發(fā)布時(shí)間：2018-05-26 00:49

  本文選題：凝集反應(yīng) + 組織因子��； 參考：《承德醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:目前,病毒感染所引發(fā)的疾病在感染性疾病中所占的比例越來(lái)越大,由于病毒較其他微生物相比具有更強(qiáng)的致病力、感染性和致死率,且尚缺乏有效的預(yù)防、治療手段,引起了某些流感疾病的流行暴發(fā)與傳播,其中以禽流感病毒尤為突出。由于流感病毒表面蛋白的抗原成分頻繁突變及抗原漂移,人類機(jī)體缺乏相應(yīng)的抗體,并且人類可以通過流感疫苗獲得的免疫力微乎其微。盡管存在抗病毒藥物和流感疫苗,世界衛(wèi)生組織估計(jì)僅在發(fā)達(dá)國(guó)家,流感每年可引發(fā)3百萬(wàn)到5百萬(wàn)的危重疾病,導(dǎo)致25萬(wàn)到50萬(wàn)人口死亡,在發(fā)展中國(guó)家可能會(huì)更高。研究發(fā)現(xiàn),流感重癥感染可以造成全身臟器廣泛的病理?yè)p傷及嚴(yán)重的并發(fā)癥,最終導(dǎo)致呼吸衰竭、全身多臟器功能障礙綜合征(multiple organ dysfunction syndrome,MODS)。然而,流感病毒的致病機(jī)制尚不完全明確。研究顯示,流感病毒的致病可能與其引發(fā)的機(jī)體高細(xì)胞因子風(fēng)暴、宿主對(duì)流感病毒的遺傳易感性、病毒血癥有關(guān)。血凝素(Hemagglutinin,HA)是存在于病毒顆粒表面的糖蛋白,在病毒感染過程中起重要作用,是病毒致病力的關(guān)鍵因素。血凝素具有在體內(nèi)外可引發(fā)紅細(xì)胞發(fā)生凝集反應(yīng)的特性,研究表明流感病毒通過凝集特性而向全身播散,我們推測(cè)達(dá)到一定濃度的血凝素引發(fā)的凝集反應(yīng)與病毒的致病相關(guān),可引發(fā)機(jī)體產(chǎn)生一系列的病理生理改變。因此,我們應(yīng)用植物血凝素(phytohemagglutinin,PHA)誘導(dǎo)大鼠體內(nèi)發(fā)生紅細(xì)胞凝集反應(yīng),探索凝集反應(yīng)對(duì)大鼠血漿組織因子(tissue factor,TF)、D-二聚體、IL-6水平及肺部毛細(xì)血管內(nèi)皮細(xì)胞形態(tài)的影響,以期為進(jìn)一步探究流感病毒感染的致病機(jī)制提供新思路。方法:SPF級(jí)健康SD大鼠(雌雄不限,180-200g)共42只,隨機(jī)分為生理鹽水(physiology saline group)對(duì)照組,濃度分別為5mg/ml、10mg/ml、20mg/ml PHA組及相應(yīng)濃度的滅活PHA組,每組6只。通過尾靜脈分別向大鼠體內(nèi)注射生理鹽水、不同濃度的PHA及滅活PHA,劑量為1.5ml/100g體重。給藥4h后,10%水合氯醛腹腔注射麻醉大鼠,下腔靜脈取血,用ELISA法檢測(cè)大鼠血漿中TF、D-二聚體及IL-6的含量。透射電鏡下觀察大鼠肺部毛細(xì)血管內(nèi)皮細(xì)胞的形態(tài)變化。結(jié)果:1.大鼠血漿TF含量:5mg/ml、10mg/ml、20mg/ml植物血凝素組大鼠血漿中的TF含量分別為(123.04±35.96)ng/L、(134.67±48.49)ng/L、(132.78±26.29)ng/L,均顯著高于生理鹽水對(duì)照組(77.28±23.50)ng/L及相應(yīng)濃度的滅活植物血凝素組(74.65±11.44、79.75±10.70、86.56±9.90)ng/L,差異有統(tǒng)計(jì)學(xué)意義(均P0.05)。各濃度植物血凝素組間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.1)。各濃度滅活植物血凝素組大鼠血漿TF含量與生理鹽水對(duì)照組相比,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.1)。2.大鼠血漿D-二聚體含量:5mg/ml、10mg/ml、20mg/ml植物血凝素組大鼠血漿D-二聚體水平(分別為3.12±0.82、2.27±0.74、2.77±0.93)ng/ml,均顯著高于生理鹽水對(duì)照組(0.57±0.35)ng/ml及各相應(yīng)濃度滅活植物血凝素組(0.49±0.25、0.47±0.20、0.48±0.17)ng/ml,均P0.05。各濃度的滅活植物血凝素組大鼠血漿D-二聚體含量與生理鹽水對(duì)照組相比差異無(wú)統(tǒng)計(jì)學(xué)意義(均P0.05)。各濃度植物血凝素組間差異無(wú)顯著性(P0.1)。3.大鼠血漿IL-6的含量:5mg/ml、10mg/ml、20mg/ml植物血凝素組大鼠血漿中的IL-6含量分別為(29.20±9.87)pg/ml、(26.48±4.82)pg/ml、(26.12±6.00)pg/ml,均顯著高于生理鹽水對(duì)照組(17.00±2.72)pg/ml及相應(yīng)濃度的滅活植物血凝素組(17.02±2.54、14.77±1.22、15.07±1.10)pg/ml,差異具有統(tǒng)計(jì)學(xué)意義(均P0.01)。各濃度植物血凝素組間差異無(wú)顯著性(P0.1)。各濃度滅活植物血凝素組與生理鹽水對(duì)照組相比差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.1)。4.大鼠肺部毛細(xì)血管內(nèi)皮細(xì)胞形態(tài):5mg/ml、10mg/ml、20mg/ml濃度植物血凝素組大鼠肺毛細(xì)血管內(nèi)皮細(xì)胞受損,表現(xiàn)為內(nèi)皮細(xì)胞腫脹、溶解,胞質(zhì)疏松,邊界不清,線粒體腫脹,基膜增厚甚至斷裂,含氣小泡減少,且損傷程度隨著植物血凝素濃度的升高而加重。各濃度滅活植物血凝素組與生理鹽水對(duì)照組內(nèi)皮細(xì)胞形態(tài)、結(jié)構(gòu)正常,內(nèi)皮細(xì)胞扁平,邊界清楚,細(xì)胞器結(jié)構(gòu)正常,含氣小泡豐富。結(jié)論:體內(nèi)發(fā)生的紅細(xì)胞凝集反應(yīng)能夠破壞內(nèi)皮細(xì)胞,導(dǎo)致機(jī)體凝血及纖溶系統(tǒng)功能異常,這可能是流感病毒感染體內(nèi)凝血功能及纖溶系統(tǒng)紊亂的機(jī)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.
【學(xué)位授予單位】：承德醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R511.7
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本文編號(hào)：1935298