治疗高尿酸血症的中药筛选及作用机制研究
文献类型：学位论文
| 作者 | 徐晨 |
| 学位类别 | 博士 |
| 答辩日期 | 2015-04 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 中国科学院长春应用化学研究所 |
| 导师 | 刘志强 |
| 关键词 | 高尿酸血症 超氧阴离子 黄嘌呤氧化酶抑制剂 代谢组学 中药 |
| 中文摘要 | 高尿酸血症（hyperuricemia）是一种嘌呤代谢障碍疾病，近年来发病率迅速增加。目前，安全、有效、经济、副作用小的高尿酸血症治疗药物较少。我国有丰富的中药资源，中药安全有效，一直是新药研发的重要来源，从中药资源筛选治疗高尿酸血症的药物并研究其体内作用机制具有重要的意义。 本研究首先建立了一种筛选超氧阴离子清除剂的新方法并测定了多种中药的活性；还运用离心超滤质谱法筛选了中药复方二妙丸中抑制黄嘌呤氧化酶的活性成分；结合课题组前期实验结果，对二妙丸、黄柏、苍术、卷柏和虎杖治疗高尿酸血症大鼠的整体作用机制进行代谢组学研究并将主要代谢物进行定量，阐明了以上中药治疗高尿酸血症大鼠的作用机制。主要结果概况如下： 1. 建立一种测定超氧阴离子清除能力的方法，该法简单、快速、灵敏度高。我们运用此法测定了生物样品和中药提取物的超氧阴离子清除能力。在所测的27种中药中，五味子的超氧阴离子清除能力最好，银杏叶、刺五加叶、知母、山茱萸、葛根、黄芩和山楂叶也都是很好的超氧阴离子清除剂。 2. 采用离心超滤质谱技术从中药复方二妙丸中筛选并鉴定出9种与黄嘌呤氧化酶相互作用的化合物，分别为黄柏碱、木兰花碱、莲心季铵碱、蝙蝠葛仁碱、巴马汀、非洲防己碱、药根碱、小檗红碱及小檗碱。其中巴马汀、小檗红碱、小檗碱与黄嘌呤氧化酶作用较强，结合强度均在10%以上。 3. 采用基于超高效液相色谱-四级杆-飞行时间质谱仪（UPLC-Q-TOF/MS）联用技术的代谢组学方法研究中药治疗高尿酸血症大鼠的作用机制。运用主成分分析（PCA）、偏最小二乘法-判别分析（PLS-DA）和正交偏最小二乘法-判别分析（OPLS-DA）的方法对正常大鼠、高尿酸血症模型大鼠和给药组（二妙丸组、黄柏组、苍术组、卷柏组和虎杖组）大鼠尿中的内源性代谢物进行分析，寻找并鉴定了与中药治疗高尿酸血症相关的潜在生物标记物。 4. 采用超高效液相色谱-三级四极杆质谱仪（UPLC-TQ-MS）联用法定量健康组、模型组和给药组大鼠尿液中13个主要潜在生物标记物包括黄尿酸、犬尿酸、吲哚-3-羧酸、肌酐、胞嘧啶、马尿酸、尿苷、硫酸吲哚酚、辛二酸、癸二酸、尿囊素、3-羟基-2-氨基苯甲酸和尿酸。所建方法灵敏、快速、重现性好、准确度高。定量结果表明，第四章中筛选出的潜在生物标记物准确可靠；二妙丸对高尿酸血症大鼠的治疗作用好于其单味药黄柏和苍术，且苍术比黄柏药效要好；虎杖对高尿酸血症大鼠的治疗作用优于卷柏。 |
| 英文摘要 | Hyperuricemia is a disease associated with the disfunction of purine metabolism. The incidence of hyperuricemia has increased rapidly in recent years. And there are few drugs which are safe, effective, economical and low side effect. China is rich of traditional Chinese medicine (TCM) which is safe and effective and is an important sorce of new drugs. It is meaningful to screen of drugs for treating hyperuricemia from TCM and study the mechanism of action of TCM. We have developed a new analytical method to screen of superoxide radical anion scavengers and measured the activities of various TCMs. We also have screened of xanthine oxidase inhibitors from TCM formula Ermiao wan using ultrafiltration UPLC-MSn. At last, metabonomics studies of Ermiao wan, Phellodendri cortex, Atractylodis rhizome, Selaginella tamariscina and Polygonum cuspidatum for hyperuricemia in rats were conducted and the main metabolites were quantified to clarify the mechanism of action of the TCM. The main results are summarized as follows: 1. A novel analytical method for measuring O2?? scavenging activity was developed. The method was quite simple, rapid and sensitive. The method has been successfully employed for measuring O2?? scavenging activities of biological and herbal extracts. Among the 27 TCMs, Schisandra chinensis showed the best O2?? scavenging activity. Folium ginkgo, Folium acanthopanacis senticosi, Anemarrhena asphodeloides, Cornus officinalis, Radix puerariae, Radix scutellariae and Folium crataegi were also good O2?? scavengers. 2. An ultrafiltration UPLC/Q-TOF/MS assay was used to screen of xanthine oxidase inhibitors in traditional Chinese medicine formula Ermiao wan. Nine compounds were screened and identified as potential xanthine oxidase inhibitors in Ermiao wan. The nine compounds were phellodendrine, magnoflorine, lotusine, menisperine, palmatine, columbamine, jatrorrhizine, berberrubine and berberine. Among them, palmatine, berberrubine and berberine had higher activities with binding degree larger than 10%. 3. A urinary metabonomics study based on ultra-performance liquid chromatography-electrospray ionization quadruple time-of-flight/mass spectrometry (UPLC-ESI-QTOF/MS) coupled with pattern recognition approach was conducted to study the mechanism of TCM in treating hyperuricemic rats. The significant differences in metabolic profiles were observed from drug-treated group, model group and healthy control group by using principal components analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least-squares discriminant analysis (OPLS-DA). Several metabolites were identified as potential biomarkers in response to the therapeutic effects of TCM. 4. A targeted quantitative approach based on ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC–TQ-MS) was applied to quantify the 13 main potential biomarkers including cytosine, creatinine, xanthurenic acid, kynurenic acid, hippuric acid, indole-3-carboxylic acid, allantoin, uric acid, uridine, 3-Hydroxyanthranilic acid, indoxyl sulfate, suberic acid and sebacic acid in the urine of healthy control group, model group and drug-treated group. The method was sensitive, rapid and accurate. The results showed that the potential biomarkers screened in Chapter 4 were reliable. What’s more, Ermiao wan had better therapeutic effect on hyperuricemic rats than Phellodendri cortex and Atractylodis rhizome. And Atractylodis rhizome was more effective than Phellodendri cortex. Polygonum cuspidatum had better therapeutic effect on hyperuricemic rats than Selaginella tamariscina. |
| 语种 | 中文 |
| 公开日期 | 2016-05-03 |
| 源URL | [http://ir.ciac.jl.cn/handle/322003/64485]  |
| 专题 | 长春应用化学研究所_长春应用化学研究所知识产出_学位论文 |
| 推荐引用方式 GB/T 7714 | 徐晨. 治疗高尿酸血症的中药筛选及作用机制研究[D]. 中国科学院长春应用化学研究所. 中国科学院研究生院. 2015. |
入库方式： OAI收割
来源：长春应用化学研究所
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