Knowledge Resource Center for Ecological Environment in Arid Area
| 唐古特白刺成分分离分析与药理活性研究 | |
| 其他题名 | Study on the separation and analysis of constituents and pharmacological activities of Nitraria tangutorum Bobr. |
| 胡娜 | |
| 出版年 | 2015 |
| 学位类型 | 博士 |
| 导师 | 索有瑞 |
| 学位授予单位 | 中国科学院大学 |
| 中文摘要 | 唐古特白刺为蒺藜科(Zygophyllaceae)白刺属(Nitraria)植物，是中国的特有种。其在柴达木地区的荒漠化治理和生态环境改善方面具有广阔的发展前景，因此具有极其重要的生态学价值。其含有较为丰富的营养成分如氨基酸、脂肪酸、矿物质元素以及多种生物活性成分如多糖、黄酮、生物碱和花色素等。白刺具有广泛的医疗价值，被用于治疗多种疾病，其具有多种的药理活性作用如降血脂、降血糖、抗氧化、抗疲劳、调节免疫和保护化学性肝损伤等作用。此外，作为一种广泛分布在柴达木地区的浆果植物，白刺也具有较高的食用价值和产品开发价值。为了更好的开发利用白刺资源，在前人研究的基础上，本文对唐古特白刺的营养成分、化学成分以及药理作用进行了较为系统的研究，主要取得了以下的研究成果和结论： 1. 以2-(7-甲基-1H-吡唑-[3,4-b] 喹啉-1-乙基)-4-甲基苯磺酸（PEB）为衍生试剂，建立了一种基于HPLC-FLD-MS技术的21种脂肪酸测定分析方法，该方法的检测限可以达到0.52-2.34ng/mL。应用该方法对不同产地、不同组织的唐古特白刺样品中的游离脂肪酸进行分析，结果表明：白刺的不同组织中均含有大量的不饱和脂肪酸，其不饱和脂肪酸所占总脂肪酸的含量均在60%以上。白刺果皮与果肉、种子、叶子中的主要不饱和脂肪酸分别是以C18:1、C18:2和C18:3为主。不同组织中所含有的饱和脂肪酸主要是以C16和C18为主。另外，不同产地的白刺相同组织中脂肪酸种类基本上没有区别，但在含量方面存在一定的差异。 2. 以10-乙基-吖啶酮-2-磺酰氯（EASC）作为衍生试剂，建立了一种于基于HPLC- FLD-MS技术的19种氨基酸的测定方法。该方法的检测限可以达到0.13-1.13nmol/L，并且在20min内实现了19种氨基酸的快速分离。应用此方法对柴达木盆地不同产地、不同组织的白刺样品中的氨基酸进行分析，结果表明：白刺的不同组织中均含有大量的氨基酸，白刺果皮与果肉中所含的氨基酸总量最高，主要是脯氨酸和色氨酸含量最高；叶子中的总氨基酸含量次之，以脯氨酸、谷氨酸和天冬氨酸为主；种子中的氨基酸总量相对最低，主要是谷氨酸和精氨酸含量为最高。另外，不同产地的白刺相同组织中氨基酸种类没有区别，但在含量方面存在一定的差异。 3. 利用响应面法优化超声辅助提取唐古特白刺种子总黄酮的工艺，即乙醇浓度66%，超声时间68min，超声温度42℃，在此最佳条件下实际测得总黄酮含量为1.20%。采用DPPH清除体系、羟基自由基清除体系和NaNO2清除体系对白刺种子中的总黄酮的抗氧化能力进行了评价，结果表明白刺种子总黄酮对多种氧化自由基均具有一定的清除作用，说明其具有一定的抗氧化活性。 4. 分别通过H2O2损伤L02细胞模型和HepG2细胞胰岛素抵抗模型来研究白刺籽油的抗氧化作用以及降血糖作用，并且与另外两种浆果籽油即枸杞籽油和沙棘籽油进行了对比研究。结果表明三种籽油均对过氧化氢损伤的L02细胞具有不同程度的预防保护作用。可以增加细胞内总蛋白含量、SOD和CAT活力，以及降低有害物质MDA的积累。白刺籽油与枸杞籽油具有一定的降血糖作用，而且枸杞籽油的降血糖作用优于白刺籽油，而沙棘籽油的降血糖作用则不明显。 5. 采用高速逆流色谱对化合物进行初步分离，结合半制备色谱技术成功的将两种非对应异构的吲哚生物碱分离出来。NMR方法鉴定两种化合物分别是(1S,3S)-1-Methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid和(1R,3S)-1-Methyl- 1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid，其纯度分别为99.9%和97.5%。其中(1R,3S)-1-Methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid为首次从本种植物中分离得到。 6. 建立大鼠高血脂模型，对白刺种子提取物、石油醚萃取部位、水部位的降血脂活性进行研究，结果表明不同剂量的各个药物组均能不同程度的降低高血脂大鼠血清中的总胆固醇、总甘油三脂、低密度脂蛋白胆固醇，提高其高密度脂蛋白胆固醇，并能在一定程度上对抗高血脂大鼠体重的异常增长。同时对各组大鼠的肝组织进行病理学切片观察也发现，各个药物组均能不同程度的改善由高血脂引起的脂肪组织和细胞形态的变化。研究发现石油醚部位的降血脂作用优于提取物，更优于水部位的降血脂的作用。 |
| 英文摘要 | Nitraria tangutorum Bobr belongs to the Nitraria genus of the Zygophyllceae, which is endemic to China. It has extremely important ecological value, because it has a broad development prospects in the governance of desertification and the ecological environment improvement of the Qaidam Basin. It contains relatively rich nutrients such as amino acids, fatty acids, mineral elements and a variety of biological active ingredients such as polysaccharides, flavonoids, alkaloid and anthocyanin, etc. This plant has broad medical value and is used in the treatment of many diseases. It has many pharmacological activities such as the effects of hypoglycemic, hypolipidemic, antioxidation, antifatigue, immune regulation and protection of chemical liver damage and so on. In addition, as a kind of berry plants widely distributed in Qaidam area, Nitraria tangutorum Bobr also has high edible value and product developmentvalue. In order to better development and utilization of this resource, a systematic study on the nutrients, chemical composition and pharmacological funcation of Nitraria