人参蛋白活性研究
摘要
人参(Panax ginseng C. A. Meyer.)是我国传统名贵中草药,具有极高的药用价值和广泛的药理作用。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。
1、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。结果表明,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。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。
2、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。结果表明,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。
3、采用中性缓冲液浸提、硫酸铵分级沉淀、等电点沉淀、SP离子交换柱层析、DEAE离子交换柱层析、G-75凝胶过滤柱层析、高效凝胶过滤色谱(HPGFC)及基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)等方法,对园参(鲜参)中活力较高的SOD进行分离纯化,最终得到分子量为31 kDa的人参SOD二聚体。经改良的邻苯三酚自氧化法测定,其酶活力为9480.43 U/mg,纯化倍数为308.51倍。敏感性试验表明,该酶对H2O2、CHCl3-EtOH、Urea、NaN3等变性剂较敏感;稳定性试验表明,该酶70℃以下具有很好的热稳定性,pH4-9范围内具有很好的pH稳定性;紫外-可见光谱测定,该酶最大吸收峰在278 nm。人参SOD的纯化与性质研究,为人参抗衰老的临床药用研究起到一定的指导作用。
4、采用中性缓冲液浸提、中空纤维膜过滤(截留分子量10 kDa及2 kDa)、pH 5.0缓冲液透析、Sephadex G-75凝胶柱层析等方法,对园参(生晒参)中具有明显促细胞增殖作用的蛋白进行分离纯化,并采用SDS-PAGE、HPGFC、MALDI-TOF-MS等技术对其进行分子量测定及纯度鉴定,最终得到分子量为8000 Da的多肽(人参生长肽)。将该肽作用于小鼠胚胎成纤维细胞(3T3)、人胚肺二倍体细胞(2BS)及人喉癌细胞(Hep-2),并将其细胞活性与其他人参蛋白活性进行比较,结果表明,该肽具有明显的促进3T3细胞增殖作用,提示人参在维持组织结构和参与损伤修复以及预防或治疗心脏疾病等方面具有不可忽视的作用。
5、采用SPF级ICR雄性小鼠进行人参蛋白毒理学检验。通过测定体重及脏器重量等进行不良反应试验,结果表明,人参蛋白对雄性幼年小鼠体重及脏器无明显影响,但具一定的肝脏刺激性,并有促进雄性器官生长的作用;通过经口最大剂量给药,进行急性毒性试验,结果表明,最大给药剂量下各受试小鼠活动正常,无中毒症状,说明人参蛋白安全、无毒。人参蛋白的安全性实验,为人参蛋白的产品开发提供了科学依据,为人参蛋白的功能学研究奠定了实验基础。
6、采用昆明种清洁级小鼠进行人参蛋白功能学检验。
(1)通过二硝基氟苯(DNFB)致小鼠迟发性超敏(DTH)反应及小鼠碳廓清试验,进行人参蛋白增强免疫力功能检验,结果表明,小鼠耳肿胀度及吞噬指数明显增加,说明人参蛋白具有强免疫功能的作用。
(2)通过耐常压缺氧及常温游泳试验,进行人参蛋白提高缺氧耐受力及缓解体力疲劳功能检验,结果表明,小鼠耐缺氧及游泳时间明显延长,说明人参蛋白具有增强耐缺氧及抗疲劳功能的作用。
(3)通过60Co-γ射线辐射小鼠试验,进行人参蛋白对辐射危害的辅助保护功能检验,结果表明,小鼠外周血白细胞数明显增加,骨髓细胞微核数明显减少,说明人参蛋白具有增强抗辐射功能的作用;通过角叉菜胶致小鼠非特异性急性炎症反应试验,进行人参蛋白缓解炎症功能检验,结果表明,小鼠足趾肿胀度无明显降低,说明人参蛋白不具有明显的抗炎作用。
(4)通过醋酸致小鼠扭体试验及热板致小鼠舔足试验,进行人参蛋白缓解疼痛功能检验,结果表明,小鼠出现扭体时间明显延长,舔足次数明显减少,说明人参蛋白具有一定的镇痛作用。
(5)通过测定辐射小鼠及正常小鼠灌服人参蛋白后SOD活性及丙二醛(MDA)含量,进行人参蛋白抗氧化功能检验,结果表明,SOD活性升高、丙二醛(MDA)含量降低,说明人参蛋白具有提高抗氧化功能的作用。
综上所述,本论文主要针对人参中的大分子成分—蛋白质进行研究。通过对不同来源人参进行同工酶活力比较,为人参的种质资源鉴定提供酶学依据;通过对人参蛋白进行毒理学与功能学研究,为人参蛋白类药物的开发及应用提供药理学依据。另外,本论文首次比较系统的从人参中分离得到SOD及生长肽纯品,并分别进行理化性质与活性研究,为人参中具有生物活性蛋白的纯化与鉴定奠定了理论与实验基础。
Ginseng(Panax ginseng C. A. Meyer.) is a traditional Chinese herbal medicines and it has high medicinal value and wide range of pharmacological effects. 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.
1、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. 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. 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.
2、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. 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.
3、Using method of neutral buffer extraction and ammonium sulfate fractionation to extract crude enzyme from ginseng and then using isoelectric point precipitation, SP ion-exchange chromatography, DEAE ion-exchange chromatography, G-75 gel filtration column chromatography, high performance gel filtration chromatography(HPGFC) and matrix-asisted laser desorption ionization/time of flight mass spectrometry(MALDI-TOF-MS) to further isolate and purify to get a homodimer of SOD with molecule weight of 31 kDa. The activity of the enzyme tested by improved Pyrogallol autoxidation method appeared to be 9480.43 U/mg, inferring that the purification factor was 308.51. The enzyme was sensitive to detergent as H2O2、CHCl3-EtOH、Urea、NaN3 etc and exhibited high thermo stability (70℃) over the pH range from 4.0 to 9.0. Its maximum absorption wavelength was 278 nm and it was sensitive to hydrogen peroxide. The studies of purification and characterization of ginseng, play a guiding role for clinical medicinal research of ginseng antiaging.
4、Ginseng protein which was extracted by neutral pH buffer and hollow fiber membrane filtration method had been isolated and purified via acidic buffer(pH 5.0) dialysis、hollow fiber membrane ultrafiltration(10 kDa and 2 kDa) and sephadex G-75 gel-filtration chromatography. Then conducting the determination of molecule weight and identification of purification on SDS-PAGE、HPGFC and MALDI-TOF-MS, resulting in a polypeptide(ginseng growth peptide) with molecule weight of 8000Da. Comparing the functional activity on mouse embryonic fibroblasts cells (3T3)、human embryo lung diploid cells (2BS) and human laryngeal carcinoma cells (Hep-2) of this polypeptide with other ginseng proteins (proteinⅠ,Ⅱ, andⅢ), leading to the result that this polypeptide did best in promoting the growth of normal cells. The purification of ginseng growth peptide and significantly promoting the proliferation of 3T3 cells, suggest that the role of ginseng in maintaining the organizational structure and participation in damage repair and prevent or treat heart disease can not be ignored.
