Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/50934
標題: Antioxidant and anti-inflammatory effects of pine (Pinus morrisonicola Hayat) needles
台灣五葉松松針之抗氧化及其抗發炎效果
作者: Huang, Din-Wen
黃鼎文
關鍵字: 台灣五葉松;P. morrisonicola Hay.;抗氧化性;LDL;類黃酮;多酚類;抗發炎;antioxidant;LDL;inflammatory;LPS;NO;ROS;iNOS;COX-1;COX-2;IL-1
出版社: 食品科學系
摘要: 
摘 要
本研究乃探討台灣五葉松松針之抗氧化性、抑制銅離子誘導人體LDL氧化、抗發炎以及其具生物活性之成分。結果顯示,台灣五葉松松針經不同極性溶劑 (正己烷、乙酸乙酯、甲醇、水) 萃取後,以乙酸乙酯區分層 (NEE)抗氧化性最好,其清除DPPH自由基之IC50為80 μg/ mL;500 μg/mL之作用劑量,在TEAC評估系統中即相當於 357 μM Trolox,在ORAC中亦相當於25.7 μM之Trolox,對於超氧陰離子(O2•-)及一氧化氮(NO)則分別有60%及25%之清除效果。Cu2+/LDL氧化修飾系統下,NEE具有延緩LDL共軛雙烯(conjugated diene, CD)、脂質過氧化物 (TBARS)生成,以及減低LDL相對電泳移動量 (related electrophoresis movement, REM)的現象。同時,對於巨噬細胞RAW 264.7吞噬氧化LDL (ox-LDL)作用上,亦具有顯著之抑制效果。分析其各區分層之抗氧化成分,NEE具高總多酚 (123 mg/g)及類黃酮 (64 mg/g)含量。進一步以HPLC分析其多酚化合物種類,結果顯示,NEE含有epicatechin(EC)及p-coumaric acid。因此,NEE之高量多酚類成分可能為其表現較高抗氧化性的原因。
以LPS誘導RAW 264.7巨噬細胞,模擬體內發炎及NO過度生成之狀態。結果顯示,NEE能有效降低LPS誘發RAW 264.7細胞胞內活性氧及NO上升的狀況,作用劑量達75 μg/mL有最佳之表現。探討其抗發炎分子機制,NEE可有效降低COX-2、iNOS及IL-1之mRNA表現,以Western bloting分析上述基因之蛋白質表現亦得到類似的抑制結果。此顯示,NEE可藉由調節前發炎激素IL-1及發炎相關酵素iNOS及COX-2之表現情形,以降低體內發炎之狀態。
綜上所述,NEE可以有效清除自由基降低氧化壓力,減緩LDL
氧化,且NEE藉由抑制iNOS與COX-2之表現調節發炎反應。

Abstract
The aims of this study were to investigate the antioxidant effect, inhibition of copper-mediated LDL oxidation, anti-inflammatory activity of pine (P. morrisonicola Hay.) needles and to analyze its polyphenol contents. Pine needles were extracted with four solvents (hexane, ethyl acetate, methanol and water). Results showed that ethyl acetate extracts of pine needles (NEE) had good antioxidant activity in DPPH radical scavenging capacity (IC50= 80 μg/mL), TEAC and ORAC values (500 μg/mL NEE equivalent 357 and 25.7 μM Trolox, respectively). Scavenging effects of NEE on super-oxide anion and nitric oxide radical were 60% and 25%, respectively, at a concentration of 500 μg/mL. In the Cu2+/LDL oxidative modification system, NEE significantly retarded LDL oxidation through prolonging the lag phase of conjugated diene formation, decreasing the amounts of lipid peroxidation products (TBARS) and slowing the relative electrophoretic mobility (REM) of LDL. In addition, NEE significiently inhibited RAW 264.7 murine macrophages uptaking ox-LDL. As to antioxidant contents of various pine extracts, NEE has the high amounts of polyphenol (123 mg/g) and flavonoids (64 mg/g) content. A HPLC method was used to indentify and quantify the polyphenol contents in NEE, the results showed that NEE contained epicatechin (EC) and p-coumaric acid. Thus, NEE showed higher antioxidant activity than other solvents extracts might be attributed to its abundant polyphenol contents.
Nitric oxide (NO) production had been implicated in the process of
carcinogenesis and inflammation. In this study, RAW 264.7 macrophage
cells were treated with LPS to simulate the inflammatory model in vitro. The results showed that both NO and ROS were decreased after treating with NEE at the concentration of 75 μg/mL. NEE inhibited the protein and mRNA expression of COX-2, iNOS and IL-1in RAW 264.7 macrophagesIt is suggested that the anti-inflammatory activity of NEE was at least, partly due to their inhitory effects on the activation of iNOS, COX-2 and IL-1.
In conclusion, NEE could attenuate the oxidative stress by scavenging free radicals; reduce excessive iNOS and COX-2 expression in macrophages and lipoprotein oxidation in vitro.
URI: http://hdl.handle.net/11455/50934
Appears in Collections:食品暨應用生物科技學系

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