花色苷类物质抗氧化生理活性的研究
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摘要
活性氧可对生物体产生一系列的有害作用，例如进攻组织内的多不饱和脂肪酸，蛋白质和核酸等，造成氧化损伤，而这种氧化损伤与很多衰老疾病如心脏病、神经系统紊乱和癌症等的病因有关。
花色苷是一种在植物中普遍存在的酚类物质，是很多植物叶花和果实呈现鲜艳颜色的原因。它具有抑制活性氧的生成和清除活性氧的生理活性，花色苷的抗氧化性源于其结构中的侧环酚羟基。
本文研究了三种富含花色苷的农产品——黑豆，黑米和紫甘蓝中花色苷类物质的溶剂浸提法的提取条件和采用X-5吸附树脂柱层析法的初步精制条件。采用国标法对制备和得到的五种花色苷样品的色价进行了测定，测定结果均大于40，并通过纸层析法和紫外可见光谱法初步推测了样品的可能花色苷成分。
采用没食子酸等价法确立了黑豆，黑米，紫甘蓝，紫甘薯和萝卜花色苷样品的总酚含量与其水溶液可见光区最大吸收波长下吸光度的关系，为体外抗氧化实验提供了定量基础。以下分别采用还原K_3Fe(CN)_6法考查五种花色苷样品的还原力差别、采用邻二氮菲(1，10-phenanthroline)-Fe~(2+)比色法考查样品清除羟自由基能力和采用Fe~(2+)引发卵磷脂脂质体氧化-TBA比色法考查样品抗脂质体氧化的能力，得到的总体结果为：黑米、黑豆花色苷强于紫甘薯花色苷，萝卜花色苷次之，紫甘蓝花色苷相对较弱。同时，依据文献，也对花色苷的结构对其抗氧化能力的影响进行了分析。
最后，试图以昆明种小鼠通过喂饲高脂饲料建立一种高血脂动物模型，采用组间对照的方法，通过测定血清生化指标血清总甘油三脂(TG)，血清总胆固醇(TC)，高密度脂蛋白胆固醇含量(HDL-c)，计算动脉硬化指数(A.I.)，评估黑豆花色苷的抗动脉硬化的能力。由于在实验中各组实验动物中有很多小鼠死亡，导致阳性对照组和给药Ⅰ组的数量严重不足，无法估计所选择的高脂饲料是否成功建立小鼠的高血脂模型。从而无从评估黑豆花色苷抗动脉硬化能力。
Reactive oxygen species (ROS) are very damaging to organisms, since they can attack polyunsarural fatty acids (PUFA), proteins and nuclear acid in tissues to induce oxidation. This oxidative damage is considered to play a causative role in aging and degenerative diseases associated with it, such as heart disease, cognitive dysfunction and cancer.
Anthocyanins are polyphenols widely distributed in plants, and they contribute to the brilliant color of blue, red, scarlet in leaves, flowers and fruits. It is reported that they have potentials inhibiting the formation of ROS and scavenging ROS, and the antioxidant ability due to the hydroxyl groups substituted in the aromatic B-ring.
Optimum extract conditions of anthocyanins from black bean, black rice and purple cabbage and purification conditions of X-5 absorbent resin column chromatography were studied. The colorities of five anthocyanins samples were determined with GB method, all were higher than 40. Possible anthocyanin components of these five samples were tentatively inferred from paper chromagraphy and UV-Visible spectrum .
After having obtained the relationship between the phenol content and the samples' absorbance at maximum absorption point of visible spectrum, three systems were adopted to assess the reducing power, scavenging ability on hydroxyl radical, inhibition ability on oxidation, induced by Fe2+, of lecithin liposome system of anthocyanins. Antioxidant abilities of anthocyanins extracted from black bean and black rice were stronger than the ones from purple sweet potatoes, red-core radishes and purple cabbage.
A model of Kunming rats with hyperlipidemia resulting from high-fat feeding was tried to found to determine the anti-atherogenic ability of anthocyanins from black bean.
花色苷是一种在植物中普遍存在的酚类物质，是很多植物叶花和果实呈现鲜艳颜色的原因。它具有抑制活性氧的生成和清除活性氧的生理活性，花色苷的抗氧化性源于其结构中的侧环酚羟基。
本文研究了三种富含花色苷的农产品——黑豆，黑米和紫甘蓝中花色苷类物质的溶剂浸提法的提取条件和采用X-5吸附树脂柱层析法的初步精制条件。采用国标法对制备和得到的五种花色苷样品的色价进行了测定，测定结果均大于40，并通过纸层析法和紫外可见光谱法初步推测了样品的可能花色苷成分。
采用没食子酸等价法确立了黑豆，黑米，紫甘蓝，紫甘薯和萝卜花色苷样品的总酚含量与其水溶液可见光区最大吸收波长下吸光度的关系，为体外抗氧化实验提供了定量基础。以下分别采用还原K_3Fe(CN)_6法考查五种花色苷样品的还原力差别、采用邻二氮菲(1，10-phenanthroline)-Fe~(2+)比色法考查样品清除羟自由基能力和采用Fe~(2+)引发卵磷脂脂质体氧化-TBA比色法考查样品抗脂质体氧化的能力，得到的总体结果为：黑米、黑豆花色苷强于紫甘薯花色苷，萝卜花色苷次之，紫甘蓝花色苷相对较弱。同时，依据文献，也对花色苷的结构对其抗氧化能力的影响进行了分析。
最后，试图以昆明种小鼠通过喂饲高脂饲料建立一种高血脂动物模型，采用组间对照的方法，通过测定血清生化指标血清总甘油三脂(TG)，血清总胆固醇(TC)，高密度脂蛋白胆固醇含量(HDL-c)，计算动脉硬化指数(A.I.)，评估黑豆花色苷的抗动脉硬化的能力。由于在实验中各组实验动物中有很多小鼠死亡，导致阳性对照组和给药Ⅰ组的数量严重不足，无法估计所选择的高脂饲料是否成功建立小鼠的高血脂模型。从而无从评估黑豆花色苷抗动脉硬化能力。
Reactive oxygen species (ROS) are very damaging to organisms, since they can attack polyunsarural fatty acids (PUFA), proteins and nuclear acid in tissues to induce oxidation. This oxidative damage is considered to play a causative role in aging and degenerative diseases associated with it, such as heart disease, cognitive dysfunction and cancer.