5、To conduct toxicology tests of ginseng protein on ICR male mouse of SPF level. The results of adverse reaction test suggested that though having certain stimulating effects on liver, ginseng protein either had no obvious influences on the weight and visceral organs of juvenile male mice or promote the growth of Male genitals. Furthermore acute toxicity test showed that mice of maximum dose acted like normal mice without poisoning, indicating that ginseng protein was safe and non-toxicity. The safety experiments of ginseng protein provided a scientific basis for product development of ginseng protein, and layed an experimental foundation of the functional studies of ginseng protein.
6、To conduct function tests of ginseng protein on Kunming mice of cleaning level. The results of dinitrofluorobenzene (DNFB) induced delayed-type hypersensitivity (DTH) in mice and mice carbon clearance experiment suggested that the ear swelling degree and phagocytic index had largely increased, informing of the immune enhancing function of ginseng protein; Swimming test at room temperature and hypoxia tolerance test showed that ginseng protein had an ability in enhancing hypoxia function and alleviating physical fatigue because of the prolongation of swimming and hypoxia tolerant time. The radiation of 60Co-γray on mice greatly increased the number of peripheral blood leucocyte and sharply decreased the number of bone marrow micronucleus in mice, informing of anti-radiation function of ginseng. The experiment of carrageenan-induced non-specific acute inflammation in mice showed decreasing trend in mouse toe swelling degree with no statistical significance, indicating that ginseng protein had no obvious anti-inflammatory effects. Acetic acid-induced writhing test and pain threshold measuring test by hot-plate in mice showed that the writhing time of mice had been greatly prolonged and the number of paw-licking had decreased, indicating that ginseng protein had certain effect on easing pain. In addition, our studies also found that after being fed with ginseng protein, normal mice and the radiated ones showed higher SOD activity on the contrary to lower MDA concentration, proving that ginseng protein had the ability in promoting antioxidant function.
In summary, The research focuses on bioactive macromolecules of ginseng protein. According to compare with the activity of isozyme from different sources of ginsengs, provides enzymology basis for germplasm resource identification of ginseng. According to function and toxicology testing of ginseng protein, provide a pharmacological basis for the development and applications of ginseng protein drugs. In addition, this research systematically isolated and purified SOD and growth peptide from ginseng, and carried out physical and chemical properties and active research respectively, which layed a theoretical and experimental foundation for purification and identification of the biological activity protein of ginseng.
1、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。结果表明,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。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。
2、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。结果表明,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。
3、采用中性缓冲液浸提、硫酸铵分级沉淀、等电点沉淀、SP离子交换柱层析、DEAE离子交换柱层析、G-75凝胶过滤柱层析、高效凝胶过滤色谱(HPGFC)及基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)等方法,对园参(鲜参)中活力较高的SOD进行分离纯化,最终得到分子量为31 kDa的人参SOD二聚体。经改良的邻苯三酚自氧化法测定,其酶活力为9480.43 U/mg,纯化倍数为308.51倍。敏感性试验表明,该酶对H2O2、CHCl3-EtOH、Urea、NaN3等变性剂较敏感;稳定性试验表明,该酶70℃以下具有很好的热稳定性,pH4-9范围内具有很好的pH稳定性;紫外-可见光谱测定,该酶最大吸收峰在278 nm。人参SOD的纯化与性质研究,为人参抗衰老的临床药用研究起到一定的指导作用。
4、采用中性缓冲液浸提、中空纤维膜过滤(截留分子量10 kDa及2 kDa)、pH 5.0缓冲液透析、Sephadex G-75凝胶柱层析等方法,对园参(生晒参)中具有明显促细胞增殖作用的蛋白进行分离纯化,并采用SDS-PAGE、HPGFC、MALDI-TOF-MS等技术对其进行分子量测定及纯度鉴定,最终得到分子量为8000 Da的多肽(人参生长肽)。将该肽作用于小鼠胚胎成纤维细胞(3T3)、人胚肺二倍体细胞(2BS)及人喉癌细胞(Hep-2),并将其细胞活性与其他人参蛋白活性进行比较,结果表明,该肽具有明显的促进3T3细胞增殖作用,提示人参在维持组织结构和参与损伤修复以及预防或治疗心脏疾病等方面具有不可忽视的作用。
5、采用SPF级ICR雄性小鼠进行人参蛋白毒理学检验。通过测定体重及脏器重量等进行不良反应试验,结果表明,人参蛋白对雄性幼年小鼠体重及脏器无明显影响,但具一定的肝脏刺激性,并有促进雄性器官生长的作用;通过经口最大剂量给药,进行急性毒性试验,结果表明,最大给药剂量下各受试小鼠活动正常,无中毒症状,说明人参蛋白安全、无毒。人参蛋白的安全性实验,为人参蛋白的产品开发提供了科学依据,为人参蛋白的功能学研究奠定了实验基础。
6、采用昆明种清洁级小鼠进行人参蛋白功能学检验。
(1)通过二硝基氟苯(DNFB)致小鼠迟发性超敏(DTH)反应及小鼠碳廓清试验,进行人参蛋白增强免疫力功能检验,结果表明,小鼠耳肿胀度及吞噬指数明显增加,说明人参蛋白具有强免疫功能的作用。
(2)通过耐常压缺氧及常温游泳试验,进行人参蛋白提高缺氧耐受力及缓解体力疲劳功能检验,结果表明,小鼠耐缺氧及游泳时间明显延长,说明人参蛋白具有增强耐缺氧及抗疲劳功能的作用。
(3)通过60Co-γ射线辐射小鼠试验,进行人参蛋白对辐射危害的辅助保护功能检验,结果表明,小鼠外周血白细胞数明显增加,骨髓细胞微核数明显减少,说明人参蛋白具有增强抗辐射功能的作用;通过角叉菜胶致小鼠非特异性急性炎症反应试验,进行人参蛋白缓解炎症功能检验,结果表明,小鼠足趾肿胀度无明显降低,说明人参蛋白不具有明显的抗炎作用。
(4)通过醋酸致小鼠扭体试验及热板致小鼠舔足试验,进行人参蛋白缓解疼痛功能检验,结果表明,小鼠出现扭体时间明显延长,舔足次数明显减少,说明人参蛋白具有一定的镇痛作用。
(5)通过测定辐射小鼠及正常小鼠灌服人参蛋白后SOD活性及丙二醛(MDA)含量,进行人参蛋白抗氧化功能检验,结果表明,SOD活性升高、丙二醛(MDA)含量降低,说明人参蛋白具有提高抗氧化功能的作用。
综上所述,本论文主要针对人参中的大分子成分—蛋白质进行研究。通过对不同来源人参进行同工酶活力比较,为人参的种质资源鉴定提供酶学依据;通过对人参蛋白进行毒理学与功能学研究,为人参蛋白类药物的开发及应用提供药理学依据。另外,本论文首次比较系统的从人参中分离得到SOD及生长肽纯品,并分别进行理化性质与活性研究,为人参中具有生物活性蛋白的纯化与鉴定奠定了理论与实验基础。
Ginseng(Panax ginseng C. A. Meyer.) is a traditional Chinese herbal medicines and it has high medicinal value and wide range of pharmacological effects. 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.
