中国科学院新疆理化技术研究所机构知识库
Advanced  
XJIPC OpenIR  > 资源化学研究室  > 学位论文
题名: 异常黏液质成熟剂、清除剂的制备及其生物活性研究
作者: 希尔艾力•吐尔逊
答辩日期: 2014-05-30
导师: 阿吉艾克拜尔·艾萨
专业: 有机化学
授予单位: 中国科学院大学
授予地点: 北京
学位: 博士
关键词: 异常黏液质成熟剂 ; 清除剂 ; 制备工艺 ; 成分分析 ; 生物活性
摘要: 维吾尔医学2500多年以来,久存、发展、完善,是因为其具有理论体系即维吾尔医学理论体系。有史以来,维吾尔医学理论对维吾尔医学的发展起了关键的指导作用。目前维吾尔医药事业的发展在国内和我区迈入了一个新的历史性发展时期。今日,维吾尔医药事业的发展是维吾尔医学的迫切需求,深入研究和创新维吾尔医药理论是我们的使命。维吾尔医学认为胆液质(Sapra),血液质(Kan)﹐黏液质(Balgam)﹐黑胆质(Sawda)等四种体液(Hilit)的异常变化导致体内各种物质性病变。其中异常黏液质性病理产物,在血管周围和组织间沉着,产生淤阻,改变组织器官的气质,最终导致中风,老年痴呆症,糖尿病,白癜风,银屑病,单纯性肥胖症,风湿病等难治性疾病。异常黏液质性疾病的治疗原则是根据引起黏液质性疾病的体液分型,首先使用相应的成熟剂(Munziq)使异常黏液质成熟和堆积,然后使用相应的清除剂(Mushil)使成熟的异常黏液质排出体外,使气质(Mizaj)和库外提功能复原,体液平衡,为疾病的治疗奠定基础。本论文主要对异常黏液质成熟剂、清除剂制备工艺、质量标准、化学成分及体内外生物活性进行研究为其进一步的开发和利用提供科学依据。
1. 成熟剂、清除剂制备工艺研究
本论文对成熟剂、清除剂合剂进行了制备工艺研究,以总黄酮、出膏率为指标,采用正交实验法研究了最佳提取工艺。实验结果表明,成熟剂按照传统玫瑰花浸泡提取,参数为加水量20倍,60℃浸泡6h。其余甘草等7味药材回流提取加水量14倍,浸泡时间为24h,与玫瑰花药渣合并,回流提取2次,每次1小时,过滤,合并浓缩,制成1000ml。清除剂制备工艺为按照传统刺糖浸泡,加10倍水,60℃浸泡4小时;阿勃阿 加10倍水,60℃浸泡4小时,滤过,备用;其它药材粉碎成粗粉,加14倍水,浸泡24h后,提取2次,每次1h,合并煎液,滤过,合并上述浸泡液,浓缩,制成1000ml。成熟剂、清除剂合剂pH值控制在4.0-6.0的范围内,采用流通蒸汽灭菌法,100℃灭菌30min,成熟剂合剂的相对密度规定为不得低于1.01(20℃),清除剂合剂的相对密度规定为不得低于1.04(20℃)。成熟剂、清除剂加入防腐剂选用0.20%的山梨酸作为防腐剂。
2.成熟剂、清除剂化学成分HPLC/QTOF-MS/MS及逆流色谱技术分析 采用HPLC/QTOF-MS/MS技术,对成熟剂、清除剂的化学成分初步HPLC分析,根据负离子模式下获得各色谱峰质谱数据,参照文献,对照物质对主要色谱峰进行指认,共分析了成熟剂24个化合物,清除剂21个化合物,其主要化学成分是有机酸类、鞣质类、黄酮及其苷类化合物。对成熟剂、清除剂样品的乙酸乙酯部位进行分离、纯化、鉴定研究,成熟剂乙酸乙酯萃取部位,经过HSCCC一步分离得到纯度比较高的3个化合物并鉴定化合物Ⅰ为没食子酸甲酯,化合物Ⅱ为5,7,4’-三羟基6,8-二异戊烯基异黄酮,化合物Ⅲ为甘草酚;清除剂乙酸乙酯萃取部位,经过HSCCC一步分离得到纯度比较高的3个化合物并鉴定出其中的2个化合物分别为化合物Ⅳ为葫芦素E-2-O-β-D-吡喃葡萄糖苷,化合物Ⅵ为2,5-二甲基-7-羟基色酮。
3.成熟剂、清除剂合剂质量标准的建立
对成熟剂、清除剂合剂进行了质量标准研究。一般检查结果均符合中华人民共和国药典(2010版)合剂项下有关要求。成熟剂对甘草、玫瑰花进行了薄层鉴别,运用HPLC法对没食子酸、鞣花酸、甘草酸铵等3个成分进行含量测定研究;清除剂对阿勃勒、甘草进行了薄层鉴别,采用HPLC法对没食子酸、鞣花酸、甘草酸铵等3个成分进行含量测定研究。
4.成熟剂、清除剂体内外生物活性研究
本文中成熟剂、清除剂及成熟剂醇沉样品进行体外PTP1B、α-葡萄糖苷酶和醛糖还原酶酶体系抑制剂的筛选和ABTS、DPPH自由基清除实验。结果表明这三种样品对PTP1B和α-葡萄糖苷酶有不同程度的抑制作用;而对醛糖还原酶无抑制作用。在清除自由基试验中虽然相对阳性对照维生素C的效果相差较大,但也有一定的清除作用。实验结果表明成熟剂、清除剂、成熟剂醇沉提取物可以作为预防和治疗糖尿病的潜在有效药物。观察成熟剂、清除剂提取物对2型糖尿病大鼠糖、脂代谢的影响,初步探讨了其作用机制。造模成功后按照成熟剂、清除剂维吾尔医临床应用规则,首先灌胃低、中、高三个剂量组的成熟剂提取物4周。结果:与模型组相比,体重及脏器指数:成熟剂中、高剂量组体重升高,高剂量组肝指数降低。糖代谢作用,血糖的变化;成熟剂高剂量组大鼠空腹血糖降低。对血清胰岛素:成熟剂中、高剂量组降低血糖,成熟剂(低、中、高剂量)均降低血清胰岛素,并呈剂量规律性。对胰岛素敏感指数:成熟剂中、高剂量组胰岛素敏感指数升高,且呈剂量规律性。脂代谢作用的相关指标比较:成熟剂低剂量组TG、LDL-C降低,HDL-C升高,成熟剂中、高剂量组TG、TC、LDL-C降低,HDL-C升高,且有剂量规律性。 观察清除剂造模成功后按照维吾尔医临床应用规则首先灌胃成熟剂4周后停止成熟剂,开始灌胃清除剂10天,结果:与模型组相比,体重及脏器指数观察,清除剂组(低、中剂量)体重降低;清除剂高剂量组体重升高、肝指数降低,且有剂量规律性。糖代谢作用,血糖的变化观察,第5周,清除剂高剂量组降低血糖。对血清胰岛素观察,清除剂高剂量组降低血糖,清除剂组(低、中、高剂量)均降低血清胰岛素。对胰岛素敏感指数,清除剂高剂量组胰岛素敏感指数升高。脂代谢作用的相关指标比较,5 周后清除剂低剂量组 TG 、 LDL-C降低, HDL-C升高中、高剂量组 TG 、 HDL-C降低,高剂量 LDL-C升高。病理结果:模型组比较,成熟剂高剂量组及阳性对照组病理变化程度有明显的减轻,范围变小的现象。
英文摘要: For more than 2500 years, the Traditional Uyghur Medicine (TUM)has been able ?to preserve its core value, and continuously develop due to a scientific theoretical ?system of the Uyghur Medicine Theory. Historically, the theory of Uyghur Medicine ?has played a key role in guiding Uyghur Medicine treatment and its development and ?promotion. But after the 19th century, due to various historical and other objective ?reasons, theoretical studies of Uyghur Medicine have been almost stationary. Today, ?development of Uyghur Medicine and the era demands us to deepen the research and ?innovate the theory of Uyghur Medicine in order to accommodate rapid expantion ?of human population and advance in the science and technology.? According to Traditional Uyghur Medicine, abnormal changes of the four body ?liquids such as Sapra(yellow bile), Kan(blood), Balgam(phlegm) and Sawda(black