tangutorum Bobr was carried out in this paper. The main research contents and results are as follows: 1. A sensitive method based on the HPLC-FLD-MS technology using 2-(7-methyl-1H-pyrazolo-[3,4-b]quinoline-1-yl)ethyl-4-methyl benzenesulfonate as precolum derivatizing reagant was developed for the analysis of fatty acids, and 21 derivatives of fatty acids were separated well. The limits of detection can be in the range of 0.52-2.34ng/mL. Free fatty acids in Nitraria tangutorum Bobr of different tissues from different regions were analyzed by this method. The result showed that different tissues of Nitraria tangutorum Bobr all contained plenty of unsaturated fatty acids, the unsaturated fatty acids could account for over 60% of the total fatty acids. The main unsaturated fatty acids in sarcocarps, seeds and leaves were mainly C18:1, C18:2 and C18:3, respectively. The saturated fatty acids in all tissues were mainly C16 and C18. In addition, there were no differences in the type of fatty acid of Nitraria tangutorum Bobr with the same tissues from different regions, but there were some differences in the content of fatty acids. 2. A sensitive method based on the HPLC-FLD-MS technology using 10-ethyl-acridong-2-sulfonyl chloride (EASC) as precolum derivatizing reagent was developed for the analysis of amino acids, and 19 derivatives of amino acids were separated well within 20min. The limits of detection could be in the range of 0.13-1.13nmol/L. Amino acids in Nitraria tangutorum Bobr of different tissues from different regions were analyzed by this method. The result showed that different tissues of Nitraria tangutorum Bobr all contained plenty of amino acids. The content of total amino acids in the sarcocarps was the highest, and the content of the total amino acids in seeds was the lowest. The mian amino acids in sarcocarps, seeds and leaves were Pro and Trp, Pro, Glu and Asp, and Glu and Arg, respectively. In addition, there were no differences in the type of amino acid of Nitraria tangutorum Bobr with the same tissues from different regions, but there were some differences in the content of amino acids. 3. The ultrasound-assisted extraction process parameters of total flavonoids from Nitraria tangutorum Bobr seeds were optimized with a response surface methodology (RSM). The optimum extraction conditions were as follows: 65% ethanol, extraction time 68min, and extraction temperature 42℃. Under the conditions, the optimum extraction ratio of the total flavonoids from Nitraria tangutorum Bobr seeds was 1.20%. 1,1-Diphenyl-2-picrylhydrazyl(DPPH) radical-scavenging assay, nitrite (NaNO2) scavenging assay and hydroxyl (?OH) radical-scavenging assay were used to evaluate the vitro antioxidant activity of total falvonoid, and the result showed that total falvonoids of Nitraria tangutorum Bobr seeds exhibited a variety of oxide free radical scavenging effect. 4 |
| 中文关键词 | 唐古特白刺 ; 成分 ; 分离 ; 分析 ; 药理活性 |
| 英文关键词 | Nitraria tangutorum Bobr ingredient separation analysis pharmacological activities |
| 语种 | 中文 |
| 国家 | 中国 |
| 来源学科分类 | 植物学 |
| 来源机构 | 中国科学院西北高原生物研究所 |
| 资源类型 | 学位论文 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/287586 |
| 推荐引用方式 GB/T 7714 | 胡娜. 唐古特白刺成分分离分析与药理活性研究[D]. 中国科学院大学,2015. |
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