高原缺氧及布洛芬干预对大鼠细胞因子影响的机理研究

发布时间：2018-04-14 14:14

本文选题：缺氧模型 + 肺组织　；参考：《兰州大学》2015年硕士论文

【摘要】：快速进入高原环境时,人们常会因为缺氧,出现恶心、呕吐、乏力等一系列机体不适,严重者会出现急性高原疾病,如不及早治疗可能会造成患者死亡。然而,急性高原疾病的发病机制不明确,造成药物研发缓慢,使人们的生命健康安全遭受高原缺氧环境的威胁。因此,探究高原缺氧损伤机理,研发抗缺氧保护作用药物非常重要。本论文将建立急进高原实地缺氧大鼠模型,围绕缺氧对大鼠体内细胞因子的影响,探究高原环境对大鼠缺氧损伤的机制并考察抗炎药物布洛芬的潜在抗缺氧保护作用。课题组首先通过急进高原缺氧环境和高原低压低氧模拟舱模拟,建立大鼠缺氧模型,比较缺氧组和平原组大鼠的血气、生化、病理差异。结果表明,急进高原实地缺氧大鼠与高原低压低氧模拟舱模拟缺氧大鼠的血气、生化、病理指标变化趋势基本一致,其中,急性缺氧(模拟缺氧和实地缺氧)对大鼠的主要损伤器官为肺组织。结果还提示,在涉及血气、生化测定的高原缺氧研究中,应该对研究所用的缺氧模型做出选择,不同模型的结果差异可能较大。本结果为急进高原实地缺氧模型的建立提供实验依据。其次,采用Antibody Array技术比较高原缺氧组和平原组大鼠体内34种大鼠细胞因子的差异,筛选获得敏感细胞因子TIMP-1、MCP-1、TNF-α、IL-1β、 IFN-γ,并结合文献报道和前期研究,采用Real-Time PCR(实时荧光定量PCR)和ELISA (酶联免疫法)技术,进一步验证分析敏感细胞因子在肺组织中的基因相对表达量和蛋白含量。结果表明,高原组大鼠肺组织中TIMP-1、MCP-1、TNF-α、 IL-1β、IFN-γ的基因相对表达量分别升高2.09、5.23、1.66、1.75、2.57倍,蛋白含量分别升高61 J95%、51.01%、30.77%、12.64%、34.52%(P0.01)。最后,采用布洛芬干预急进高原缺氧大鼠,比较平原组、高原组、布洛芬组和阳性对照药(乙酰唑胺)组大鼠血气、病理变化,测定大鼠肺组织中TIMP-1、MCP-1、TNF-α、L-1β、IFN-γ的基因相对表达量和蛋白含量。结果表明,与高原组比较,布洛芬组缺氧大鼠动脉血PaO2显著性升高4.83%,动脉血乳酸Lac显著性降低51.20%(P0.01),大鼠肺组织损伤明显减轻,高原布洛芬组大鼠肺组织中TIMP-1、MCP-1、TNF-α、IL-1β、IFN-γ的基因相对表达量均显著性下降,蛋白含量分别显著性降低4.75%、24.87%、28.09%、16.86%、14.21%、9.16%(P0.01)。本研究建立了急进高原环境大鼠缺氧模型,采用Antibody Array技术筛选敏感细胞因子,并用ELISA和Real-Time PCR技术进一步验证,利用抗炎药物布洛芬干预,得出以下结论：1、实地缺氧和模拟舱缺氧均会造成大鼠严重肺损伤,与平原组比较,两者血气、生化、病理指标变化趋势基本一致,然而,两个缺氧组之间的血气、生化指标差异明显；2、大鼠急性暴露缺氧环境,激活机体炎症反应,促进细胞因子TIMP-1、 MCP-1、TNF-α、IL-1β、IFN-γ的含量升高,进一步扩大炎症反应,是缺氧刺激造成机体损伤的机制之一；3、布洛芬能够减轻缺氧大鼠肺组织损伤,其保护作用机制可能与布洛芬提高缺氧大鼠Pa02和抑制细胞因子TIMP-1、MCP-1、TNF-α、IL-1β、IFN-γ的表达有关。
[Abstract]:Quickly enter the plateau environment, people often because of hypoxia, nausea, vomiting, weakness and a series of body discomfort, severe cases will lead to acute plateau disease, without early treatment may cause death. However, the pathogenesis of acute high altitude disease is not clear, resulting in drug development is slow, so that the life and health of the people safety suffers the plateau hypoxia environment threat. Therefore, exploring the mechanism of hypoxia injury research plateau, anti hypoxia and protective effect of drugs is very important. This paper will establish a rat model of hypoxia at high altitude field, on the effects of hypoxia on cytokines of rats in vivo, the potential protective effect of anti hypoxia on plateau environment on hypoxia injury in rats and the mechanism study of anti-inflammatory drug ibuprofen. Firstly the hypoxic environment at high altitude and high altitude simulation cabin simulation, establish the rat model of hypoxia, hypoxia group and comparison Blood gas, plain rats, biochemical, pathological differences. The results show that the gas field and high altitude hypoxia in rats at high altitude hypoxia simulation cabin to simulate hypoxia rat biochemical changes, pathological indicators are basically the same, among them, acute hypoxia (simulated hypoxia and hypoxia on the main organ damage field) of rat lung tissue. The results also showed that in blood biochemical research, hypoxia, hypoxia model should be used to study the