十五種中藥材抗氧化與抗發炎能力評估

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標題:	十五種中藥材抗氧化與抗發炎能力評估 Evaluation of antioxidant and anti-inflammatory of fifteen kinds of Chinese herbal medicine
作者:	胡筱婷
貢獻者:	王柏森黃明星嘉南藥理科技大學：藥物科技研究所
關鍵字:	連翹旋覆花蔓荊子抗氧化抗發炎 HPLC Forsythia suspensa antioxidative HPLC anti-inflammation Vitex trifolia Inula britannica
日期:	2010
上傳時間:	2010-12-30 15:34:01 (UTC+8)
摘要:	近年來，隨著國人健康意識逐漸抬頭，中草藥植物的開發與運用，已成為重要的發展趨勢。本研究從數十種中草藥中，以總多酚含量測定及總黃酮含量測定篩選出富含多酚及類黃酮的材料，分別為女貞子、升麻、牛蒡子、地骨皮、辛夷、防風、知母、羌活、金銀花、荊芥、旋覆花、連翹、紫蘇、蒼耳子、蔓荊子等十五種實驗材料。本研究分為試管內與試管外試驗，第一部分探討其DPPH自由基清除率、TEAC總抗氧化能力、還原力、亞鐵離子螯合能力、微脂粒（liposome）氧化作用、Superoxide自由基清除能力、NO抑制能力、酪胺酸酶活性分析、總多酚含量、類黃酮含量測定等九個抗氧化平台找出較具潛力之藥材，進而建立此類藥材之HPLC化學成份指紋圖譜。第二部分為藉由巨噬細胞(RAW264.7)模式系統來評估中草藥萃取物對LPS誘導一氧化氮(nitric oxide, NO)生成率及分析胞內活性氧 (ROS)含量。研究結果顯示，在抗氧化分析方面，發現連翹、旋覆花及蔓荊子有最佳的DPPH自由基清除率，連翹、旋覆花在濃度為100 ppm及蔓荊子在濃度為150 ppm時均超過90 %；TEAC值測定結果顯示連翹、旋覆花在濃度為75 ppm及蔓荊子在濃度為100 ppm時均超過90 %；亞鐵離子螫合能力結果顯示連翹、旋覆花在濃度為600 ppm及蔓荊子在濃度為700 ppm亞鐵離子螫合能力達80％以上；在還原能力測定結果顯示連翹、旋覆花在濃度為100 ppm及蔓荊子在濃度為150 ppm時其OD700值均超過0.477以上；在微脂粒氧化作用之抑制力試驗中，連翹、旋覆花於濃度為400 ppm時，抑制效果分別為93.3%、89.8%，蔓荊子在濃度600 ppm時抑制效果為95.9%；在清除superoxide方面連翹在濃度400 ppm，旋覆花、蔓荊子在濃度600 ppm時均超過80 %；在抑制NO方面連翹、旋覆花及蔓荊子在濃度250 ppm時抑制效果分別為94.3%、90.8%、88.6%；在總多酚定量試驗，連翹、旋覆花及蔓荊子每克萃取物含有相當於 226.5 mg、210.7 mg、139.9 mg之gallic acid。總類黃酮測定，連翹、旋覆花及蔓荊子之每克萃取物含有相當於126.4mg、125.2 mg、82.1mg之rutin。酪胺酸酶活性評估顯示，連翹、旋覆花及蔓荊子在濃度1500 ppm、2000 ppm與2000 ppm時，抑制率分別為87.0%、90.3% 與88.4%。連翹、旋覆花及蔓荊子HPLC的指紋圖譜，顯示含有quercitrin、rutin 、chlorogenic acid 、 kaempferol、 vanillic acid、 luteolin, ferulic acid等成份。在抗發炎試驗方面旋覆花在濃度100ppm 時有超過50%的抑制效果；連翹在100 ppm的時有超過50%的抑制效果；女貞子、防風、紫蘇葉、金銀花在濃度400ppm時抑制效果均超過50%。在ROS試驗方面，有四種樣品具有抑制ROS生成效果，分別是金銀花、羌活、知母以及連翹。在LPS誘導細胞產生ROS方面有七種樣品具有抑制生成效果，分別是金銀花、羌活、知母、升麻、蒼耳子、牛蒡子以及荊芥具有抑制ROS生成效果。 In recent years, people pay more attention on the health issues, therefore, the development and application of the Chinese herbals have become the world trend. In this study, 15 Chinese herbal medicine water extracts including Schizonepeta tenuifolia, Vitex trifolia, Saposhnikovia divaricata, Forsythia suspensa, Lonicera japonica, Inula britannica, Anemarrhena asphodeloides, Cimicifuga heracleifolia, Lycium chinense, Arctium lappa, Ligustrum lucidum, Perilla frutescens, Xanthium strumarium, Notopterygium incisum and Magnolia biondii were conducted to determine their antioxidant and anti-inflammatory effects. In Part 1, we focused on the studies of the antioxidant activities, include the scavenging effect on α,α-diphenyl-β-picrylhydrazyl(DPPH) radicals, the trolox equivalent antioxidant capacity (TEAC), reducing power, ferrous ion chelating power, inhibition NO capacity, scavenging superoxide radical capacity and