本文關(guān)鍵詞：乙肝、肝硬化、肝癌患者與流感病毒易感性關(guān)聯(lián)的研究　出處：《西北大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 流感病毒 唾液 血凝素(HA) 乙肝 肝硬化 肝癌

【摘要】：研究背景及意義：研究表明,流感病毒感染宿主的過(guò)程中,主要是通過(guò)血凝素(HA)與宿主細(xì)胞表面唾液酸糖鏈?zhǔn)荏w進(jìn)行特異性的結(jié)合。人流感病毒HA主要通過(guò)識(shí)別并結(jié)合SA α2-6Gal的糖鏈?zhǔn)荏w,而禽流感病毒HA主要識(shí)別并結(jié)合SA α2-3Gal的糖鏈?zhǔn)荏w。已有研究報(bào)道,在人呼吸道上皮細(xì)胞表面含有著SA α2-6Gal糖鏈末端結(jié)構(gòu)和少量的SAα2-3Gal糖鏈末端結(jié)構(gòu)。 唾液中含有豐富的糖蛋白,它們具有抗菌、抗病毒以及潤(rùn)滑口腔等作用。近年來(lái)有研究顯示,疾病的發(fā)生涉及到了唾液糖蛋白糖鏈結(jié)構(gòu)的改變。唾液是防止呼吸道病毒進(jìn)入人體的首要防線。其中,唾液中的糖蛋白糖鏈末端結(jié)構(gòu)在抵御病毒中起著決定性的作用,其可以與病毒特異性的結(jié)合而達(dá)到防御目的。 本實(shí)驗(yàn)室研究發(fā)現(xiàn),健康志愿者中,老年人與兒童、年輕人相比較,老年人具有更強(qiáng)抵抗流感的能力。其主要分子機(jī)制之一,是由于老年人唾液與兒童、年輕人唾液相比較,具有更多的SAα2-3/SA α2-6Gal糖鏈末端結(jié)構(gòu)。這些糖鏈末端結(jié)構(gòu)可以結(jié)合更多的流感病毒血凝素(HA)。但流感爆發(fā)時(shí),臨床上發(fā)現(xiàn)患者多為患有慢性疾病老年人?其原因是慢性疾病破壞了老年人的免疫系統(tǒng),使其抵抗力降低。除此原因外,是否有其他生理或病理的因素導(dǎo)致老年人更易感流感病毒?乙肝、肝硬化和肝癌是常見(jiàn)的幾種慢性疾病,嚴(yán)重威脅著人類的健康,患有這幾種慢性疾病的人群是否更易感流感病毒?感染的特性又是什么?這是本課題的立題依據(jù),也是公共衛(wèi)生領(lǐng)域亟需解決的一些問(wèn)題。 方法：首先,收集乙肝、肝硬化、肝癌患者和健康志愿者唾液樣本,然后通過(guò)凝集素芯片技術(shù),對(duì)識(shí)別SAα2-6Gal糖鏈末端結(jié)構(gòu)的凝集素SNA和識(shí)別SAα2-3Gal糖鏈末端結(jié)構(gòu)MAL-II所對(duì)應(yīng)的的熒光信號(hào)值進(jìn)行分析、比較,最終得到乙肝、肝硬化、肝癌患者和健康志愿者唾液中SAα2-6Gal和SAα2-3Gal糖蛋白糖鏈末端結(jié)構(gòu)表達(dá)水平的差異性。隨后,利用印記技術(shù),將凝集素SNA、MAL-Ⅱ和雞源H5N1流感病毒、鴨源H5N1流感病毒、雞源H9N2流感病毒以及甲型H1N1流感病毒裂解疫苗分別與乙肝、肝硬化、肝癌患者和健康志愿者的唾液樣本進(jìn)行印記實(shí)驗(yàn)。根據(jù)實(shí)驗(yàn)結(jié)果,對(duì)乙肝、肝硬化、肝癌患者與流感病毒易感性關(guān)聯(lián)進(jìn)行分析討論。 結(jié)果：1.應(yīng)用凝集素芯片技術(shù),對(duì)乙肝、肝硬化、肝癌患者和健康志愿者唾液進(jìn)行檢測(cè),分析其中唾液酸化糖蛋白糖鏈結(jié)構(gòu)表達(dá)水平變化的情況。通過(guò)分析乙肝、肝硬化和肝癌患者樣本的熒光信號(hào)值,并將結(jié)果與健康志愿者唾液樣本熒光信號(hào)值進(jìn)行比對(duì),從而得出比值Ratio值。結(jié)果顯示,在乙肝、肝硬化和肝癌患者的唾液樣本中,凝集素MAL-Ⅱ?qū)?yīng)的Ratio值分別為0.423、0.437和0.479。因此,MAL-II所識(shí)別的SAα2-3Gal糖鏈末端結(jié)構(gòu)在乙肝、肝硬化和肝癌患者唾液中的表達(dá)水平明顯下降。而在乙肝、肝硬化和肝癌患者唾液中,凝集素SNA對(duì)應(yīng)的Ratio值分別為2.294、1.409和1.336,因此,其所識(shí)別的SAα2-6Gal糖鏈末端結(jié)構(gòu)在乙肝、肝硬化和肝癌患者唾液中的表達(dá)水平明顯上調(diào)。 