1、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. 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. 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.
2、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. 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.
3、Using method of neutral buffer extraction and ammonium sulfate fractionation to extract crude enzyme from ginseng and then using isoelectric point precipitation, SP ion-exchange chromatography, DEAE ion-exchange chromatography, G-75 gel filtration column chromatography, high performance gel filtration chromatography(HPGFC) and matrix-asisted laser desorption ionization/time of flight mass spectrometry(MALDI-TOF-MS) to further isolate and purify to get a homodimer of SOD with molecule weight of 31 kDa. The activity of the enzyme tested by improved Pyrogallol autoxidation method appeared to be 9480.43 U/mg, inferring that the purification factor was 308.51. The enzyme was sensitive to detergent as H2O2、CHCl3-EtOH、Urea、NaN3 etc and exhibited high thermo stability (70℃) over the pH range from 4.0 to 9.0. Its maximum absorption wavelength was 278 nm and it was sensitive to hydrogen peroxide. The studies of purification and characterization of ginseng, play a guiding role for clinical medicinal research of ginseng antiaging.
4、Ginseng protein which was extracted by neutral pH buffer and hollow fiber membrane filtration method had been isolated and purified via acidic buffer(pH 5.0) dialysis、hollow fiber membrane ultrafiltration(10 kDa and 2 kDa) and sephadex G-75 gel-filtration chromatography. Then conducting the determination of molecule weight and identification of purification on SDS-PAGE、HPGFC and MALDI-TOF-MS, resulting in a polypeptide(ginseng growth peptide) with molecule weight of 8000Da. Comparing the functional activity on mouse embryonic fibroblasts cells (3T3)、human embryo lung diploid cells (2BS) and human laryngeal carcinoma cells (Hep-2) of this polypeptide with other ginseng proteins (proteinⅠ,Ⅱ, andⅢ), leading to the result that this polypeptide did best in promoting the growth of normal cells. The purification of ginseng growth peptide and significantly promoting the proliferation of 3T3 cells, suggest that the role of ginseng in maintaining the organizational structure and participation in damage repair and prevent or treat heart disease can not be ignored.
5、To conduct toxicology tests of ginseng protein on ICR male mouse of SPF level. The results of adverse reaction test suggested that though having certain stimulating effects on liver, ginseng protein either had no obvious influences on the weight and visceral organs of juvenile male mice or promote the growth of Male genitals. Furthermore acute toxicity test showed that mice of maximum dose acted like normal mice without poisoning, indicating that ginseng protein was safe and non-toxicity. The safety experiments of ginseng protein provided a scientific basis for product development of ginseng protein, and layed an experimental foundation of the functional studies of ginseng protein.
6、To conduct function tests of ginseng protein on Kunming mice of cleaning level. The results of dinitrofluorobenzene (DNFB) induced delayed-type hypersensitivity (DTH) in mice and mice carbon clearance experiment suggested that the ear swelling degree and phagocytic index had largely increased, informing of the immune enhancing function of ginseng protein; Swimming test at room temperature and hypoxia tolerance test showed that ginseng protein had an ability in enhancing hypoxia function and alleviating physical fatigue because of the prolongation of swimming and hypoxia tolerant time. The radiation of 60Co-γray on mice greatly increased the number of peripheral blood leucocyte and sharply decreased the number of bone marrow micronucleus in mice, informing of anti-radiation function of ginseng. The experiment of carrageenan-induced non-specific acute inflammation in mice showed decreasing trend in mouse toe swelling degree with no statistical significance, indicating that ginseng protein had no obvious anti-inflammatory effects. Acetic acid-induced writhing test and pain threshold measuring test by hot-plate in mice showed that the writhing time of mice had been greatly prolonged and the number of paw-licking had decreased, indicating that ginseng protein had certain effect on easing pain. In addition, our studies also found that after being fed with ginseng protein, normal mice and the radiated ones showed higher SOD activity on the contrary to lower MDA concentration, proving that ginseng protein had the ability in promoting antioxidant function.
In summary, The research focuses on bioactive macromolecules of ginseng protein. According to compare with the activity of isozyme from different sources of ginsengs, provides enzymology basis for germplasm resource identification of ginseng. According to function and toxicology testing of ginseng protein, provide a pharmacological basis for the development and applications of ginseng protein drugs. In addition, this research systematically isolated and purified SOD and growth peptide from ginseng, and carried out physical and chemical properties and active research respectively, which layed a theoretical and experimental foundation for purification and identification of the biological activity protein of ginseng.
引文
[1]石威.不同生长期人参中化学成分及农药残留的研究[D].吉林大学,2007.
[2]周小红.人参只卖萝卜价,东北参为何难敌高丽参[J].农村百事通,2007(6):6-8.
[3] Liu M, Zhang J. Effects of ginsenoside Rb1 and Rg1 on synaptosomal free calcium level, ATPase and calmodulin in rat hippocampus [J]. Chin Med J (Engl), 1995,108(7): 544-547.
[4]吴柏林,徐祝伟.人参及其复方制剂的抗衰老研究进展[J].时珍国医国药, 1997, 8(6): 560.
[5]程俊霖,周黎明,朱玲,等.人参茎叶总皂苷对衰老小鼠的作用研究[J].四川生理科学杂志,2004,26(3):97-99.
[6]崔巍,赵洪艳,王燕嬉.人参皂甙抗衰老的研究进展[J].中国老年学杂志,2006,26(11):1578-1581.
[7]徐承水.人参抗衰老作用的实验研究[J].赣南师范学院学报,2000(3):54-56.
[8]陈冠敏,陈小萍,何聆,等.人参片对大鼠血脂水平及延缓衰老作用的实验研究[J].实验预防医学,2002,9(4):311-313.
[9]王红丽,吴铁,吴志华,等.人参皂甙抗皮肤衰老作用实验研究[J].广东药学院学报,2003,19(1):25-27.
[10]黄宗锈,陈冠敏,林蔚.人参皂甙延缓衰老的实验研究[J].实用预防医学,2001,8(5):385.
[11]潘鑫鑫,郑卫红,傅万玉,等.人参总皂甙抗衰老作用的实验研究[J].中国老年学杂志,1995,15(5):288-290.
[12]李凤人.人参挥发油对肿瘤的作用[C].国际人参研讨会论文集,长春,1992,179.
[13]高峰.人参与人参皂苷Rh2-恶性肿瘤治疗的新观点[J].人参研究,2001,13(4): 17-18.
[14]陶丽华,高峰,刘红岩,等.20(R)-人参皂苷Rh2抗癌细胞转移实验研究[J].人参研究,2002,14(4):17-18.