Anthocyanins are polyphenols widely distributed in plants, and they contribute to the brilliant color of blue, red, scarlet in leaves, flowers and fruits. It is reported that they have potentials inhibiting the formation of ROS and scavenging ROS, and the antioxidant ability due to the hydroxyl groups substituted in the aromatic B-ring.
Optimum extract conditions of anthocyanins from black bean, black rice and purple cabbage and purification conditions of X-5 absorbent resin column chromatography were studied. The colorities of five anthocyanins samples were determined with GB method, all were higher than 40. Possible anthocyanin components of these five samples were tentatively inferred from paper chromagraphy and UV-Visible spectrum .
After having obtained the relationship between the phenol content and the samples' absorbance at maximum absorption point of visible spectrum, three systems were adopted to assess the reducing power, scavenging ability on hydroxyl radical, inhibition ability on oxidation, induced by Fe2+, of lecithin liposome system of anthocyanins. Antioxidant abilities of anthocyanins extracted from black bean and black rice were stronger than the ones from purple sweet potatoes, red-core radishes and purple cabbage.
A model of Kunming rats with hyperlipidemia resulting from high-fat feeding was tried to found to determine the anti-atherogenic ability of anthocyanins from black bean.
引文
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[2] 赵保路．氧自由基和天然抗氧化剂[M]．北京：科学出版社．1999．7
[3] Pier-Giorgio Pietta. Flavonoids as Antioxidants [J]. J. Nat. Prod., 2000,63:1035-1042
[4] 郑晶泉．抗氧化剂抗氧化实验研究进展[J]．国外医学卫生分册．2000，27(1)：37-40
[5] 林怡玲，邱莉莉，吴懿哲，萧明熙．FreeRadicals, Atherosclerosis, and Development of Antioxidants as Antiatherogenic Agents [J]．食品科学(台湾)．1997，24(6)：690-705
[6] 金宗濂，文镜，唐粉芳，陈文．功能食品评价原理及方法[M]．北京：北京大学出版社．1995，10
[7] 上海第一医学院．医用生物化学[M]．北京：人民卫生出版社．1979，3
[8] 于瑞敏．抗氧化剂与动脉粥样硬化[J]．中国动脉硬化杂志．1995，3(3)：267-270
[9] Palinski W, rosenfeid ME, Yia-Hertruala S, et al. Probucol inhibits oxidative modification in vivo[J]. Proc Natl Acad Sci USA. 1989, 86:13 72-1376
[10] Bruce N. Ames. Dietary Carcinogens and Anticarcinogens[J]. Science. 1983, 221: 1256-1264.
[11] 马自超，庞业珍．天然色素化学及生产工艺学[M]．北京：中国林业出版社．1994
[12] J．B．哈本等编，戴伦凯，谢玉如等译．黄酮类化合物[M]．北京：科学出版社．1983，4．
[13] 蔡正宗，陈叔凰．黑糯米中两种主要花色素苷之结构鉴定[J]．食品科学(台湾)．1996，23(3)：444-452．
[14] 杨正宪，蔡正宗．桑葚之Anthocyanins结构鉴定[J]．食品科学(台湾)．1994，21(5)：319-330．
[15] Johanna Bakker, Colin F. Timberlake. Isolation, Identification, and Characterization of New Color-Stable Anthocyanins Occurring in Some Red Wines [J]. J. Agric. Food Chem. 1997, 45:35-43
[16] Victor Hong, Ronald E. Wrolstad. Characterization of Anthocyanins-containing Colorants and Fruits Juices by HPLC/Photodiode Array Detection [J]. J. Agric. Food Chem. 1990, 38(3): 698-708
[17] Victor Hong, Ronald E. Wrolstad. Use of HPLC Separation/Photodiode Array Detection for Characterization of Anthocyanins[J]. J. Agric. Food Chem. 1990, 38(3): 708-715
[18] 张红雨，陈德展．酚类抗氧化剂清除自由基活性的理论表征与应用[J]．生物物理学报．2000，16(1)：1-9
[19] Yasuko Noda, Takao Kneyuki, Kiharu Igarashi et al. Antioxidant activity of nasunin, an anthocyanin in eggplant peels [J]. Toxicology. 2000, 148: 119-123.
[20] 龙盛京．化学发光法测定黑米色素抗氧化作用[J]．分析实验室．1999，4：76-79
[21] 孙玲．黑米的抗氧化性及其黄酮和种皮色素的关系[J]．2000，3：246-249
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