爆炸致家兔急性肺损伤及血必净保护作用的实验研究

发布时间：2018-03-22 10:21

  本文选题：急性肺损伤　切入点：血必净　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的观察血必净对爆炸致家兔急性肺损伤(ALI)的保护作用,并研究探讨其作用机制。方法健康清洁级家兔32只,采用简单随机化分组方法分成4组:正常对照(NS)组、爆炸模型(ALI)组和低剂量血必净(L-XBJ)组、高剂量血必净(H-XBJ)组,每组8只,由指定专业人员以工业炸药为爆炸源,爆炸致伤家兔,爆炸后迅速实施简单的急救措施,观察并记录家兔的一般情况,精神状态,应激反应等。建立ALI动物模型后,L-XBJ组、H-XBJ组分别注射10、20 ml/kg血必净溶液,NS组、ALI组注射生理盐水。4 h采集静脉血,24 h实验结束时采集支气管肺泡灌洗液(BALF)和部分肺组织标本,Western blot法检测肺组织MMP-9蛋白和NF-κB蛋白的表达水平,反转录PCR法测定肺组织MMP-9 m RNA和NF-κB m RNA的表达水平,ELISA法测定BALF及血清中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、白介素-10(IL-10)的含量,并测定肺湿干重比(W/D),光镜下观察肺组织切片病理学改变。结果1、各组家兔的一般情况:实验前,各组家兔均发育良好,自由饮食、饮水,呼吸频率、活动能力正常。爆炸致伤后,ALI组、L-XBJ组、H-XBJ组家兔呼吸急促,心率加快,躁动,口唇、耳鼻部等皮肤黏膜呈发绀表现,口鼻腔黏膜充血伴泡沫状分泌物,双肺部干、湿Up音明显,伴呼吸困难、窘迫等症状,少数家兔短暂性昏迷。2、各组家兔肺组织大体表现:NS组家兔肺组织表面为均匀粉红色,包膜完整光滑,富有弹性,无破损,切面细腻紧致,无蛋白水肿液溢出。ALI组肺组织表面颜色暗红,体积显著增加,包膜下充血,肺表面可见多发片状出血灶,切面有泡沫状蛋白水肿液溢出。L-XBJ组、H-XBJ组肺组织表面颜色暗红,包膜下充血程度较ALI组轻,体积也较ALI组小。3、Western blot法测定肺组织MMP-9蛋白:NS组仅少量表达,ALI组明显高表达,随血必净剂量增加,L-XBJ组、H-XBJ组的蛋白表达逐渐降低,以H-XBJ组降低最为显著。4、RT-PCR测定肺组织MMP-9 m RNA的表达水平:经方差分析,MMP-9 m RNA的表达水平差异有统计学意义(P0.01)。与NS组比较,ALI组明显高表达(P0.01),随血必净剂量增加,L-XBJ组、H-XBJ组的表达水平逐渐降低,以H-XBJ组降低最为显著(P0.01)。5、Western blot法测定肺组织NF-κB蛋白:NS组仅少量表达,ALI组明显高表达,随血必净剂量增加,L-XBJ组、H-XBJ组的蛋白表达逐渐降低,以H-XBJ组降低最为显著。6、RT-PCR测定肺组织NF-κB m RNA的表达水平:经方差分析,NF-κB m RNA的表达水平差异有统计学意义(P0.01)。与NS组比较,ALI组明显高表达(P0.01),随血必净剂量增加,L-XBJ组、H-XBJ组的表达水平逐渐降低,以H-XBJ组降低最为显著(P0.01)。7、各组家兔肺组织湿干重比(W/D)含量比较:ALI组、L-XBJ组、H-XBJ组含量均较NS组升高,L-XBJ组、H-XBJ组含量较ALI组降低,以H-XBJ组降低最为显著(P0.05)。8、各组家兔4h血清TNF-α、IL-6含量比较:与NS组比较,ALI组高表达(P0.05),经血必净干预后,L-XBJ组、H-XBJ组指标均较ALI组降低,以H-XBJ组降低最为显著(P0.05)。9、各组家兔4h血清IL-10含量比较:经血必净干预后,L-XBJ组、H-XBJ组均较ALI组升高,且H-XBJ组升高更显著(P0.05)。10、各组家兔肺泡灌洗液TNF-α、IL-6含量比较:与NS组比较,ALI组高表达(P0.05),随血必净剂量增加,L-XBJ组、H-XBJ组指标均较ALI组降低,以H-XBJ组降低最为显著(P0.05)。11、各组家兔肺泡灌洗液IL-10含量比较:经血必净干预后,L-XBJ组、H-XBJ组均较ALI组升高,且H-XBJ组升高更显著(P0.05)。12、各组家兔肺组织病理学改变:NS组为正常肺组织,组织结构清晰可见,肺泡壁完整,肺泡腔干净,无炎性细胞浸润,间质无充血水肿液渗出。ALI组肺泡壁断裂,肺泡腔和肺间质重度水肿,大量红细胞和炎性细胞渗出,偶见透明膜,肺泡间隔断裂、距离增宽,光镜下视野肺泡大小不均,部分视野可见肺不张及肺气肿。L-XBJ组、H-XBJ组肺泡腔和肺间质红细胞、蛋白水肿液、炎性细胞渗出较ALI组减轻,以H-XBJ组减轻最为明显。结论1、爆炸致伤后家兔一般情况、湿干重比联合病理学变化,提示爆炸致家兔ALI模型制作成功。2、血必净可抑制肺部的失控性炎症反应,降低肺内蛋白渗出,作用机制可能跟阻断NF-κB介导的炎症反应通路,降低MMP-9的表达,以及下调致炎因子TNF-α、IL-6水平,上调抗炎因子IL-10水平有关,从而减轻肺损伤,保护肺组织。
[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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