凝集反应对大鼠血浆组织因子、D-二聚体、IL-6水平及内皮细胞的影响

发布时间：2018-05-26 00:49

  本文选题：凝集反应 + 组织因子　； 参考：《承德医学院》2017年硕士论文

【摘要】：目的:目前,病毒感染所引发的疾病在感染性疾病中所占的比例越来越大,由于病毒较其他微生物相比具有更强的致病力、感染性和致死率,且尚缺乏有效的预防、治疗手段,引起了某些流感疾病的流行暴发与传播,其中以禽流感病毒尤为突出。由于流感病毒表面蛋白的抗原成分频繁突变及抗原漂移,人类机体缺乏相应的抗体,并且人类可以通过流感疫苗获得的免疫力微乎其微。尽管存在抗病毒药物和流感疫苗,世界卫生组织估计仅在发达国家,流感每年可引发3百万到5百万的危重疾病,导致25万到50万人口死亡,在发展中国家可能会更高。研究发现,流感重症感染可以造成全身脏器广泛的病理损伤及严重的并发症,最终导致呼吸衰竭、全身多脏器功能障碍综合征(multiple organ dysfunction syndrome,MODS)。然而,流感病毒的致病机制尚不完全明确。研究显示,流感病毒的致病可能与其引发的机体高细胞因子风暴、宿主对流感病毒的遗传易感性、病毒血症有关。血凝素(Hemagglutinin,HA)是存在于病毒颗粒表面的糖蛋白,在病毒感染过程中起重要作用,是病毒致病力的关键因素。血凝素具有在体内外可引发红细胞发生凝集反应的特性,研究表明流感病毒通过凝集特性而向全身播散,我们推测达到一定浓度的血凝素引发的凝集反应与病毒的致病相关,可引发机体产生一系列的病理生理改变。因此,我们应用植物血凝素(phytohemagglutinin,PHA)诱导大鼠体内发生红细胞凝集反应,探索凝集反应对大鼠血浆组织因子(tissue factor,TF)、D-二聚体、IL-6水平及肺部毛细血管内皮细胞形态的影响,以期为进一步探究流感病毒感染的致病机制提供新思路。方法:SPF级健康SD大鼠(雌雄不限,180-200g)共42只,随机分为生理盐水(physiology saline group)对照组,浓度分别为5mg/ml、10mg/ml、20mg/ml PHA组及相应浓度的灭活PHA组,每组6只。通过尾静脉分别向大鼠体内注射生理盐水、不同浓度的PHA及灭活PHA,剂量为1.5ml/100g体重。给药4h后,10%水合氯醛腹腔注射麻醉大鼠,下腔静脉取血,用ELISA法检测大鼠血浆中TF、D-二聚体及IL-6的含量。透射电镜下观察大鼠肺部毛细血管内皮细胞的形态变化。结果:1.大鼠血浆TF含量:5mg/ml、10mg/ml、20mg/ml植物血凝素组大鼠血浆中的TF含量分别为(123.04±35.96)ng/L、(134.67±48.49)ng/L、(132.78±26.29)ng/L,均显著高于生理盐水对照组(77.28±23.50)ng/L及相应浓度的灭活植物血凝素组(74.65±11.44、79.75±10.70、86.56±9.90)ng/L,差异有统计学意义(均P0.05)。各浓度植物血凝素组间比较差异无统计学意义(P0.1)。各浓度灭活植物血凝素组大鼠血浆TF含量与生理盐水对照组相比,差异无统计学意义(P0.1)。2.大鼠血浆D-二聚体含量:5mg/ml、10mg/ml、20mg/ml植物血凝素组大鼠血浆D-二聚体水平(分别为3.12±0.82、2.27±0.74、2.77±0.93)ng/ml,均显著高于生理盐水对照组(0.57±0.35)ng/ml及各相应浓度灭活植物血凝素组(0.49±0.25、0.47±0.20、0.48±0.17)ng/ml,均P0.05。各浓度的灭活植物血凝素组大鼠血浆D-二聚体含量与生理盐水对照组相比差异无统计学意义(均P0.05)。各浓度植物血凝素组间差异无显著性(P0.1)。3.大鼠血浆IL-6的含量:5mg/ml、10mg/ml、20mg/ml植物血凝素组大鼠血浆中的IL-6含量分别为(29.20±9.87)pg/ml、(26.48±4.82)pg/ml、(26.12±6.00)pg/ml,均显著高于生理盐水对照组(17.00±2.72)pg/ml及相应浓度的灭活植物血凝素组(17.02±2.54、14.77±1.22、15.07±1.10)pg/ml,差异具有统计学意义(均P0.01)。各浓度植物血凝素组间差异无显著性(P0.1)。各浓度灭活植物血凝素组与生理盐水对照组相比差异无统计学意义(P0.1)。4.大鼠肺部毛细血管内皮细胞形态:5mg/ml、10mg/ml、20mg/ml浓度植物血凝素组大鼠肺毛细血管内皮细胞受损,表现为内皮细胞肿胀、溶解,胞质疏松,边界不清,线粒体肿胀,基膜增厚甚至断裂,含气小泡减少,且损伤程度随着植物血凝素浓度的升高而加重。各浓度灭活植物血凝素组与生理盐水对照组内皮细胞形态、结构正常,内皮细胞扁平,边界清楚,细胞器结构正常,含气小泡丰富。结论:体内发生的红细胞凝集反应能够破坏内皮细胞,导致机体凝血及纤溶系统功能异常,这可能是流感病毒感染体内凝血功能及纤溶系统紊乱的机制之一。
[Abstract]:Objective: at present, the proportion of diseases caused by virus infection is more and more serious in infectious diseases. Because the virus has stronger pathogenicity, infectivity and lethality compared with other microorganisms, the virus is still lack of effective prevention, and the treatment means cause epidemic outbreak and spread of some influenza diseases, especially the avian influenza virus. Because of the frequent mutation of the antigen components of the surface protein of the influenza virus and the drift of the antigen, the human body lacks the corresponding antibody, and the immunity obtained by the influenza vaccine is very small. Despite the presence of antiviral drugs and influenza vaccines, the WHO estimates that in developed countries only 3 million to 5 of the flu can be triggered a year. Millions of critical diseases, which cause 250 thousand to 500 thousand people to die, may be higher in developing countries. Studies have found that severe influenza infection can cause extensive pathological damage and severe complications in the whole body, resulting in respiratory failure, multiple organ dysfunction syndrome, MODS. The pathogenic mechanism of influenza virus is not yet completely clear. The study shows that the pathogenesis of influenza virus may be related to the high cytokine storm caused by the virus, the host's genetic susceptibility to influenza virus and the viremia. Hemagglutinin (HA) is a glycoprotein that exists in the surface of the virus particles, and plays an important role in the virus infection process. It is a key factor in the virulence of the virus. Hemagglutinin has the characteristics that can cause agglutination of erythrocytes in the body and the body. The study shows that the influenza virus spread through the agglutination. We speculate that the agglutination induced by a certain concentration of hemagglutinin is associated with the pathogenicity of the virus, which can cause a series of pathogeny. Therefore, we used phytohemagglutinin (PHA) to induce erythrocyte agglutination in rats and