[15]张仲苗,江波,郑筱祥.人参皂苷Rg3对肿瘤放疗患者外周血淋巴细胞的体外免疫增强作用[J].中国药学杂志,2004,39(4):261-264.
[16] Huang D. Saponins and sapogenins for use in combination therapy for cancer [P]. U S,WO/2004/056379, 2004-07-08.
[17]刘忞.人参皂甙Rg1抗衰老和促智作用及其机制研究[J].生理科学进展, 1996, 27(2): 139-142.
[18]刘单,张均田.人参皂苷Rg_1对老年大鼠行为活动的改善作用[J].中国药学杂志, 1996,31(8):464-467.
[19]胡圣望,胡勇,胡王平.人参皂甙Rg1对慢性应激大鼠空间学习记忆能力的影响[J].四川中医,2004,22(3):14-15,16.
[20]沈仲理.人参对人体增强免疫力和抗病能力的研究[J].上海预防医学杂志, 1995, 7 (12):529-530.
[21]吴浩,林洪生,裴迎霞,等.人参皂甙Rg3对荷瘤及环磷酰胺化疗小鼠粘膜免疫力影响[J].中国肿瘤,2006,15(6):369-371.
[22]武淑芳,睢大员,于晓风,等.西洋参叶20s-原人参二醇组皂苷抗实验性心肌缺血作用及其机制[J].中国药学杂志,2002,37(2):100-103.
[23]田建明,李浩,叶金梅,等.人参皂苷Rg2对大鼠化学性心肌缺血的影响[J].中国中药杂志,2003,28(12):1191-1192.
[24]张秋梅,张喆,于德民,等.人参糖肽治疗2型糖尿病的临床观察[J].中国现代医学杂志, 2003,13(6):59,62.
[25] Kim D H, Bae E A, Han M J, et al. Novel use of the extract of processed ginseng and saponin isolated therefrom [P]. U S, 20050232908, 2005-10-20.
[26]金慧,周经纬,娄子恒.人参产业与区域经济发展[J].人参研究,2004,16(1):2-6.
[27]高涛,王立莹.我国人参市场现状及前景展望[J].人参研究,2006, (3):5-6.
[28]李向高.人参产业的创新战略[J].吉林农业大学学报,2002,24(5):1-5.
[29]张中朋,刘张林.人参出口额快速增长西洋参进口额高于出口—2008年1~6月份人参、西洋参进出口情况分析[J].中国现代中药,2008,10(8):35,44.
[30]戴昀弟.人参产业可持续发展对策探讨[J].社会科学战线·东北经济研究, 2007(5): 68- 70.
[31]孙峥,周滋养.吉林省人参产业发展探析[J].安徽农业科学,2008,36(23):10019-10021
[32]于志斌,罗扬.人参国际市场分析与出口形势预测[J].中国现代中药,2009,11(7):45,50.
[33]曹广成.中韩人参价值对比研究[J].中国科技博览,2008(24):251-253.
[34]张中朋,刘张林.2007年我国人参出口形势分析及展望[J].中国现代中药, 2008, 10(3): 44-45.
[35]屈文平.吉林省人参生产与出口情况的调研报告[J].对外经贸统计,2005(5):38-41.
[36]李刚,蔡天智.浅谈人参出口问题[J].中药研究与信息,2003,5(8):59-61.
[37]那中凯.国际市场上人参商战策略初探[J].中国医药情报,1999,5(3):166-170.
[38]杨朝丹,郭庆海.吉林省人参加工业发展探析[J].沈阳农业大学学报:社会科学版, 2007, 9(1):16-19.
[39]杨朝丹,丛媛媛.吉林省人参加工企业发展现状、问题及对策研究[J].商情, 2009 (24):33,71.
[40]赵文洋,孙琛.吉林省人参龙头企业发展策略探析[J].现代商业,2009(30):56-57.
[41]仲伟同,迟美丽.人参产业发展的根本出路在于药食同源[J].人参研究,2009(1):26-27.
[42]金玉姬.延边人参加工业存在的问题及对策[J].农村经济与科技,2008,19(12):65,68
[43] Barnes T, Goodacre J, Moots R J, et a1. Therapeutic targeting of cytokines [J].Cytokine, 2006,35(3/4):217-219.
[44]蒋建平.我国蛋白质的综合开发利用[J].中国食物与营养,1995(1):9-12
[45]马静,葛熙,昌增益.蛋白质功能研究:历史、现状和将来[J].生命科学,2007,19(3):294-300.
[46]查锡良.医学分子生物学[M].北京:人民卫生出版社,2003:245-299.
[47]催凌飞,王遂.蛋白质及其水解物的分析应用[J].哈尔滨商业大学学报,2002, 18 (1): 117-120.
[48] Cavazzana-Calvo M, Thrasher A, Mavilio F. The future of gene therapy: Balancing the risks and benefits of clinical trials [J]. Nat, 2004, 427:779-781.
[49] Mathe C, Sagot M F, Schiex T, eta1. Current methods of gene prediction, their strengths and weaknesses[J]. Nucleic Acids Res, 2002, 30(19):4103-4117.
[50] Blattner F R, Plunkett III G, Bloch C A, et a1. The complete genome sequence of Escherichia coli K-12 [J]. Science, 1997, 277(5331):1453-1474.
[51] Wood V, Gwilliam R, Rajandream M A, eta1. Thegenome sequence of Schizosaccharo myces pombe[J]. Nature, 2002, 415(6874):871-880.
[52] Waterstone R. Genome sequence of the nematode C. elegans: a platform for investingating biology[J]. Science, 1998, 282 (5396): 2012- 2018.
[53] Adams M D, Celniker S E, Holt R A, et a1. The genome sequence of Drosophila melanogaster[J]. Science, 2000, 287:2185-2195.
[54] Arabidopsis Genome Initiative. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana[J]. Nature, 2001,408(6814):796-815.
[55] Venter J C, Adams M D, Myers E W, et al. The sequence of the human genome[J]. Science, 2001, 291(5507):1304-1351.
[56]阚欢,李贤忠,陆斌.辣木叶蛋白质提取工艺研究[J].西部林业科学,2007,36(1):106-107.
[57]张晓辉,章克昌.非水溶性茶蛋白的提取工艺研究[J].食品研究与开发,2005,26 (3):64—67.
[58]刘香萍,李国良,王凤军.酸热法提取苜蓿叶蛋白初探[J].实验研究, 2007(1):10-11.
[59]刘高强,周虎,魏美才.稀盐法提取马尾松毛虫蛹中的蛋白质[J].食品科技, 2007, (9): 132-134.
[60]刘朋,邱芳萍.梅花鹿胎衣中蛋白质的提取分离[J].现代食品科技,2006,22(4):153-155.
[61]左玉萍,冯成利,党蕊叶.猕猴桃蛋白酶提取的工艺技术研究[J].西北农业学报, 2006, 15(6):127-129.
[62]张巍,李红艳,马晶,等.人参水溶性蛋白的纯化工艺研究[J].吉林农业大学学报, 2008, 30(1):36-39.
