您的 Browser 不支援 Javascript , 但是並不會影響您觀看本站的內容 跳到主要內容區塊

研究報告

更新日期: 2012-01-04
點閱數: 1,984

藻藍素抗腫瘤活性之探討

年度 2006
計劃名稱 藻藍素抗腫瘤活性之探討
中文摘要 保健食品在二十一世紀佔全球食品市場之重要地位,而水產保健食品則剛碾露頭角,極具開發潛力。聚球藻(Syne chococcus sp.)屬藍綠藻,其富含藻藍(phycocyanin)為一光吸收色素-蛋白質複合體;藻藍素應用在許多營養食品補充劑。目前已知具有抗氧化能力、抗發炎、神經保護及肝臟保護作用而呈現多樣化的生理活性。本研究目的為利用濃縮收集之聚球藻,經由不同方法萃取,評估藻藍素最適之萃取條件,並將藻藍素粗萃液進一步純化,利用經純化之藻藍素,探討其抗腫瘤之活性。實驗結果顯示,於萃取條件上以凍結-解凍破碎細胞且以水萃為佳。將萃取液以超濃縮過濾濃縮約20倍後,經疏水性交換層析及陰離子交換層析純化,經Native PAGE得知其分子量約為106 KDa,再以SDS-PAGE變性裂解成兩個次單元,其分子量分別約為18.4 kDa及17.1 kDa。再者,於藻藍素穩定性方面,其於4 ℃、避光及pH=7的環境下安定性較佳。在金屬離子的影響方面,以添加濃度100 mM的HgCl2與CuSO4影響最大,其殘存率不到30%。在化學添加物方面,以添加濃度100 mM的SDS影響最大,其損失率約78.1%。至於抗腫瘤活性,C-PC對於腫瘤細胞株(Hep G2和Hep 3B)具有顯著之細胞毒性,IC50分別為30 μg/mL與70 μg/mL,而在該濃度下(30 μg/mL與70 μg/mL)對正常細胞3T3無顯著毒性。而在Hep G2細胞株中,發現以C-PC處理24小時後,其細胞型態隨著濃度的上升漸漸由平滑的角狀變成皺縮和死亡的情形。
英文摘要 Health foods (Nutraceiuticals) play the critical role on the global food market in 21st century, while the health seafood area is the new era of functional foods and exposed the great potential. Synechococcus sp., a member of cyanobacteria family, containing phycocyanin is one of the major light harvesting pigment-protein which exhibiting diverse antioxidation capacities, anti-inflammatory, neuroprotective and hepatoprotective effect. The main objectives of the study are using concentrated Synechococcus sp to investigate the optimal extraction conditions by various methodologies and evaluate the antitumor activity by employing purified phycocyanin. The results indicated that Freezing -thawing in combination with distilled water was the best method for extracting phycocyanin from Synechococcus sp.U2. The crude extracts was concentrated up to 20 fold by ultrafiltration and consecutively purified by butyl- sepharose and DEAE- sepharose resin. When analyzed by SDS-PAGE, Synechococcus sp. U2 phycocyanin migrated as two bands having an apparent molecular weight of 18.4 and 17.1 kDa. The purity extracts of 90μg/ml from Synechococcus sp.U2 scavenged DPPH 、superoxide and H202by 92.1% 、82.5% and 61.8%, respectively. The dosage of purified phycocyanin, 0.1, 0.5, and 1.0 mg/plate were used, showed no toxicity and mutagenicity. The purified phycocyanin extracts from Synechococcus sp.U2 indicated the best antimutagenic effect when the concentration of 1 mg/plate phycocyanin was employed. The purified phycocyanin extracts was stable over pH=7 at 4℃without light. In the presence of metal ions, the content of phycocyanin was reduced by adding 100 mM HgCl2 and CuSO4 and only less than 30% residual left. In related to chemical additives study, the content of phycocyanin was reduced 85.2% and 78.1% respectively by adding 100 mM HgCl2 and CuSO4. The antitumor study revealed that the phycocyanin has the effect on the viability of HepG2 and Hep3B cells, and the IC 50 for those two cells are 30 μg/ml and 70 μg/ml, respectively. However, no significant effect was observed on the viability of normal 3T3 cells. The microscopy observation indicated that the higher concentrated phycocyanin was employed, the more intensive shrinkage of HepG2 cells was found.
計畫編號 95農科-10.1.5-漁-F1(6)
網站導覽錨點