2.應(yīng)用印記技術(shù),將雞源H5N1流感病毒、鴨源H5N1流感病毒和雞源H9N2流感病毒,分別與乙肝、肝硬化、肝癌患者和健康志愿者的唾液樣本相結(jié)合。通過(guò)三種禽流感病毒與唾液糖蛋白結(jié)合的印記條帶,可以明顯觀察到：鴨源H5N1流感病毒、雞源H5N1流感病毒和雞源H9N2流感病毒與乙肝、肝硬化以及肝癌患者的唾液糖蛋白的結(jié)合能力明顯要弱于與健康志愿者的結(jié)合能力。并將其結(jié)果與凝集素MAL-Ⅱ與乙肝、肝硬化、肝癌患者和健康志愿者的唾液糖蛋白結(jié)合的印記條帶作對(duì)比,發(fā)現(xiàn)三種禽流感病毒的印跡條帶與其保持一致。說(shuō)明在乙肝、肝硬化以及肝癌患者唾液糖蛋白中,禽流感病毒HA特異性識(shí)別、結(jié)合的的SAα2-3Gal糖鏈末端結(jié)構(gòu)表達(dá)水平較低。而凝集素SNA與乙肝、肝硬化以及肝癌患者唾液蛋白的結(jié)合能力強(qiáng)于與健康志愿者的結(jié)合能力,說(shuō)明在乙肝、肝硬化以及肝癌患者唾液蛋白中,人流感病毒表面HA特異識(shí)別、結(jié)合的SAα2-6Gal糖鏈末端結(jié)構(gòu)表達(dá)水平較高。同時(shí),甲型H1N1流感病毒裂解疫苗(甲型H1N1流感病毒株(2009),包含有禽流感、豬流感和人流感三種流感病毒的核糖核酸基因片斷,同時(shí)識(shí)別禽流感病毒糖鏈?zhǔn)荏w和人流感病毒糖鏈?zhǔn)荏w)與四種唾液蛋白樣本的結(jié)合能力無(wú)顯著性差異。因此結(jié)果表明,乙肝、肝硬化和肝癌患者屬于禽流感病毒易感人群,易感禽流感；對(duì)人流感病毒有正常的抵抗力。
[Abstract]:The research background and significance: the study shows that the process of influenza virus infection in the host, mainly through the hemagglutinin (HA) combined with specific host cell surface sialic acid sugar chain receptor. The human influenza virus HA by recognizing and binding to the sugar chain SA receptor alpha 2-6Gal, and avian influenza virus HA mainly recognize and bind sugar chain SA receptor alpha 2-3Gal. It has been reported that on the surface of human airway epithelial cells containing the SA alpha 2-6Gal sugar chain end structure and a small amount of SA alpha 2-3Gal sugar chain structure at the end.
The saliva contains rich glycoprotein, antibacterial, antiviral and lubricating oral effects. In recent years studies have shown that the occurrence of diseases related to salivary glycoprotein glycan structure change. Saliva is to prevent respiratory virus into the first line of the body. The glycoprotein glycan structure at the end of saliva in the decisive role in resisting the virus, the virus can be combined with specific to defensive purposes.