[63]李红艳,赵雨,张巍,等.人参水溶性蛋白对几种细胞增殖的影响[J].吉林农业大学学报,2008,30(5):705-707,711.
[64]唐任能,赵雨,孙晓迪,等.梅花鹿鹿茸、鹿托盘及鹿骨水溶性总蛋白比较研究[J].吉林中医药,2008,28(4):295-296.
[65]幺宝金.人参蛋白的提取分离及性质研究[D].长春:长春中医药大学,2009.
[66]孟林,田景振.动物组织蛋白质提纯方法的研究进展[J].食品与药品,2009,11(5):53-55.
[67] Damerval C, Vienne D D, Zivy M, et a1. Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins [J]. Electrophoresis,1986,7(1):52-54.
[68] Wu F S, Wang M Y. Extraction of proteins for sodium dodecyl sulfate-polyacrylamide gel electrophoresis from protease-rich plant tissues[J]. Anal Biochem. 1984(139):100-103.
[69] Nandaknmar M P, Shen J, Raman B, et a1. Solubilization of trichloroacetic acid (TCA) precipitated microbial proteins via naOH for two-dimensional electrophoresis[J]. J Proteome Res, 2003,2(1):89-93.
[70] Lee S J, Oh P S, Ko J H, et a1. Glycoprotein isolated from Gardenia jasminoides Ellis has a scavenging activity against oxygen radicals and inhibits the oxygen radical-linduced protein kinase C alpha and nuclear factor-kappa B in NIH/3T3 cells[J]. Environ Toxicolo Phar, 2006, 21(1):8-21.
[71]胡兴昌,唐仕华.板蓝根生药中蛋白质提取方法的比较[J].上海师范大学学报, 2004 (12):66-68.
[72]郭雪松,黄小杰,王烁,等.醇法大豆浓缩蛋白提取工艺的优化[J].食品工业科技, 2007, 28 (5):181-182.
[73] G?rg A, Weiss W, Dunn M J. Current two-dimensional electrophoresis technology for proteomics [J]. Proteomics, 2004, 4(12):3665-3685.
[74]王川,李燕,马志英,等.几种酶法从猪皮中提取胶原蛋白的对比研究[J].食品科学, 2007,28(1):201-204.
[75]沈莲清,黄光荣,王向阳,等.茶渣中蛋白质酶法提取工艺[J].食品与生物技术学报, 2006,25 (6):7-12.
[76]陈凤莲,方桂珍.不同方法分离小麦麸皮中蛋白质的研究[J].粮食加工, 2009, 34(4): 28-30,35.
[77]李二凤,何小维,罗志刚.胶原蛋白的提取工艺研究[J].食品研究与开发,2006, 27 (3): 64-65,63.
[78] Moulton K J, Wang L C. A pilot plant study ofcontinuous ultrasonic extraction of soybean protein [J]. J Food Sci, 1982,47(4):1127-1129.
[79]刘婷婷,王海灵,张艳荣.玉米超氧化物歧化酶超声波提取技术的研究[J].食品科学, 2007,28(8):156-159.
[80]龚丽芬,龚文贤.双水相分离提取胃蛋白酶和琼脂糖的初步研究[J].化学与生物工程,2006,23(11):36-38.
[81]赵晓燕,陈复生,薛文通.不同反胶束体系提取大豆蛋白质的研究[J].食品科学, 2007,28(4):109-111,112.
[82]李建武,萧能愿,余瑞元,等.生物化学实验原理和方法[M].北京:北京大学出版社, 1994: 160-176.
[83]郭颖娜,孙卫.蛋白质含量测定方法的比较[J].河北化工,2008,31(4):36-37.
[84]杨玉芳.蛋白质含量测定方法[J].明胶科学与技术,2007,27(2):99.
[85]薛志欣,杨桂朋,马晓梅,等.分光光度法测定藻胆蛋白含量的研究[J].鲁东大学学报(自然科学版),2008,24(3):250-253.
[86]康东周,张善玉,全仙花.Lowry法测定官颈癌片中蛋白质的含量[J].现代医药卫生, 2005,21(23):3210-3211.
[87]陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2002:166-167.
[88]曹艳,黄必胜.半夏总蛋白含量的紫外吸收法测定[J].湖北中医杂志,2005,27(7):48.
[89]王淡兮,孙秀兰.蛋白质定量检测方法的探讨[J].粮食与食品工业,2009,14(4):49-51.
[90] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantifies of protein utilizing the principle of protein-dye binding [J]. Anal Biochem, 1976,7(72):248-254.
[91]曲春香,沈颂东,王雪蜂,等.用考马斯亮蓝测定植物粗提液中可溶性蛋白质含量方法的研究[J].苏州大学学报(自然科学版),2006,22(2):82.
[92]汪良驹,姜卫兵,祝海阔,等.银杏叶片可溶性蛋白质测定非直线性原因分析[J].果树学报,2002,19(1):32.
[93]刘雅超,刘旭,白鸿源,等.蛋白质定量分析的研究进展[J].中国民族民间医药, 2009 (8): 49-50.
[94]王超英,周如真,戴国华,等.胸腺肽注射液分子量测定[J].中国生化药物杂志, 2000, 21(1):33-34.
[95] Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis[J]. J Biol Chem, 1969, 244 (16):4406-4412.
[96] Swank R T, Munkres K D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem, 1971, 39(2):462-477.
[97]崔韶晖,王艳颖,崔杰,等.大连湾牡蛎蛋白的分离提取及分子量测定[J].大连民族学院学报,2005,7(1):53-54,73.
[98]陈振江,殷丹,曹艳,等.厚朴种子蛋白质成分分子量测定[J].中成药, 2005, 27(6): 711- 712.
[99]陈绍农,潘远江,陈耀祖.多肽及蛋白质质谱分析新进展[J].质谱学报,1995, 16 (3): 15-21.
[100]Werner E, Bo U R, Sundqvist P, et al. Directional correlation between the primary particle and ejected molecular ions in electronic sputtering of large organic molecules [J]. Phys Rev B, 1989, 39(1):763-766.
[101]解建勋,蒲小平,李玉珍,等.蛋白质组分析技术进展[J].生物物理学报,2001,17:19 -26.
[102]Zhao S K, Somayajula K V, Sharkey A G, et al. Novel method for matrix-assisted laser mass spectrometry of proteins [J]. Anal Chem,1991,63(5):450-453.
[103]赵善楷,钟峰,张晓红.基体辅助激光解析质谱法(MALDI-MS)在生物化学中的应用[J].质谱学报,1996,17(2):1-7.
[104]刘慧敏,赖志辉,黎军,等.碎片结构分析在MALDI TOF MS法测定多肽序列中的应用[J].生物化学与生物物理学报, 2000 (32):179-182.
[105]郭元平,王书臻,曹振岭,等.榆叶梅果仁蛋白质的提取研究[J].农业与技术,2006, 26 (2) :88-89.
[106]0' Farrell P. High resolution two-dimensional electrophoresis of proteins[J]. J Biol Chem, 1975,250(10):4007-4021.
