人参同工酶的比较研究

【作者】 孙延香；

【导师】 赵雨；

【作者基本信息】 长春中医药大学 ， 中药学， 2010， 硕士

【摘要】 目的：本论文主要通过现代分离纯化技术、分析鉴定技术及多水平活性检测技术对人参中研究较少但活性多样的生物大分子—蛋白及多肽进行生物特性研究,为优化人参的炮制加工工艺,揭示人参的临床物质基础,开发创新药物、保健食品等系列产品提供科学依据及技术支持。方法：采用PAGE电泳胶内酶活染色方法,对园参(鲜参、生晒参)、山参及林下参中氧化还原酶、水解酶、转移酶、裂解酶、异构酶等5种酶系共22种酶进行活性研究。同时选取园参、山参及林下参中活性均较高的7种酶即POD、CAT、SOD、MDH(氧化还原酶类)和EST、ACP、AMY(水解酶类),针对不同产地(10个产地)、不同部位(芦头、主根、须根)及不同年限(4年、5年)的园参(鲜参),进行酶活力测定。结果：四种参中共有13种酶具有清晰酶活性谱带,其中,鲜参11种,生晒参8种,山参9种,林下参7种。过氧化物酶(POD)、过氧化氢酶(CAT)、苹果酸脱氢酶(MDH)、超氧化物歧化酶(SOD)、酯酶(EST)、酸性磷酸酶(ACP)、淀粉酶(AMY)7种酶为四种参共有,乳酸脱氢酶(LDH)、细胞色素氧化酶(CYT)、谷草转氨酶(GOT)3种酶仅为鲜参所有,磷酸葡萄糖变位酶(PGM)和葡萄糖磷酸异构酶(GPI)仅为山参所有,磷酸葡萄糖脱氢酶(PGD)为生晒参和鲜参共有。结论：四种参酶活性的差别,提示人参因栽培方式、炮制方法等的不同,存在一定质量差异,这为人参的品种选择及临床药用起到一定的借鉴作用。不同产地及不同部位人参各种酶活力均有较大差异；不同年限人参各种酶活力虽有一定差异,但差别不大。人参不同产地间的酶活力差异,说明人参因栽培地区不同,存在一定的质量差异,这为人参的规范化种植提供一定科学依据；人参不同部位酶活力的差异,说明各种酶在参体内的分布不同,同时说明芦头、主根及须根均具有不可忽视的作用,这为人参的整体入药及临床筛选奠定了一定的理论基础；4年及5年生人参酶活力差别不大,说明人参采收年限4年和5年均可。更多还原

【Abstract】 Objective:In this study, we conducted experiments on macromolecules of which had varied activities while not being further studied. The biological activity of these macromolecules-namely ginseng proteins and peptides, had been tested via modern separation and purification technology, analysis and identification technology and detection of activity in vitro and in vivo technology in order to provide scientific evidences and technical support for optimizing the process of preparing ginseng, exposing the clinical material base of ginseng and developing a series of ginseng products including innovative drug and health foods etc.Methods:Using in-gel activity staining method of PAGE electrophoresis to study the activity of 22 kinds of isozyme among 5 different enzyme systems consisting of oxidoreductase system, hydrolase system, transferase system, catenase system and isomerase system in wild ginseng, ginseng in forest and cultivated ginseng. Using colorimetric technology of activity detection to test the activity of POD, CAT, SOD, MDH(oxidoreductase system) and EST, ACP, AMY (hydrolase system) which of them appeared to be high in activity in ginseng. Then to make comparison on enzyme activity of ginseng from different sources(10 sources in Jilin province), with different parts (rhizome、taproot and fibrous root) and of different ages (4 years and 5 years), respectively.Results:Results show clear isoenzymes bands of 13 kinds, which participate in 11 kinds of fresh ginseng,8 kinds of dry ginseng,9 kinds of wild ginseng,8 kinds of ginseng in forest. The peroxydase(POD), catalase(CAT), malic dehydrogenase (MDH),Superoxide dismutase (SOD), Esterase (EST), acid phosphatase (ACP) and Amylase (AMY) in all ginsengs; lactate dehydrogenase (LDH), cytochrome oxidase (CYT) and glutamic oxaloacetic transaminase (GOT) only in fresh ginseng; phosphoglucomutase (PGM) and glucose-6-phosphate isomerase (GPI) only in wild ginseng; 6-phosphogluconate dehydrogenase (PGD) in fresh and dry ginseng.Conclusion:Differences in enzyme activity of four kinds of ginseng suggest that there are some quality differences in ginseng because of differences of cultivation methods, processing methods, which play a certain reference of the selection of species of ginseng and clinical application.It turned out that ginseng from different sources and with different parts did show a great difference in enzyme activity among each group. And ginseng of different ages had but not so much difference in enzyme ability. Differences in enzyme activity of ginseng from different origins suggest that there are some quality differences because of cultivation regional difference, which provide certain scientific reference for ginseng standardized planting; Differences of the enzyme activity in ginseng different parts show that various enzymes in the body of the distribution are different,and the role of reed head, main root and fibrous roots can not be ignored, which has laid a theoretical foundation for medicine and clinical screening of the ginsengs; Differences of enzyme activity in 4 and 5 years old ginseng, show that ginseng can be harvested in 4 years or 5years.更多还原