新疆维药芹菜根抗高脂血症活性部位的制备和体外药效学研究

发布时间：2018-01-06 00:23

本文关键词：新疆维药芹菜根抗高脂血症活性部位的制备和体外药效学研究　出处：《新疆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:优化维药芹菜根活性部位总黄酮提取纯化工艺,对芹菜根提取物进行质量研究,通过体外药效学研究筛选抗HLP活性成分。方法:采用紫外-可见分光光度计法,以总黄酮含量为指标,优化芹菜根活性部位提取纯化工艺;采用理化鉴别方法鉴定活性部位可能含有的化学成分,应用TLC及HPLC定性定量分析芹菜根活性部位化学成分;建立体外L02和HepG2肝细胞脂肪堆积模型筛选芹菜根活性部位抗HLP的活性成分。结果:建立了以芹菜素为参照的紫外-可见分光光度计法,检测波长395nm,回收率99%~104%,确定了芹菜根活性部位提取方法为萃取法;筛选出XDA-1和D101大孔吸附树脂富集纯化芹菜根乙酸乙酯和正丁醇部位中总黄酮;理化鉴别结果表明芹菜根活性部位可能含有有机酸、皂苷、黄酮类及强心苷类化合物;TLC定性研究表明芹菜根乙酸乙酯部位中含有芹菜素、芹菜苷、3’-甲氧基芹菜苷、芹菜素-7-O-β-D-吡喃葡萄糖苷,正丁醇部位中含有芹菜素、芹菜素-7-O-β-D-吡喃葡萄糖苷;HPLC测定3批芹菜根活性部位黄酮成分含量结果表明乙酸乙酯部位含量较高,不同批次芹菜根总黄酮含量具有一定差异性;体外药效学研究结果表明芹菜素、芹菜苷、3’-甲氧基芹菜苷、芹菜素-7-O-β-D-吡喃葡萄糖苷对L02细胞的LD50分别为49.899、541.938、224.359、896.000μg/m L,最佳作用浓度分别为20.919、168.811、51.515、361.00μg/m L,HepG2细胞的LD50分别为64.521、430.226、302.677、961.000μg/m L;最佳作用浓度分别为12.300、139.016、94.607、377.00μg/m L。结论:芹菜根活性部位提取工艺优化及大孔吸附树脂富集纯化工艺研究能够提高总黄酮含量,建立以芹菜素测定总黄酮含量的方法准确度高,重现性好;TLC和HPLC定性定量研究可为芹菜根的质量标准研究提供参考;体外药效学研究结果初步推断芹菜素、芹菜苷、3’-甲氧基芹菜苷、芹菜根乙酸乙酯部位精制物均具明确降低肝细胞TG含量的作用,为后期的调脂机制研究提供实验依据。
[Abstract]:Objective: to optimize the total flavonoids at the active site of celery root extraction and purification process, to study the quality of celery root extract, through in vitro pharmacodynamic studies on screening of anti HLP active ingredient. Method: UV Vis spectrophotometry, the content of total flavonoids as index, optimize the extraction and purification process of the active site of celery root; chemical composition the physical and chemical identification methods for the identification of the active site may contain the chemical composition of the active site of celery root analysis using TLC and HPLC qualitative and quantitative; establishment of L02 and HepG2 of hepatic fat accumulation model of celery root activity screening of anti HLP active ingredient. Results: established with apigenin as UV - visible reference the spectrophotometer method, the detection wavelength of 395nm, the recovery rate of 99%~104%, the method of extracting the active site for the extraction of celery root; screening of enrichment and purification of celery root ethyl XDA-1 and D101 macroporous adsorption resin Total flavonoids of ester and n-butanol; physicochemical identification results showed that the root activity of parts of celery may contain organic acids, saponins, flavonoids and cardiac glycosides; TLC qualitative research shows that celery root of ethyl acetate containing apigenin, Apiin, 3 '- methoxy Apiin apigenin beta -7-O-. -D- glucopyranoside, n-butanol extracts contain apigenin, apigenin -7-O- beta -D- glucopyranoside; HPLC determination of 3 batches of celery root active site of flavonoids was showed higher ethyl acetate content, the content of total flavonoids in celery root in different batches is different; in vitro pharmacodynamic results show that apigenin, Apiin, 3' - methoxy Apiin apigenin, LD50 -7-O- beta -D- glucopyranoside on L02 cells were 49.899541.938224.359896.000 g/m L, the optimal concentration was 20.919168.811,51.515361.00 g/m L HepG2, LD50 cells were 64.521430.226302.677961.000 g/m L; the optimal concentration was 12.300139.016,94.607377.00 g/m L. conclusion: optimization of extraction and macroporous resin purification technology of total flavonoids can improve the root activity parts of celery, method in apigenin content determination of total flavonoids in the high accuracy and good repeatability; study of TLC and HPLC can provide a reference for the qualitative and quantitative study on the quality standard of celery root; in vitro pharmacodynamic results concluded that apigenin, Apiin, 3 '- methoxy Apiin, celery root ethyl acetate extracts have clear parts decreased the TG content in liver cells, and provide experimental basis for research the mechanism of fat.

【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R29

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