[107]Cordwell S J, Nouwens A S, Verrills N M, et al. Subproteomics based upon protein cellular location and relative solubilities in conjunction with composite two-dimensional electrophoresis gels[J]. Electrophoresis,2000, 21(6):1094-1103.
[108]Wang W, Scali M, Vignani R, et a1. Protein extraction for two-dimensional electrophoresis from olive leaf, a plant tissue containing high levels of interfering compounds [J]. Electrophoresis,2003,24(14):2369-2375.
[109]Zhao C F, Wang J Q, Cao M L, et a1. changes in rice leaves during development of field-grown rice plants[J]. Proteomics, 2005,5(4):961-972.
[110]安娜,赖钟雄,郭志雄.龙眼体细胞胚胎发生过程中特异表达蛋白的双向电泳分析[J].福建农林大学学报(自然科学版),2007,36(3):244-249.
[111]乐寅婷,李梅,陈倩,等.油菜叶片蛋白质组对机械损伤应答的初步分析[J].植物生态学报,2008,32(1):220-225.
[112]周蕴薇,董文珂,刘艳萍,等.山茶叶片可溶性蛋白双向电泳技术的建立[J].生物技术通报,2008(2):127-132.
[113]范海延,陈捷,张春宇,等.适于黄瓜叶片蛋白质组分析的双向电泳方法最佳条件的研究[J].沈阳农业大学学报,2008,39(3):365-367.
[114]罗泽宇,杨粤军,刘选明.拟南芥蛋白质组研究中双向电泳技术条件的优化[J].激光生物学报,2008,l7(4):539-543.
[115]古卓良,周国华,张晓丹,等.缓冲液种类对毛细管区带电泳分离蛋白质的影响[J].解放军药学学报,2003,19(6):404-407.
[116]朱京妃,夏明.毛细管电泳技术在多肽和蛋白质检测中的应用[J].农产品加工?创新版, 2009(6):54-56.
[117]Colao A, Somma C D, Salerno M, et a1. The Cardiovascular Risk of GH-Deficient Adolescents [J]. J Clin Endocrinol Metab, 2002, 87(8): 3650- 3655.
[118]孙晓东,房泽海,任悦欣.蛋白多肽类药物给药途径及剂型的研究进展①[ J].中外医疗,2009,21(7):160-162.
[119]金城,王海涛,肖小河.蛋白多肽类药物口服给药的研究进展[J].解放军药学学报, 2005,21(1).51-54.
[120]Ledger R,Tucker I G,Walker G F. The metabolic barrier of the lower intestinal tract of salmon to the oral delivery of protein and peptide drugs[J]. J Control elease, 2002, 85 (1-3):91-103.
[121]文爱东.吸收促进剂在口服制剂中增强药物生物利用度的作用[J].国外医学·药学分册,2000,27(6):354-357.
[122]黄敏,吴伟.蛋白多肽类口服给药系统[J].中国临床药学杂志,2004,13(3):183-185.
[123]王萍,陈钧.酶抑制剂在蛋白质和肽类药物口服制剂中的应用[J].中国医药工业杂志,2005,36(8):5l0-514.
[124]Carreno-Gómez B, Woodley J F, Florence A T. Studies on the uptake of tomato lectin nanoparticles in everted gut sacs [J]. Int J Pharm,1999,183(1):7-11.
[125]Pan Y, Li Y J, Zhao H Y,et a1.Bioadhesive polysaccharide in protein delivery system: chitosan nanoparticles improve the intestinal absorption ofinsulin in vivo [J]. Int JPharm, 2002, 249(1-2):139.
[126]梅兴国.生物技术药物制剂基础与应用[M].北京:化学工业出版社,2004:308.
[127] Harris D, Robinson J R. Drug delivery via the mucous membranes of the oral cavity[J]. J Pharm Sci,1992, 81(1):1-9.
[128]金秀英.基因工程干扰素a-1治疗单疱角膜炎的临床研究[J].中华眼科杂志,1992, 28(3):134.
[129]朱雪研,韦建华,韦汉燕,等.动物药中蛋白和多肽的研究与开发[J].广西中医学院学报,2005,8(1):68-71.
[130]刘协,李小宁,沈逵,等.蚯蚓提取物对血栓形成和血小板聚集作用的实验研究[J].江苏预防医学,1997(2):8-9.
[131]张莅峡,郭育芝,刘泓,等.中药水蛭中的微量元素和氨基酸的分析[J].沈阳药科大学学报,1991,8(3):172-174,199.
[132]王四旺,王晓娟,谢艳华,等.蚕宝素中化学成分分析及其抗肿瘤活性初探[J].西北药学杂志,1997(5):211-213.
[133]焦波,娄红祥,王东兴,等.蜈蚣提取物对小鼠精原细胞的作用[J].山东医科大学学报,1999,37(4):358.
[134]徐立,方泰惠,许惠琴,等.血红林蚂蚁粉抗炎、镇痛作用的实验研究[J].南京中医药大学学报,1997,13(4):22-25.
[135]高梅,窦肇华,王典瑞.大黑蚂蚁水提液对阳虚动物模型免疫功能的影响[J].中国老年学杂志,1999,19(1):49-50.
[136]梁兆祥,王典瑞,许国战.蚂蚁水提取物对小鼠免疫功能的影响[J].吉林军医学院学报,2000,22(3):130-132.
[137]潘汉杰,周少雄.海蛇乙醇浸出物对小鼠免疫系统的影响[J].蛇志,2001,13(2):1-3.
[138]陈晓东,林建华.鹿茸多肽对大鼠软骨细胞复制性老化的作用[J].中国骨伤,2008,21(7):515-518.
[139]李振华,赵文海,冷向阳,等.鹿茸多肽对兔骨性关节炎软骨细胞增殖及凋亡调节作用的实验研究[J].世界中西医结合杂志,2009(10):701-703.
[140]王志兵,邱芳萍,解耸林[J].鹿角盘蛋白多肽的制备与活性研究[J].中国食品学报,2008,8(3):28-32.
[141]柯李晶,林冬云,黄晓南,等.不同加工工艺鹿茸的蛋白成分和活性比较[J].中药材,2008(1):11-14.
[142]潘凤光,孙威,周玉,等.梅花鹿鹿茸活性多肽的提取及免疫功效的初步研究[J].中国生物制品学杂志,2007,20(9):669-673.
[143]孙晓迪.鹿筋胶原治疗类风湿性关节炎的药效学研究[D].长春:长春中医药大学,2009.
[144]高艳华,王联结.国内外食品蛋白质发展概况[J].食品研究与开发,2003,24(5):58-60.
[145]董文滨,杨兆艳,胡鲜丽,等.动物蛋白生物活性肽的研究进展[J].食品研究与开发,2004,25(5):66-69.
[145] Meisel H. Chemical characterization and opioid activity of an exorphin isolated from in vivo digests of casein [J]. FEBS Lett, 1986,196(2):223-227.
