Antioxidant Activity of Total Flavonoids Extracts from Folium of Artemisiae argyi in Vitro and in Vivo
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摘要: 本文探究了艾叶总黄酮提取物（total flavonoids extract from folium of Artemisia argyi，TFEFAA）主要成分含量及其体内外抗氧化活性。本文采用高效液相色谱（HPLC）法同时检测TFEFAA中异泽兰黄素和棕矢车菊素的含量；以1，1-二苯基-2-三硝基苯肼（DPPH）、2，2-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐（ABTS）和羟自由基三种自由基的清除率和总还原力为指标，考察TFEFAA的体外抗氧化活性；将秀丽隐杆线虫（线虫）同期化后分别培养在含有0、50、100 μg/mL的TFEFAA的线虫生长培养基中，研究TFEFAA对线虫热应激和H2O2氧化应激的影响，同时检测线虫体内相关抗氧化酶活力和丙二醛（MDA）的含量。结果表明，异泽兰黄素和棕矢车菊素的含量分别为0.2%和0.06%；TFEFAA对DPPH自由基、ABTS+自由基和羟自由基均具有显著的清除作用（P<0.05），其半数清除浓度（IC50）分别为96.51、67.73和279.38 μg/mL，清除能力与TFEFAA均呈剂量-效应关系，同时还具有较强的总还原力；100 μg/mL TFEFAA能极显著延长线虫在热应激和H2O2氧化应激条件下的存活时间（P<0.01），同时极显著提高线虫体内超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的活力（P<0.01），并且显著降低线虫体内MDA含量（P<0.05）。TFEFAA在体内外均具有较好的抗氧化活性，其机制可能与提高体内抗氧化酶活性有关。因此，TFEFAA有望被开发成新型天然绿色食品抗氧化剂。Abstract: To explore the content of the main components of the total flavonoids extract from folium of Artemisia argyi(TFEFAA) and its antioxidant activity in vivo and in vitro. In this paper, high-performance liquid chromatography(HPLC) was used to simultaneously determine the content of eupatilin and jaceosidin in TFEFAA, the antioxidant activity of TFEFAA in vitro was investigated with the scavenging rate of 1, 1-diphenyl-2-trinitrophenylhydrazine(DPPH), 2, 2-diazo-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt(ABTS) and hydroxyl radical and total reducing power as indexes. Caenorhabditis elegans(C. elegans) were cultured in nematode growth media containing 0, 50 and 100 μg/mL TFEFAA respectively. The effects of TFEFAA on heat stress and H2O2 oxidative stress of C. elegans were studied. At the same time, the related antioxidant enzyme activity and malondialdehyde(MDA) content in C. elegans were detected. The results showed that the contents of eupatilin and jaceosidin were 0.2% and 0.06% respectively and TFEFAA had a significant scavenging effect on DPPH radical, ABTS+ radical and hydroxyl radical, and its IC50 were 96.51, 67.73 and 279.38 μg/mL, respectively. The scavenging ability had a dose-effect relationship with TFEFAA, and it also had a strong total reducing power. 100 μg/mL TFEFAA can significantly prolong the survival time of nematodes under heat stress and H2O2 oxidative stress(P<0.01), at the same time significantly increase the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) in nematodes(P<0.01), and significantly reduce the MDA contents in nematodes(P<0.05). TFEFAA had good antioxidant activity in vivo and in vitro, and its mechanism may be related to improving antioxidant enzyme activity in vivo. Therefore, TFEFAA would be expected to be developed into a new natural green food antioxidant.
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