choice of different models, different results may be larger. The results for the field at high altitude hypoxia model to provide experimental basis for establishing. Secondly, using Antibody Array technology to compare differences in plateau hypoxia group and plain rats in group 34 kinds of cytokines in rats, obtain sensitive cytokine TIMP-1, screening of MCP-1, TNF- alpha, IL-1 beta, IFN- gamma, and combined with the literature and previous studies, using Real-Time (real time PCR Fluorescence quantitative PCR) and ELISA (ELISA) technology, to further verify the analysis of relative expression of gene and protein content of sensitive cytokines in lung tissues. The results showed that the TIMP-1 group in the lung tissue of rats in the MCP-1 plateau, TNF- alpha, IL-1 beta, IFN- gamma gene relative expression quantity increased by 2.09,5.23,1.66,1.75,2.57 times respectively, protein content increased 61 J95%, respectively, 51.01%, 30.77%, 12.64%, 34.52% (P0.01). Finally, the ibuprofen intervention of acute high altitude hypoxia rats, compared with the plain group, plateau group, ibuprofen group and positive control drug (acetazolamide) rats, blood gas, pathological changes, the determination of TIMP-1 in rat lung tissue in MCP-1, TNF- alpha, L-1 beta, relative expression and protein content of IFN- gamma gene. The results showed that compared with the plateau group, arterial blood PaO2 ibuprofen group in hypoxic rats significantly increased 4.83%, arterial blood lactic acid Lac significantly decreased 51.20% (P0.01), lung tissue of rats The damage was significantly reduced, TIMP-1 group lung tissue of rats in plateau ibuprofen MCP-1, TNF- alpha, IL-1 beta, IFN- gamma gene relative expression were significantly decreased, protein content were significantly decreased by 4.75%, 24.87%, 28.09%, 16.86%, 14.21%, 9.16% (P0.01). This study established in rats at high altitude environment hypoxia model, Screening Sensitive cytokines by Antibody Array technology, and further confirmed by ELISA and Real-Time PCR technology, the use of anti-inflammatory drugs ibuprofen intervention, draw the following conclusions: 1. Hypoxia and hypoxia field simulation cabin will cause serious lung injury in rats, compared with the plain group both blood biochemical, pathological change trend index basically the same, however, the blood gas between the two hypoxia group, biochemical index significantly; 2, acute exposure to hypoxia in rats, the activation of the inflammatory reaction, promote cell factor TIMP-1, MCP-1, TNF- alpha, IL-1 beta, gamma IFN- content Increased, further expand the inflammatory reaction is hypoxia caused by one of the mechanisms of body injury; 3, ibuprofen can reduce the lung tissue hypoxia injury in rats, the protective effect and possible mechanism of ibuprofen increased in hypoxic rats Pa02 and inhibition of cytokine TIMP-1, MCP-1, TNF- alpha, IL-1 beta, the expression of IFN- gamma.

【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R594.3

【参考文献】