[146]Brantl V, Teschemacher H, Henschen A. et al. Novel opioid peptides derived from casein (beta-casomorphins). I. Isolation from bovine casein peptone [J]. Hoppe Seylers Z Physiol Chem, 1979, 360(9):1211-1216.
[147]张佳程.乳蛋白中的生物活性序列[J].食品与发酵工业,1996(6):54-57.
[148]Clare D A, Swaisgood H E. Bioactive milk peptides: A prospectus [Review]. J Dairy Sci, 200, 83(6):1187-1195.
[149]Chiba H, Tani F, Yoshikawa M. Opioid antagonist peptides derived from kappa-casein [J]. J Dairy Res, 1989,56(3):363-366.
[150]Richard J, Fitz G., Meisel H. Milk protein-derived peptide inhibitors of angiotensin-I-converting enzyme [J]. Br J Nutr, 2000,84(1):S33-S37.
[151]Kohmura M, Nio K,Kubo K, et al. Inhibition of Angiotensin-converting Enzyme by Synthetic Peptides of Humanβ-Casein (Biological Chemistry) [J]. Agric Biol Chem, 1989,53(8):2107-2114.
[152]Fiat A M, Daniele M S, Pierre J, et al. Biologically Active Peptides from Milk Proteins with Emphasis on Two Examples Concerning Antithrombotic and Immunomodulating Activities [J].J Dairy Sci,1993, 76(1):301-310.
[153]Lahov E, Regelson W. Antibacterial and immunostimulating casein-derived substances from milk: casecidin, isracidin peptides[J]. Food Chem Toxicol,1996,34:131-145.
[154]Jollès P, Lévy-Toledano S, Fiat A M, et al. Analogy between fibrinogen and casein. Effect of an undecapeptide isolated from kappa-casein on platelet function [J].Eur J Biochem, 1986, 158:379-382.
[155]Azuma N, Nagaune S, Ishino Y, et al. DNA-Synthesis Stimulating Peptides from Humanβ-Casein (Biological Chemistry) [J]. Agric Biol Chem, 1989, 53:2631-2636.
[156]Melo E P, Aires-Barros M R, Cabral J M. Reverse micelles and protein biotechnology[J]. Biotechnol Annu Rev, 2001(7):87-129.
[157]赵丽丽,王昌涛,何聪芬,等.动物蛋白在化妆品中的应用现状和前景展望[J].北京日化,2008(2):3-9.
[158]吴鹏,李平亚,李启洋.海星胶原蛋白成分及生物活性的研究进展[J].时珍国医国药,2006,17(7):1297.
[159]萝莉,叶元土,林仕梅,等.动物蛋白质降解研究进展[J].动物医学进展,2001,22(4):12-16.
[160]宋焕禄,廖国洪.动物蛋白酶解研究(Ⅰ)[J].食品科学,2001,22(5):21-26.
[161]宋焕禄,廖国洪.动物蛋白酶解研究(Ⅱ)[J].食品科学,2001,22(6):28-32.
[162]汤高奇,柳艳霞,杨继红.酶法水解动物蛋白工艺研究[J].杨凌职业技术学院学报,2004,3(4):6-7,17.
[163]钱俊青,何国庆.植物蛋白的开发利用[J].中国食物与营养,2000(2):23-26.
[164]蔡华珍.我国植物蛋白质资源加工利用状况[J].安徽农业技术师范学院学报,1999,13(3):67-72.
[165]李世成,王启荣,杨则宜,等.活性肽在大鼠小肠吸收特点的实验研究[J].中国运动医学杂志,2004,23(3):271-275.
[166]王启荣,李肃反,杨则宜,等.补充大豆多肽对中长跑运动员训练期生化指标的影响[J].中国运动医学杂志,2004,23(1):33-37.
[167]王世英.免疫功能活性肽[J].生物学通报,2001,36(1):5-7.
[168]Tsuruki T, Yoshikawa M. Design of soymetide-4 derivatives to potentiate the anti-alopecia effect [J]. Biosci Biotech Biochem, 2004,68(5):1139-1141.
[169]Yasuyuki T, Yoshikawa M. Introduction of enterostatin(VPDPR) and a related sequence into soybean proglycinin AlaBlb subunit by site directed mutagenesis [J]. Biosci BiotechBiochem, 2000, 64(12):2731-2733.
[170]赵秀娟,王小雪,吴博学,等.大豆活性肽粉对喂饲高脂饲料大鼠血脂的影响[J].中国卫生检验杂志,2002,12(4):421-422.
[171]Yang H Y, Yang S C, Chen J R, et a1. Soyabean protein hydrolysate prevents the development of hypertension in spontaneously hypertensive rats[J]. Brit J Nutr, 2004,92(3):507-512.
[172]Wu J P, Ding X. Hypotensive and physiological effect of angi otensin converting enzyme inhibitory peptides derived from soy protein on spon taneously hypertensive rats[J].J Agric Food Chem,2001,49(1):501-506.
[173]荣建华,李小定,谢笔钧.大豆肽抗氧化效果的研究[J].食品科学,2002,23(11):118-120.
[174]何慧,谢笔钧,扬卓,等.大豆蛋白和玉米蛋白酶解物及其活性研究[J].粮油食品科技,2002,10(1):14-16.
[175]黄莉,江连洲,朱秀清.大豆蛋白抗氧性肽的研究[J].大豆通报,2003(5):20-21.
[176]张莉莉,严群芳,王恬.大豆生物活性肽的分离及其抗氧化活性研究[J].食品科学,2007,28(5):208-210.
[177]左伟勇,孟婷,陈伟华.植物蛋白源活性肽研究进展[J].饲料博览,2007(1):22-24,25.
[178]吴建平.一种开发米糠蛋白的新方法[J].粮食与饲料工业,1997(10):33-34.
[179]王梅.酶解玉米黄粉蛋白制备可溶性肽[J].粮油食品科技,1999,7(1):1-3.
[180]杨小军,左伟勇,陈伟华,等.面筋蛋白的胃蛋白酶酶解物对大鼠免疫功能的影响[J].南京农业大学学报,2004,27(4):69-72.
[181]刘世会.杜仲蛋白纯化、序列分析及抗菌活性研究[D].贵阳:贵州大学,2008.
[182]刘振宇.海藻中两种与植物抗病相关铜结合蛋白的分离、特性和基因特征[D].福州:福建农林大学,2004.
[183]张艺.苜蓿叶蛋白抗氧化功能的实验研究[D].重庆:重庆医科大学,2005.
[184]高富红,孙健,吴国娟.天花粉蛋白抗肿瘤作用机理研究概括[J].动物医学进展,2005,26(11):5-7.
[185]Gstirnor F, Vogt H J. Poptidos in White Koroan Ginseng [J].Arch Pharm, 1966,299(11):934-937.
[186]Ando T, Muraoka T, Yamasaki N, et al. Preparation of anti-lipolytic substance fromPanax Ginseng[J]. Planta Med, 1980,38:18-23.
