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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/48858

Title: λ-鹿角菜膠及其硫酸根衍生物對於腸病毒71型感染之保護效果
The protective effects of λ-carrageenan and its sulfated derivatives on enterovirus 71 infection
Authors: Tsai, Chia-Yun
Contributors: NTOU:Department of Food Science
Keywords: 腸病毒 71 型;λ-鹿角菜膠;硫酸根衍生物
Enterovirus 71;λ-carrageenan;sulfated derivatives
Date: 2014
Issue Date: 2018-08-22T06:38:42Z
Abstract: 腸病毒71型 (EV71) 分類學上為屬於小核糖核酸病毒科,腸病毒屬,臨床症狀除產生手足口症 (hand-foot-mouth disease) 外,易引發中樞神經系統疾病,引起孩童死亡,目前仍無有效藥物或疫苗,因此目前許多研究積極從日常生活中找尋有效對抗腸病毒感染之活性物質。海藻硫酸多醣已被證實能有效降低許多病毒感染能力,研究也發現改變多醣上硫酸根含量會影響其生理活性。因此本論文利用實驗室建立之 carrageenan (CGN) 降低 EV71 感染模式,進一步針對不同型態 CGN 對於 EV71 感染細胞之抑制率,並篩選出效果最佳 CGN 進行後續實驗。利用ICGN、LCGN 及 KCGN 以其 IC50 進行樣品篩選,並以病毒斑測試再次驗證,篩選出 LCGN 抗 EV71 效果最佳。因此選定 LCGN 作為後續實驗樣品,並製備 LCGN 硫酸根衍生物 LCGN-S 與 LCGN-DS,其硫酸基含量分別為 24%、 32% 與 18%。在細胞實驗中發現在1,000 g/ml 下 LCGN 及LCGN-S、LCGN-DS 皆具有顯著降低病毒抑制率,並以 LCGN 與 LCGN-DS 效果最佳。進一步利用免疫螢光試驗測定在共同培養條件下胞內病毒抗原 VP1 表現量,其趨勢也相同。病毒吸附能力試驗中也顯示其吸附能力為:LCGN > LCGN-DS > LCGN-S。此外 LCGN 處理後降低感染後細胞內 ROS、caspase 活性,也降低病毒感染所導致細胞凋亡現象。於我們目前研究顯示,硫酸多醣其抗病毒效果可能不只取決於硫酸根含量,也與其分子量大小及硫酸根所在醣基位置有關。LCGN 可能藉由阻擋 EV71 感染細胞進而降低細胞凋亡相關因子表現,而非具有治療效果。
Enterovirus 71 (EV71) belongs to the Enterovirus genus of picornaviridae. EV71 infections are usually associated with hand- foot- mouth disease in young children and may also cause severe neurological complications with high mortality rates. There is still no effective drugs or vaccines against virus infection. Hence, there is an urgent need to develop the specific anti-enterovirus 71 agents from local resources. Sulfated polysaccharides have been proven to reduce the ability of viral infections, and the biological activities changed by modification of the sulfated group on the polysaccharides has also been reported. In this study, we followed the model of anti-EV71 effects of -carrageenan on vero cells from our laboratory. First, we used ICGN, LCGN, KCGN for screening and selecting the most effective LCGN as the sample of our study by IC50 and plaque assay. The sulfate content of LCGN and its derivatives LCGN-S and LCGN-DS were 24%, 32%, and 18%. In virus binding assay, the binding abilities were: LCGN > LCGN-DS > LCGN-S when treated with 1,000 g/ml. LCGN also showed reduce the viral induced ROS expression, caspase activities and apoptosis. Consequently, given our present findings, we propose that the anti-EV71 acivity of CGN not only depends on the sulfate content, but also the molecule weights and the position of ulfate on the polysaccharides. LCGN reduced the viral induced ROS expression, caspase activities and apoptosis by binding with EV71, avoiding EV71 infections to the host cells.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010132044.id
Appears in Collections:[食品科學系] 博碩士論文

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