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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/35071

Title: 探討藻類多醣抗病毒與其免疫調節之作用機制
Study of anti-viral and immunomodulatory effects of polysaccharide extracts from seaweeds on the viral infections
Authors: Ya-Huang Chiu
邱雅凰
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 藻類;多醣;石蓴;鹿藻菜膠;小球藻;日本腦炎病毒
seaweed;polysaccharide;ulva;carrageenan;chlorella;Japanese encephalitis virus
Date: 2012
Issue Date: 2013-10-07T02:50:08Z
Abstract: 藻類多醣萃取物-石蓴多醣 (ulvan)、鹿藻菜膠 (carrageenan)、小球藻多醣萃取物 (Chlorella pyrenoidosa extracts, CPE) 其總醣、硫酸根含量和與抗氧化活性之間並非絕對正相關性。因此推測其活性與硫酸多醣的結構有關。本研究主要是探討石蓴多醣萃取物對日本腦炎病毒感染途徑、其引起發炎的機轉,以及免疫活性調節。實驗結果顯示,石蓴多醣萃取物在抑制日本腦炎病毒的效果相當顯著。由實驗結果推測石蓴主要在抑制日本病毒與細胞接觸、吸收,而非在抑制病毒感染進入細胞後進行複製及病毒顆粒組裝方面。另外在石蓴多醣萃取物抑制日本腦炎感染後誘導 mixed glia cell 產生 NO 之效應上,結果顯示石蓴多醣萃取物確實有抑制日本腦炎感染誘導 mixed glia cell 產生 NO 的現象。在 in vivo 實驗方面,C3H/HeN 品系小鼠在感染日本腦炎病毒後第五天出現背部拱起、後肢癱瘓麻痺的情形,並於感染後第七天死亡,而石蓴多醣萃取物餵食的組別,可以延緩小鼠發病及死亡的情形。在評估鹿藻菜膠是否可以有效對抗腸病毒所引起的感染部分。鹿藻菜膠可以顯著並有效的抑制病毒斑的形成。在病毒吸附階段前或病毒吸附階段中,添加鹿藻菜膠可有效抑制病毒 RNA的複製。鹿藻菜膠的處理可間接抑制病毒感染所引起的細胞凋亡。在病毒吸附能力試驗,顯示出鹿藻菜膠可以與病毒顆粒產生吸附作用。電子顯微鏡結果顯示鹿藻菜膠可以吸附至病毒顆粒外圍,藉以阻擋病毒顆粒與細胞膜上的接受器結合。在評估小球藻多醣萃取物及鹿角菜膠對於第二型登革病毒感染是否具抑制效果。細胞實驗方面,小球藻多醣萃取物及鹿角菜膠可干擾病毒感染之早期階段。小球藻多醣萃取物及鹿角菜膠直接與病毒顆粒作用。第二型登革病毒之 C3H/HeN 乳鼠感染模式中,八日齡小鼠經顱內注射 100 PFU/mouse 之病毒感染劑量,死亡率為 100%,並有部分感染小鼠出現腸胃道出血、血漿滲漏的登革症狀。小球藻及鹿角菜膠於登革病毒攻毒前混合,可提高感染小鼠之存活率。在利用細胞實驗及小鼠動物模式探討藻類多醣萃取物對於 A 型流感病毒感染抑制效果。細胞實驗依病毒感染吸附階段、吸附前、後進行藻類多醣萃取物-石蓴多醣、鹿藻菜膠、小球藻多醣萃取物處理,結果顯示 藻類多醣萃取物在吸附階段有較佳的保護效果。 BALB/c 小鼠感染模式中,6 週齡大幼鼠感染致死劑量 (100 PFU/mouse) 其死亡率 100%,並在病毒感染前藻類多醣萃取物直接與病毒顆粒吸附處理再進行感染,能有效降低小鼠死亡率。並可有效減少病毒感染小鼠後發炎相關之細胞激素 (cytokines) 及趨化激素 (chemokines) 的生成量。綜合上述的研究成果,藻類多醣萃取物在細胞試驗及動物試驗中皆具有抑制病毒感染的能力。是具有發展成抗病毒保健食品之潛力。
The seaweed polysaccharide extracts of ulvan, carrageenan, (Chlorella pyrenoidosa extracts, CPE). The total sugars, sulfate contents and molecular weights with no direct correlations. The diverse activities may depend on given structures in the sulfated polysaccharides. In this study, we identified that the polysaccharide extracts from Ulva can inhibit JEV infection in Vero cells. Mechanistic studies further revealed that the Ulva polysaccharide extracts can block virus adsorption and thus make the virus unable to enter cells. The Ulva polysaccharide extracts also effectively decrease the production of proinflammatory cytokines in the JEV-infected primary mixed glia cells. In animal study, the JEV-infected C3H/HeN mice appeared neurobehavioral abnormalities at the 5th day and died at the 7th day post-infection. However, the JEV infected mice pretreated with the Ulva polysaccharide extracts can delay the onset of hind limb paralysis and thereby prevent the mice from death. In this study, we reported carrageenan also has a strong and effective anti-EV 71 activity able to reduce plaque formation, prevent viral replication before or during viral adsorption, as well as inhibit EV 71-induced apoptosis. In virus binding assay, carrageenan was shown able to bind EV 71 firmly, forming carrageenan-viruses complexes, whereby the virus-receptor interaction is likely disrupted. In this study, the antiviral activity of hot water extracts from microalgae (Chlorella pyrenoidosa extracts, CPE) and carrageenan on dengue virus type II (DV2) was evaluated. In vitro antiviral studies, the CPE and carrageenan could interference the early step of dengue virus infection. And CPE and carrageenan direct react with DV2 virus particles was better than other treatments. In the DV2 infected C3H/HeN mice model, all of eighth-day-old mice would die within 6 - 8 days after intracranial inoculation of DV2 with 100 PFU/mouse, some of them developed the typical dengue syndrome such as plasma leakage and gastrointestinal hemorrhagic. In vivo antiviral studies, eighth-day-old mice inoculate with mixture of DV2 and CPE or carrageenan could elevate the survival rate. In this study investigated that seaweed polysaccharide extracts of ulvan, carrageenan, CPE for inhibiting influenza A virus infection on in vitro and in vivo model. In vitro, the batter inhibition effect of virus adsorption on influenza virus infection is during adsorption stage. In BALB/c mice animal model, the 6 weeks-old mice infecte with lethal doses influenza virus (100 PFU/mouse) and the mortality rate is 100%. The mice treatment seaweed polysaccharide extracts adsorption with virus before infection, the mortality rate of these mice are effectively reduced. The seaweed polysaccharide extracts also effectively decrease the production of proinflammatory cytokines and chemokines in the H1N1-infected mice. In summary, the seaweed polysaccharide extracts show a protective effect against virus infection on both in cell and animal experiment. Seaweed polysaccharides extract may be an ideal candidate worth while to develop into anti-viral agents.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0D96320002
http://ntour.ntou.edu.tw/handle/987654321/35071
Appears in Collections:[食品科學系] 博碩士論文

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