The Induction of NO and Anti-tumor Effects of Extracted and Purified Polysaccharides from Talinum triangulare

• Main Authors: Jiang, Bohao, 蔣博豪
• Other Authors: Tsai, Mingshium
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/11362057824577181527

碩士 === 大葉大學 === 生物產業科技學系 === 100 === According to the statistic results showed by Department of Health, cancer has been the first cause of death in the past three decades. At present, most anti-cancer drugs are less specific, and they may cause many side-effects. Therefore, extracts or compounds from herbals or natural plants have been good choice for evaluation of anti-cancer therapy. Some health-improving activities of Talinum triangulare are widely known and spread, but only anti-oxidative activity and hepatoprotective effect of extracted polysaccharides from Talinum triangulare have been reported. In this study, we examined whether the water-extracted polysaccharides (TTP) from Talinum triangulare can promotion of inflammation and have anti-tumor activities. We also try to explore their active polysaccharide ingredients and mechanisms of inflammation. We first used orthogonal array L9 to find the optimal extraction conditions of TTP by heat reflux and ultrasonic extractions, respectively. The optimal heat reflux extraction conditions are 95℃ incubation temperature, 1:20 ratio of material and water, and 150 minutes of incubation time. The optimal ultrasonic extraction conditions are 85℃ incubation temperature, 1:20 ratio of material and water, and 75 minutes of incubation time. Both methods obtain the maximum 3.1% conversion yield. After ethanol precipitation, TTP were re-dissolved in distilled water. TTP was separated into two molecular weight ranges by dialysis. Various amounts of two kinds of TTP were added into media of RAW264.7 macrophages for analyzing their inflammation activities by measuring nitric oxide (NO) productions. We found that TTP with molecular weights larger than 14kDa could induce more NO synthesis than TTP with molecular weights lower than 14kDa. This result suggests that the larger TTP have higher inflammation activity than the smaller one. Because the NO production is one of inflammatory responses that can suppress tumor cell growth, we analyzed the anti-tumor activity of TTP treated RAW264.7 cells. We collected the media of RAW264.7 cells pre-treated with two types of TTP, virespectively. We treated human cervical cancer HeLa and colorectal cancer SW620 cells with these media. We found the media of TTP pre-treated RAW264.7 cells could significant inhibit HeLa and SW620 cells, and the larger TTP has higher inhibition activity than the smaller TTP. However, both types of TTP have no direct anti-tumor activity. Furthermore, we separated and purified seven kinds of polysaccharides from the large TTP by anion-exchange chromatography, and then we analyzed their inflammation and anti-tumor activities, respectively. Results showed that each separated polysaccharides from TTP has less NO induction and lower anti-tumor activities than the original large TTP. These results may account for that the inflammation and anti-tumor activities are not induced by single type of polysaccharide. From our results, we conclude that TTP has anti-tumor activities through regulating the functions of macrophages.