This laboratory study found that in healthy volunteers, the elderly and children, young people compared to older people with stronger ability to resist the flu. One of the major molecular mechanism is due to the elderly and young children saliva, saliva compared with more SA alpha 2-3/SA alpha 2-6Gal sugar chain structure at the end of the sugar chain. Terminal structure can be combined with the influenza virus hemagglutinin (HA). But more of an influenza outbreak, clinically found patients are suffering from chronic diseases in elderly people? The reason is that chronic disease destroys the immune system of the elderly, so that the resistance decreased. In addition to this reason, if there are other factors of physiological or pathological in the elderly people are more susceptible to the flu virus? Hepatitis B, cirrhosis and HCC are several common chronic diseases, serious threat to human health, it suffers from several chronic diseases in populations are more susceptible to influenza virus infection characteristics? What is it? This is the basis of this topic, and it is also an urgent problem to be solved in the field of public health.
Method: first, collect hepatitis B, liver cirrhosis, liver cancer patients and healthy volunteers saliva samples, followed by lectin microarray technology, to identify the SA alpha 2-6Gal sugar chain structure at the end of the lectin SNA and identification of SA alpha 2-3Gal sugar chain structure at the end of MAL-II corresponding to the value of the fluorescence signal for analysis, comparison, finally get hepatitis B, cirrhosis of the liver. The level of expression of SA and SA alpha 2-6Gal alpha 2-3Gal glycoprotein glycan structure at the end of HCC patients and healthy volunteers in the saliva. Then, using imprint technology, the lectin SNA, MAL- II and H5N1 avian influenza virus H5N1 influenza virus, duck, chicken H9N2 influenza virus and influenza a H1N1 and Influenza Split Vaccine were hepatitis B cirrhosis HCC patients and healthy volunteers, saliva samples mark experiment. According to the experimental results of hepatitis B, liver cirrhosis, liver cancer and influenza virus susceptibility association analysis Discuss.
Results: 1. cirrhosis by lectin microarray technology for hepatitis B, liver cancer patients and healthy volunteers saliva were detected, analyzed the expression of sialylated glycoprotein glycan structure. Through the analysis of hepatitis B, cirrhosis and hepatocellular carcinoma samples of fluorescence signal values, and the results of healthy volunteers with a saliva sample fluorescence signal value comparison so that the ratio of Ratio value. The results showed that in hepatitis B, cirrhosis and liver cancer with the saliva samples, the corresponding Ratio II lectin MAL- values were 0.423,0.437 and 0.479. so SA alpha 2-3Gal sugar chain structure at the end of MAL-II identified in the saliva of patients with hepatitis B, the expression level of liver cirrhosis and hepatocellular carcinoma in hepatitis B decreased significantly. Saliva, liver cirrhosis and liver cancer patients, lectin SNA Ratio values were 2.294,1.409 and 1.336, therefore, at the end of the SA alpha 2-6Gal sugar chain recognition The expression of the structure in the saliva of the patients with hepatitis B, liver cirrhosis and liver cancer was obviously up-regulated.
2. application of imprinting, the chicken origin H5N1 influenza virus, influenza virus and duck H5N1 avian H9N2 influenza virus, and hepatitis B cirrhosis, respectively, HCC patients and healthy volunteers saliva samples combined. Through the combination of a mark of three avian influenza virus and salivary glycoprotein bands can be observed clearly: duck origin influenza H5N1 virus, influenza virus and hepatitis B avian H5N1 influenza virus and chicken H9N2, cirrhosis and hepatocellular carcinoma patients with salivary glycoprotein binding ability was weaker than the binding ability and healthy volunteers. And the results of the lectin MAL- II and hepatitis B, liver cirrhosis, liver cancer patients and healthy volunteers with salivary glycoprotein bands as mark contrast, found three bird flu virus was consistent with the band. In hepatitis B, cirrhosis and HCC saliva glycoprotein, HA avian influenza virus specific identification, node SA alpha 2-3Gal sugar chain structure at the end of the low expression of lectins SNA and hepatitis B, cirrhosis and liver cancer patients with salivary protein ability in healthy volunteers with the binding capability in hepatitis B, cirrhosis and HCC saliva protein, people sense virus surface HA specific recognition, SA alpha 2-6Gal sugar end of the chain structure with higher expression level. At the same time, Influenza Split Vaccine (A / H1N1 influenza a H1N1 influenza virus strain (2009), including avian influenza, DNA fragments of three influenza virus swine and human influenza, and identification of avian influenza virus and human influenza virus receptor carbohydrate carbohydrate receptor) had no significant difference and combining ability of four kinds of salivary protein samples. The results showed that hepatitis B, cirrhosis and liver cancer patients belonging to avian influenza virus susceptible, susceptible to avian influenza; a normal flu virus on abortion Resistance.

【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R511.7;R735.7;R512.62

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