本文關(guān)鍵詞：新疆維藥芹菜根抗高脂血癥活性部位的制備和體外藥效學(xué)研究　出處：《新疆醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 芹菜根 HLP 活性部位 HPLC 體外肝細(xì)胞脂肪堆積模型

【摘要】：目的:優(yōu)化維藥芹菜根活性部位總黃酮提取純化工藝,對(duì)芹菜根提取物進(jìn)行質(zhì)量研究,通過(guò)體外藥效學(xué)研究篩選抗HLP活性成分。方法:采用紫外-可見(jiàn)分光光度計(jì)法,以總黃酮含量為指標(biāo),優(yōu)化芹菜根活性部位提取純化工藝;采用理化鑒別方法鑒定活性部位可能含有的化學(xué)成分,應(yīng)用TLC及HPLC定性定量分析芹菜根活性部位化學(xué)成分;建立體外L02和HepG2肝細(xì)胞脂肪堆積模型篩選芹菜根活性部位抗HLP的活性成分。結(jié)果:建立了以芹菜素為參照的紫外-可見(jiàn)分光光度計(jì)法,檢測(cè)波長(zhǎng)395nm,回收率99%~104%,確定了芹菜根活性部位提取方法為萃取法;篩選出XDA-1和D101大孔吸附樹(shù)脂富集純化芹菜根乙酸乙酯和正丁醇部位中總黃酮;理化鑒別結(jié)果表明芹菜根活性部位可能含有有機(jī)酸、皂苷、黃酮類(lèi)及強(qiáng)心苷類(lèi)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測(cè)定3批芹菜根活性部位黃酮成分含量結(jié)果表明乙酸乙酯部位含量較高,不同批次芹菜根總黃酮含量具有一定差異性;體外藥效學(xué)研究結(jié)果表明芹菜素、芹菜苷、3’-甲氧基芹菜苷、芹菜素-7-O-β-D-吡喃葡萄糖苷對(duì)L02細(xì)胞的LD50分別為49.899、541.938、224.359、896.000μg/m L,最佳作用濃度分別為20.919、168.811、51.515、361.00μg/m L,HepG2細(xì)胞的LD50分別為64.521、430.226、302.677、961.000μg/m L;最佳作用濃度分別為12.300、139.016、94.607、377.00μg/m L。結(jié)論:芹菜根活性部位提取工藝優(yōu)化及大孔吸附樹(shù)脂富集純化工藝研究能夠提高總黃酮含量,建立以芹菜素測(cè)定總黃酮含量的方法準(zhǔn)確度高,重現(xiàn)性好;TLC和HPLC定性定量研究可為芹菜根的質(zhì)量標(biāo)準(zhǔn)研究提供參考;體外藥效學(xué)研究結(jié)果初步推斷芹菜素、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確降低肝細(xì)胞TG含量的作用,為后期的調(diào)脂機(jī)制研究提供實(shí)驗(yàn)依據(jù)。
[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.

【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R29

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