学科：中药学,出版时间：2018,导师：龚千锋指导,学位授予单位：江西中医药大学,论文作者：李佟拉嘎著,副题名：,学科专业：,关键词：,馆藏号：,
中文
本文对蒙古族药塔日努(狼毒)的炮制工艺、炮制减毒的原理及饮片质量评价进行了研究，在研究中采用现代分析技术和现代药理学实验，对塔日努生品及其不同炮制品进行了较为全面的化学成分分析、药效学、毒理学等方面的研究，并初步探讨炮制减毒的原理，以期为蒙药塔日努及其不同炮制品的蒙医临床应用及质量评价提供科学依据。 1、蒙药塔日努炮制工艺研究：本论文以蒙药塔日努中狼毒乙素、岩大戟内酯B的含量及共有峰峰面积之和作为优选炮制工艺的考察指标，采用L₉(3⁴)正交试验法，对古文记载的塔日努炮制工艺进行研究，优选炮制过程中辅料用量、炮制温度和加山羊肉的量等因素，结果表明文献记载塔日努牛奶制、酒制、诃子汤炮制方法及工艺稳定、可靠。 2、蒙药塔日努炮制前后化学成分研究：采用飞行时间质谱法对塔日努炮制前后化学成分变化、以及该变化与药效和毒理的相关性进行研究，结果表明塔日努生品及不同炮制品中有35个稳定的共有峰，与生品相比含量降低的化合物分布情况为：奶制品有23个、诃子汤制品有20个、酒制品有19个。这些化学成分对其减毒的机理、快速鉴别生品及其不同炮制品及其质量评价具有一定的理论意义及实用价值。 3、蒙药塔日努炮制前后药效作用研究：采用整体动物实验法，考察塔日努生品及其不同炮制品对正常小鼠小肠推进及泻下作用的影响，判断塔日努生品及其不同炮制品对正常小鼠有无肠推进和“泻下”作用，以及该作用的差异性情况。结果表明在肠推进实验研究方面，塔日努生品及奶，酒制低(0.133g·㎏⁻¹)、高(0.530g·㎏⁻¹)剂量组、诃子汤制高(0.530g·㎏⁻¹)剂量组与空白对照组比较，均有明显促进正常小鼠小肠推进作用，且有极显著性差异(P＜0.01)，诃子汤制低(0.133g·㎏⁻¹)剂量组与空白对照组存在明显的统计差异(P＜0.05)。在泻下指数评价方面，与空白对照组比较奶制品组有显著性差异(P＜0.05)，生品组及酒制品组与空白对照组比较均有极显著性差异(P＜0.01)，诃子汤制品无显著性差异，表明塔日努生品、酒制品及奶制品明显缩短小鼠第一次排便时间，使干粪便总粒数增加，并出现稀粪点。 4、蒙药塔日努炮制前后代谢组学研究：基于HPLC LTQ-Orbitrap分析技术建立代谢指纹轮廓谱方法，采用模式识别技术分析塔日努不同炮制品，对大鼠内源性代谢物差异变化的影响情况和相应的作用机理，同时根据组织形态观察结果探讨炮制前后其毒性表现，确定出相应的靶器官，而进一步讨论其相应的减毒作用机理。根据所得结果评价炮制减毒的效果，为其临床推广应用提供了参考依据。 5、蒙药塔日努炮制前后毒性变化研究：小鼠急性毒性实验结果表明，奶、酒、诃子汤制塔日努较生品塔日努半数致死量大，表明塔日努炮制后毒性降低。大鼠连续灌胃15天1.5425g·㎏⁻¹生品、2.1595 g·㎏⁻¹牛奶、酒、诃子汤制品及3.2393g·㎏⁻¹牛奶，诃子汤制品(高剂量组)后取肝、肾、心、肺、十二指肠，胃、大肠等组织进行病理切片检查，得出奶制品组、奶制品高剂量组、生品组大鼠胃、小肠、大肠、肝、肾、肺、心组织学检查出现一定的病理变化，以胃和肝明显；组别间以生品组更明显；酒制品组、诃子汤制品组、诃子汤高剂量组大鼠各器官损伤不明显。以上结果提示，蒙药塔日努经酒、诃子汤炮制后毒性明显降低，为塔日努临床应用提供了可靠的实验依据。 6、蒙药塔日努生品及炮制品质量评价研究：前后采用HPLC法建立指纹图谱及含量测定分析方法，针对内蒙古地区13批塔日努生品及其不同炮制品的指纹图谱及70％乙醇超声提取物中的狼毒乙素和岩大戟内酯8的含量进行研究，对13批塔日努及其不同炮制品进行了质量评价研究。 关键词：蒙药塔日努；炮制品；化学成分；药效；代谢组学；毒性；质量评价
英文
Ln this thesis, we studied the processing technology, quality evaluation and toxicity-attenuating mechanism of Mongolian Medicine Tarinu. More precisely, we investigated the chemical composition, pharmacodynamics and toxicology studies using the modern analytical technologies and pharmacology approaches to explore the principle of toxicity attenuation effect of Tarinu raw material and its processed products. Finally, we will elucidate the underlying mechanism of toxicity attenuation effect through processing approaches and to provide scientific evidences for clinical application and quality management of Mongolian Medicine Tarinu. 1.Study on processing technology of Mongolian Medicine Tarinu Two main active compounds 2,4-dihydroxy-6-methoxy-3-methyl-acetophenone and jolkinolide B of Tarinu and their sum of common peak area was used to optimize the processing technology, by L₉(3⁴) orthogonal test method. According to the reported ancient processing methods, the factors related to the amount of auxiliary materials, processing temperature and the amount of goat meat were investigated. The results confirm the ancient processing methods that milk, alcohol and Terminalia chebula Retz soup processing methods are stable and reliable 2.Study on the chemical composition analysis of Mongolian Medicine Tarinu before and after processing We used LC-Q-TOF/MS to study the changes of chemical compositions of Mongolian Medicine Tarinu before and after processing and the correlations between the changes to efficacy and Toxicology. The results show that there are 35 stable common peaks in raw Tarinu and different processed products. In contrast to Tarinu raw material, 23 compounds have been reduced in milk processing; 20 compounds have been reduced in Tenninalia chebula Retz soup processing; and 19 compounds have been reduced in alcohol processing. We summarize that the reduction of the chemical components of Tarinu after processing may have significances for the toxicity-attenuating mechanism Tarinu. Also, the findings may assist the rapid identification of raw Tarinu and its prsocesed products and their quality evaluation. Further studies are needed to explore their pharmacological effects and clinical applications. 