bile) ?causes various substantial lesions in the body. Pathological product of abnormal ?Balgam calming in perivascular tissue, generating silt resistance, changing the ?temperament of the organ tissue ,then eventually leads to refractory diseases such ?as stroke, Alzheimer's disease, diabetes, vitiligo, psoriasis, simple obesity, rheumatism, ?etc. Treatment principles of abnormal Balgam disease is to first use the appropriate ?ripening agent (Munziq) to mature and accumulate abnormal fluid, then to excrete it ?by using the appropriate scavengers (Mushil) and in the end to recover the ?temperament (Mizaj) and the power(Kuwwat), balance body fluid, then lay the ?foundation for the treatment of disease. In this thesis, research was done on the ?preparation process ,chemical composition and quality control of abnormal Balgam ?Munzij and Mushil, and their biological activities in vivo and in vitro to provide the ?scientific basis for their development and utilization.? ?1. Preparation technology of Munziq and Mushil Munziq Preparation technology was as follows: it was prepared by the method of ?water extraction, the best extraction parameters were determined by the extraction ?yield and general flavone, single factor experiments and orthogonal experiments. The ?Rosa rugosa Thunb. follow a traditional method of Uyghur Medicine Soak extract , ?method was water for once. 20 times the water of its own weight , Soak temperature ?was 60 degrees lasted 6 hours. The rest prescription first Souk 24 hours , then reflux ?extraction with water for twice. 14 times the water of its own weight was Each time . ?Each reflux extraction lasted 1 hours. The fried liquid was gathered, then filtered.? Mushil Preparation technology was as follows: it was prepared by the method of ?water extraction, the best extraction parameters were determined by the extraction ?yield and general flavone, single factor experiments and orthogonal experiments. The ?Alhagi pseudoalhagi (M.B.) Desv. And Cassia fistula L. follow a traditional method ?of Uyghur Medicine each Herbs Soak extract , method was water for once. 10 times ?the water of its own weight , Soak temperature was 60 degrees lasted 4 hours. The ?rest prescription first Souk 24 hours , then reflux extraction with water for twice. 14 ?times the water of its own weight was Each time . Each reflux extraction lasted 1 ?hours. The fried liquid was gathered, then filtered. The filtrate was concentrated to a ?relative density of 1.01 Munziq and 1.04 Mushil, and then 2g sorbic acid was added ?and mixed . Finally we adjusted the pH to 4.0 to 6.0 and added water to 1000ml and ?steam sterilization.? ?2. Analysis by LC/QTOF–MS/MS and Chemical Compositions Chemical Compositions of Munziq and Mushil were separated and analysed by ?LC/QTOF–MS/MS in negative electrospray ionisation (ESI) mode at different ?collision energy (CE) values. The maximal structural information was obtained for the ?identi?cation of components. 