褐藻糖胶Fucoidan 对肿瘤细胞的抑制作用.ppt

行政院農業委員會水產試驗所 黃培安,褐藻醣膠(Fucoidan) 對於腫瘤細胞之抑制作用,What is Fucoidan？！,The polysaccharide was named as “fucoidin” when it was first isolated from marine brown algae by Kylin in 1913. but some also called it fucan, fucosan or sulfated fucan. Now it is named as “fucoidan” according to IUPAC rules. Fucoidans are sulfated polysaccharides extracted from marine brown seaweed. (Matou et al., 2002) Fucoidan is major composing of fucose, uronic acid, galactose, xylose and sulphated fucose. (Marais and Joseleau, 2001),The chemical compositions of most fucoidans are complex. The structures of fucoidans from different brown algae vary from species to species. (Li et al., 2008) Fucoidan contains a substantial percentage of L-fucose and sulfate ester groups, and the structural characteristics of fucoidan are very similar to those of heparin. (Yang et al., 2006) Compared with other sulfated polysaccharides, fucoidans are widely available from various kinds of cheap sources, so more and more fucoidans have been investigated in recent years to develop the drugs or functional foods. (Li et al., 2008),Where is Fucoidan from？！,What we have in Taiwan？！,Method 3.,Method 2.,Method1.,Active,1.誘導癌細胞凋亡（Apoptosis） 2.活化免疫作用 3.抑制血管新生作用,ANTI-TUMOR?!,（A）,（B）,圖、Fucoidan處理人類肝癌細胞之細胞型態變 化，箭頭處為細胞型態不完整。（A）控制組、（B） Fucoidan。,Fucoidan (mg/ml),1. 誘導癌細胞凋亡（Apoptosis）,圖、Fucoidan對於人類肝癌細胞株 Hep G2 細胞週期之影響。 (A) medium (B) 1 mg/ml (C) 2.5 mg/ml (D) 5 mg/ml。,4.8%,39.5%,54.2%,56.8%,在細胞週期中： G1時期就是細胞開始休息。 S 時期，DNA開始準備合成。 M時期則準備分裂。 將細胞週期停留在Sub-G1時期，則細胞不會繼續分裂，而沒法增殖，終至凋亡。,凋亡晚期,凋亡早期,圖、磷脂絲氨酸(PS)外翻(Annexin V-FITC/PI 雙染法)分析Fucoidan對於人類肝癌細胞株 Hep G2 誘導之細胞凋亡。 (A) medium (B) 1 mg/ml (C) 2.5 mg/ml (D) 5 mg/ml。,圖、Fucoidan對於人類肝癌細胞株 Hep G2對於人類肝癌細胞株 Hep G2 caspases 活性之影響。 (A) medium (B) 1 mg/ml (C) 2.5 mg/ml (D) 5 mg/ml。,Caspases引發一連串蛋白質酶解作用，傳遞死亡訊息給下游酵素，進一步造成DNA的片段化。,圖、Fucoidan對於人類肝癌細胞株 Hep G2 DNA ladder 之影響,DNA已被切成小片段，因此在DNA凝膠分析時，可以觀察到DNA階層的現象。,2. 活化免疫作用,圖、Fucoidan對於免疫細胞HB4C5之影響,圖、Fucoidan對於免疫細胞HB4C5分泌IgM抗體之影響,圖、Fucoidan對於吞噬細胞J774.1之影響,圖、Fucoidan對於吞噬細胞J774.1吞噬能力之影響,圖、Fucoidan對於小鼠自然殺手細胞之影響,圖、Fucoidan對於人類肝癌細胞株 Hep G2 血管新生相關基因 VEGF 、FLK-1、Tie-2 receptor 與 Angiopoietin-1 基因表現之影響。,3. 抑制血管新生作用,Inflammatory V.S Tumor formation,NF-B,Cell Apoptosis,NF-B 是發炎反應過程中極為重要的分子，當細胞接受到來自細胞外的訊息刺激時，會呈現活化狀態。 活化之後的NF-B 進入細胞核中並引發特定基因轉錄及表現，產生後續發炎反應、調控細胞凋亡及進一步導致腫瘤之發生。,TNF-,IL-1 (g/ml),IL-6 (g/ml),TNF- (g/ml),Fig. Effect of Fucoidan on the secretion of IL-1, IL-6 and TNF- by LPS-stimulated RAW264.7 cells (n=3).,1g/ml LPS,IL-1/-actin fold,iNOS/-actin fold,COX-2/-actin fold,Fig. Effect of Fucoidan on the mRNA expression of IL- (A), iNOS (B) and COX-2 (C) by LPS-stimulated RAW264.7 cells (n=3).,1g/ml LPS,NO (M/ml),1g/ml LPS,Cytosol/nuclear NF-B p65 fold,Cytosol Nuclear -actin,Fig. Effect of Fucoidan on the secretion of NO by LPS-stimulated RAW264.7 cells (n=3).,Fig. Effect of Fcoidan on the protein expression of NF-B p65 by LPS-stimulated RAW264.7 cells (n=3).,AA induce,Animal Model Induction of ear-skin inflammation,Fig. The effect of Fucoidan on the ear thickness of ear inflammation.,Fig. The effect of Fucoidan on RBC fraction of tissue volume of ear inflammation.,Control,Positiv