry, wet rales, Up, with dyspnea, distress and other symptoms, a rabbit transient coma.2, lung tissue of rabbits in group NS: rabbit lung tissue surface is uniformly pink. Complete capsule is smooth, rich Elastic, no damage, no section of exquisite compact, the color of the surface protein of edema fluid overflow in lung tissue in group.ALI dishes, volume increased significantly, subcapsular congestion, lung surface visible multiple patchy hemorrhage, a section of foam protein edema fluid overflow in.L-XBJ group, H-XBJ group, lung tissue surface color dark red, subcapsular hyperemia compared with the ALI group, the volume is smaller than the group ALI.3, Western blot MMP-9 protein in lung tissue: in NS group, only a small amount of expression, ALI expression was significantly higher, with the increase of the dose of Xuebijing, group L-XBJ, group the expression of H-XBJ protein decreased gradually in H-XBJ group was reduced significantly.4, RT-PCR expression level determination of lung MMP-9 m RNA: through analysis of variance, there was significant difference in the expression level of MMP-9 m RNA (P0.01). Compared with group NS, group ALI expression was significantly higher (P0.01), with the increase of the dose of Xuebijing injection, L-XBJ group, the expression level of H-XBJ group decreased in H-XBJ group 闄嶄綆鏈，

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