3.Study on the pharmacodynamics effect of Mongolian Medicine Tarinu before and after Processing This study was aimed to evaluate the raw Tarinu and its processed products on the influence of the small intestine propulsion and diarrhoea in mice. The results show that raw Tarinu, milk processing, alcohol processing could promote the small intestine propulsion with the dose of 0.133g·㎏⁻¹,0.530g·㎏⁻¹ (P＜0.01, P＜0.01, P＜0.01) compared with the blank group. Terminalia chebula Retz soup processing group could enhance the small intestine propulsion (P＜ 0.05) with the dose of 0.133g·㎏⁻¹ raw Tarinu. Milk processing and alcohol processing group could induce the diarrhoea relative to blank group (P＜ 0.05). Tenninalia chebula Retz soup processing group has no effect on inducing diarrhoea (P ＞0.05). 4.Study on the metabolomics of Mongolian Medicine Tarinu before and after processing The metabolic fingerprint profiles were analyzed by HPLC LTQ-Orbitrap analysis technology. The pattern recognition technology was used to analyze the effects of different processed products of Tarinu on the endogenous metabolites in rats and the corresponding mechanisms. At the same time, according to the results of histomorphological observation, the toxicological performances before and after processing were explored. The corresponding target organs were determined, and the corresponding mechanism of attenuation was further discussed. According to the results, the effects of processing and attenuation were evaluated, which provided scientific evidences for their clinical applications. 5.Study on toxicity attenuation effect of Mongolian Medicine Tarinu before and after Processing An acute toxicity test in mice show that processed products of Tarinu has lower toxicity effect than its raw drug. Rats were continuously garaged with 1.5425g·㎏⁻¹ of raw Tarinu, 2.1595g·㎏⁻¹ of milk, alcohol and Terminalia chebula Retz soup processing Tarinu and 3.2393 g·㎏⁻¹ of milk and Terminalia chebula Retz soup processing Tarinu (high dose group) for 15 days. Liver, kidney, heart, lung, duodenum, stomach, large intestine were examined by pathological biopsy. The results show that there are certain pathological changes of the stomach, small intestine, large intestine, liver, kidney, lung, and heart of rats in the raw Tarinu, milk, high dose of milk group, with obvious stomach and liver changes. There is no obvious damage to the organs of rats in the high-dose terminalia chebula Retz soup processing Tarinu group and alcohol processing group. The raw Tarinu group is more obvious among the whole groups. The injury of organs are not obvious in the rats treated with alcohol, terminalia chebula Retz soup processing group and the high-dose processing Tarinu group. The overall results suggest that the toxicity of the Mongolian Medicine Tarinu by alcohol and terminalia chebula Retz soup processing is obviously attenuated, which provides a reliable experimental basis for the clinical application of Tarinu. 6.Study on the quality evaluation of raw Tarinu and its processed products The fingerprinting and content determination methods were established by HPLC. Fingerprinting was studied for 13 batches of Tarinu and its processed products in Inner Mongolia. 2,4-dihydroxy-6-methoxy-3-methyl-acetophenone and jolkinolide B in the 70% ethanol ultrasonic extract and evaluation of the quality of 13 batches of Tarinu and its processed products were carried out preliminarily. Keywords: Mongolian medicine Tarinu ; processed products; chemical component; pharmacodynamic; metabolomics; toxicity;quality evaluation