explore the effect of agglutination on the plasma tissue factor (tissue factor, TF), D- two polymer, IL-6 level and pulmonary capillary endothelial cell morphology in order to further explore the cause of influenza virus infection. Methods: a total of 42 healthy SD rats (male and female, 180-200g) were randomly divided into normal saline (physiology saline group) control group, and the concentration was 5mg/ml, 10mg/ml, 20mg/ml PHA group and the corresponding concentration of inactivated PHA group, 6 rats in each group. The saline was injected into the rat body through the tail vein, and the concentration was different. PHA and inactivated PHA, dose 1.5ml/100g weight. After administration of 4h, 10% chloral hydrate was intraperitoneally injected with anesthetized rats, and blood was taken in the inferior vena cava. The content of TF, D- two polymer and IL-6 in rat plasma was detected by ELISA. The morphological changes of pulmonary capillary endothelial cells in rats were observed under transmission electron microscope. Results: TF content in plasma of 1. rats: 5mg/ml, 10mg/ml, 20mg/ The content of TF in the plasma of ML hemagglutinin group was (123.04 + 35.96) ng/L, (134.67 + 48.49) ng/L and (132.78 + 26.29) ng/L, which were significantly higher than that in the saline control group (77.28 + 23.50) ng/L and the corresponding concentration of inactivated plant hemagglutinin group (74.65 + 11.44,79.75 + 10.70,86.56 + 9.90) ng/L, the difference was statistically significant (P0.05). There was no significant difference between the plant hemagglutinin groups (P0.1). The plasma TF content of the rats in the inactivated plant hemagglutinin group was not significantly different from that of the normal saline control group (P0.1), the plasma D- two polymer content in.2. rats: 5mg/ml, 10mg/ml, 20mg/ml plant hemagglutinin group rats plasma D- two polymer level (3.12 + 0.82,2.27 respectively) The 0.74,2.77 + 0.93) ng/ml was significantly higher than that of the normal saline control group (0.57 + 0.35) ng/ml and the corresponding concentration inactivated plant hemagglutinin group (0.49 + 0.25,0.47 + 0.20,0.48 + 0.17) ng/ml, and the plasma D- two polymer content of the inactivated plant hemagglutinin group was not significantly different from that of the control group (all P0.05). There was no significant difference between the concentration of hemagglutinin groups in each concentration (P0.1).3. rat plasma IL-6 content: the IL-6 content in the plasma of 5mg/ml, 10mg/ml, 20mg/ml plant hemagglutinin group was (29.20 + 9.87) pg/ml, (26.48 + 4.82) pg/ml and (26.12 + 6) pg/ml, which were significantly higher than that in the saline control group (17 + 2.72) pg/ml and the inactivation of the corresponding concentration. The difference of hemagglutinin group (17.02 + 2.54,14.77 + 1.22,15.07 + 1.10) pg/ml was statistically significant (P0.01). There was no significant difference between the concentration of plant hemagglutinin groups (P0.1). There was no significant difference between the inactivated plant hemagglutinin group and the normal saline control group (P0.1) the pulmonary capillary endothelial cell morphology of the lung of.4. rats: 5mg/ml, 10 Mg/ml, 20mg/ml concentration plant hemagglutinin group rats lung capillary endothelial cells damage, manifested as endothelial cells swelling, dissolved, loose cytoplasm, indistinct boundaries, mitochondria swelling, thickening even fracture of the basement membrane, and decreasing gas vesicles, and the degree of injury aggravated with the increase of plant hemagglutinin concentration. The endothelial cells of the control group were normal, the endothelial cells were flat, the boundary was clear, the organelle structure was normal, and the gas containing vesicles were abundant. Conclusion: the erythrocyte agglutination reaction in the body can destroy the endothelial cells and lead to the abnormal function of the blood coagulation and fibrinolysis system, which may be the coagulation function and fibrinolytic system of influenza virus infection in the body. One of the mechanisms of disorder.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R511.7
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本文编号：1935298