inhibitory effect on lipid peroxidation total polyphenols and flavonoids. The HPLC-UV finger print chromatogram and antityrosinase effects of the potent herbs were also measured. In part two, we investigated the effects of Chinese herbals, on RAW 264.7 macrophages production NO induced by LPS. The results show that all these herbal extracts demonstrate antioxidant abilities and in dose-dependent manner. Forsythia suspensa, Inula britannica, and Vitex trifolia have the greatest DPPH radical scavenging effect. Inula britannica, Vitex trifolia and Cuscuta chinessis have better antioxidative activity by TEAC. The ferrous ion chelating capacity of the Forsythia suspensa, Inula Britannica and Vitex trifolia have the best effect. The reducing power of the Forsythia suspensa, Inula Britannica and Vitex trifolia have the better effect. Inhibition NO capacity of Forsythia suspensa, Inula Britannica and Vitex trifolia have the better inhibition effect. Scavenging superoxide radical capacity of the Forsythia suspensa, Inula Britannica and Vitex trifolia have the greatest superoxide radical scavenging effect. Forsythia suspensa, Inula Britannica and Vitex trifolia have the better effect of inhibitory effect on lipid peroxidation. Forsythia suspensa, Inula Britannica and Vitex trifolia had total polyphenol contents (226.5 mg、210.7 mg and 139.9mg of gallic acid equivalent, respectively) and total flavonoid contents (126.4 mg、125.2mg and 82.1mg equivalent of rutin, respectively). Forsythia suspensa, Inula britannica and Vitex trifolia exhibit potent inhibitory effects on tyrosinase. A high performance liquid chromatography analysis revealed that phenolic acids and flavonoids, such as quercitrin, rutin, chlorogenic acid, kaempferol, vanillic acid, luteolin and ferulic acid . We found that six extracts of Chinese herbs could inhibit nitric oxide generation induced by LPS by 50% or more in RAW 264.7 cells at a concentration of 400µg/ml including Inula britannica, Forsythia suspensa, Saposhnikovia divaricata, Ligustrum lucidum, Perilla frutescens and Lonicera japonica. Inula britannica has the best inhibitory effect. We found that four extracts of Chinese herbs could inhibit ROS production of RAW 264.7 cell including Lonicera japonica, Notopterygium incisum, Anemarrhena asphodeloides and Forsythia suspense. That seven extracts of Chinese herbs could inhibit ROS production generations induced by LPS in RAW 264.7 cell including Schizonepeta tenuifolia, Anemarrhena asphodeloides, Cimicifuga heracleifolia, Arctium lappa, Lonicera japonica, Xanthium strumarium and Notopterygium incisum.
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