24 compounds of Munziq and 21 compounds of Mushil, ?including organic acid, tannins and ?avonoids, were identi?ed or partially ?characterised according to accurate mass measurement and reference substance.? We used High Speed Counter-current Chromatography to separate compounds ?from the fraction extracted by ethyl acetate of Munziq and Mushil. Three compounds, ?Methyl gallateⅠ, 5,7,4'-tri-hydroxy-6,8-diprenylf isoflavonesⅡ, Glycyrol were ?isolated from of Munziq. Another two compounds, Cucurbitacins E-2-O-β-D –?Glucopyranosides Ⅳ, 2,5-Dimethyl-7-hydroxychromoneⅥ, were isolated from of ?Mushil. The chemical structures of compounds Ⅰ-Ⅵ were established on the basis of ?a spectroscopic analysis including 1H NMR and13C NMR.? ?3. Research on quality standards of Munziq and Mushil.? We have established quality standards of Munziq and Mushil. Routine inspection ?was in line with the 2010 version of "Chinese Pharmacopoeia" requirement. ?Glycyrrhiza uralensis Fisch., Rosa rugosa Thunb of Munziq, and Glycyrrhiza uralensis ?Fisch., Cassia fistula L.of Mushil were identificated by TLC. In addition, the HPLC ?method of determination the content of gallic acid、ellagic acids、glycyrrhizic acid was ?established.? ?4.Studies on the Biological Activities in vivo and in vitro ? We studied biological activity of Munziq and Mushil in vivo and in vitro. In ?vitro, we investigated the the inhibitory effects on protein tyrosine phosphatase ??1B(PTP1B)、α-glucosidase and human recombinant aldose(HAR) reductase and ?free radical scavenging experiments on ABTS and DPPH. The results showed that ?these samples displayed different inhibitory activity against PTP1B and α?-glucosidase. Howeve,they showed no activity against HAR. They still held great ?promise although the effects on radical scavenging were weak compared with vitamin ?C. The results of present study clearly demonstrated the potential of using for ?prevention and treatment of diabetes.? To observe the effect of Munziq and Mushil extract on glucose and ?lipid metabolism of type 2 diabetes rats and discuss its possible mechanisms .? Animal model of type 2 diabetes was established by a high fat diet with ?intraperitoneal injection of STZ into male SD rats and give low-dose, middle-dose, ?high-dose of 2 Samples extract. The level of Fasting plasma glucose (FPG), Fasting ?insulin (FINS), Total cholesterol (TC), Triglyceride (TG), High density lipoprotein ?cholesterol (HDL-C), Low density lipoprotein cholesterol (LDL-C), Insulin sensitivity ?index (ISI) and the pathological changes of the liver and kidney were evaluated.? Compared with model group, the liver index in Munziq extract group declined, ?the level of FPG declined and ISI increased—except low-dose group of Munziq, And ?dose of regularity. the level of TG,LHL-C of low-dose group of Munziq declined, ?HDL-C was rise . and the levels of TG, TC, LDL-C of all others Munziq extract group ?declined. HDL-C was on rise . Munziq extract group can lighten the pathological ?lesion of the liver and kidney.? Compared with model group, 5 weeks later the liver index in preparation ?process Mushil extract group declined, the level of FPG was decreased and ISI was ?increased in dose dependent manner. The level of TG,LHL-C of low-dose group of ?Mushil declined, HDL-C was increased. and the levels of TG, LDL-C of all others ?Mushil extract group declined, HDL-C was on rise .
内容类型: 学位论文
URI标识: http://ir.xjipc.cas.cn/handle/365002/3453
Appears in Collections:资源化学研究室_学位论文

Files in This Item:
File Name/ File Size Content Type Version Access License
希尔艾力·吐尔逊.pdf(9074KB)学位论文--暂不开放View 联系获取全文

作者单位: 中国科学院新疆理化技术研究所

Recommended Citation:
希尔艾力•吐尔逊. 异常黏液质成熟剂、清除剂的制备及其生物活性研究[D]. 北京. 中国科学院大学. 2014.
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[希尔艾力•吐尔逊]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[希尔艾力•吐尔逊]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
文件名: 希尔艾力·吐尔逊.pdf
格式: Adobe PDF
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Powered by CSpace