[187]张今,赵宗健.人参中氨基酸及多肽的研究[J].高等学校化学学报, 1985, 61(4):376-380.
[188]张今,杜文媛,张红缨,等.胰岛素样人参多肽的氨基酸序列测定[J].吉林大学自然科学学报,1988(4):75-78.
[189]汪大伟,张红缨,张今.人参肽的二级结构分析[J].生物化学与生物物理学报,1989,21(2):175-176.
[190]张今,张红缨,陈冬松,等.人参肽基因的化学合成与克隆[J].生物化学杂志,1990,6(3):193-195.
[191]张今,张红缨,滕利荣,等.人参种子DNA诱导小麦变异的研究—从人参种子DNA处理过的小麦幼苗鉴别出异样的蛋白质和多肽[J].吉林大学学报(理学版), 1989(1):109-112.
[192]Kim SⅡ, Lee C Y, Jo D H. Extraction and Purification of Ginseng Oligopeptides with Antilipolytic Activities [J]. Korea J thickening, 1987,30(1):88-94.
[193]Takaku T, Kameda K, Matsura Y. Studies on isulin-like substance in Korean red ginseng[J].Planta Med, 1990,56(1):27-30.
[194]王本祥,杨名,金玉莲,等.人参多肽的降血糖作用[J].药学学报,1990,25(6):401-402.
[195]王本祥,杨名,金玉莲,等.人参多肽降血糖机制的研究[J].药学学报, 1990, 25 (10):727-728.
[196]魏俊杰,李亚平.生晒参多肽的提取分离[J].白求恩医科大学学报, 1990, 16 (5):436-438.
[197]吴庆夫,魏俊杰,徐景达.红参中肽类成分的分离鉴定[J].药学学报, 1991,26(7):499.
[198]曹立亚,王友茹.冻干参与传统商品参化学成分的比较研究:Ⅳ人参蛋白质成分分析中草药,1991,22(11):496.
[199]侯元.孙萍,焦连庆,等.人参蛋白的等电聚焦电泳分析[J].人参研究,1992(4):34-36.
[200]朱苗力,佘平,张华.人参多肽最低构象能计算分析[J].吉首大学学报(自然科学),1992,13(6):20-24.
[201]陈冬松,阎冰.胰岛素样人参多肽基因的克隆[J].辽宁大学学报,1996,23(1):85-88.
[202]Kajiwara H, Hemmings A M , Hirano H. Evidence of metal binding activities ofpentadecapeptide from Panax ginseng [J]. J Chromatogr B, 1996,687(2):443-448.
[203]Chen Z K, Fan C X ,Ye Y H, et al. Isolation and characterization of a group of oligopeptides related to oxidized glutathione from the root Panax ginseng [J]. J Peptide Res, 1998,52 (2):137-142.
[204]Nam M H, Kim S I, Liu J R, et al. Proteomic analysis of Korean ginseng (Panax ginseng C.A. Meyer) [J]. J Chromatogr B, 2005, 815(1-2):147-155.
[205]Kim S, Kweon S M, Kim E A. Characterization of Rnase-like major storage protein from the ginseng root by proteomic approach[J]. J Plant Physiol, 2004,161(7):837-845.
[206]Zhang C Z, Yu H S, BaoY M, et al. Purification and characterization of Ginsenoside-β-Glucosidase from Ginseng[J]. Chem Pharm Bull, 2001,49(7):795-798.
[207]Ng T B, Wang H X. Panaxagin, a new protein from Chinese ginseng possesses anti-fungal, anti-viral, translation-inhibiting and ribonuclease activities[J]. Life Sci, 2001,68(7):739-749.
[208]Yoon J Y, Ha B H, Woo J S, et al. Purification and characterization of a 28-kDa major protein from ginseng root[J]. Comp Biochem Phys B, 2002,132(3):551-557.
[209]Wang H X, Ng T B. A ribonuclease from Chinese ginseng (Panax ginseng) flowers [J]. Protein Expres Purif, 2004,33(2):195-199.
[210]闫亚军.重组人参多肽的研究[D].北京:北京化工大学, 2002.
[211]腾宇,邱芳萍,严铭铭,等.人参多肽的分离纯化[J].长春工业大学学报, 2005, 26(3):181-183.
[212]马波,邱芳萍,严铭铭,等.人参蛋白的分离纯化[J].现代食品科技,2006,22 (4):156-157,163.
[213]Haasan H M, Fridovich I. Regulation of superoxide dismutase synthesis in Escherichia coli: glucose effect[J]. J Bacteriol, 1977,132(2):505-510.
[214]李向高,郑友兰,贾继红.吉林红参与高丽红参的比较研究(Ⅵ)—种人参中酶活性的测定[J].中药材,1986(4):40-41.
[215]杨白纯.人参皂甙抗心肌缺血与抗自由基的关系[J].中华医学杂志,1988,68 (5):294-295.
[216]储国祥,陈修.人参总皂甙对大鼠急性脑缺血/再灌注损伤的保护作用(英文)[J].中国药理学与毒理学杂志,1989,3(1):23.
[217]张敏.人参总皂甙对大鼠烟雾吸入性肺损伤的疗效与其抗氧化作用的关系[J].解放军药学学报,1989,11(3):183.
[218]徐琳,赵雪俭,赵丹,等.人参二醇皂甙对失血性休克犬单胺类递质和单胺氧化酶的影响[J].中国病理生理杂志,1990,6(2):69-72.
[219]胡刚,宗瑞义,陈声武.人参茎叶皂甙对离题兔脑钠、钾-三磷酸腺苷酶活力的影响[J].白求恩医科大学学报,1987,13(5):403-407.
[220]宗瑞义,胡刚,陈声武.人参二醇皂甙和三醇皂甙对兔纹状体ATP酶的影响[J].药学学报,1988,23(7):494-497.
[221]胡刚,宋瑞义.人参皂甙对犬心肌Na+·K+-ATP酶活力的影响[J].中国药理学通报,1990(1):46-48.
[222]郑玉群,曹瑾,刘天培,等.几种人参皂甙对大鼠肾脏微粒体Na+·K+-ATP酶的抑制作用[J].中国药理学与毒理学杂志,1990,4(2):142-145.
[223]睢大筼,吕忠智,于晓峰.复合人参素对老龄大鼠血清过氧化脂质和超氧化物歧化酶的影响[J].人参研究,1992(4):14-15.
[224]宋淑兰,谢启文,王玉波,等.人参茎叶皂甙对老龄大鼠脑超氧化物歧化酶活力的影响[J].中国应用生理学杂志,1992,8(1):94-95.
[225]吴春福.依据自由基学说研究人参茎叶皂甙的抗衰老作用[J].沈阳药学院学报,1992,9(1):37-40.
[226]杨世海,方阵,杨继祥.GA对人参种子形态后熟过程中几种酶活力的影响[J].人参研究,1993(3):14-15.
[227]果崇真,曹悦群,孙非.不同光质对人参过氧化物酶活性的影响[J].辽宁农业科学,1993(3):53-55.
[228]张予阳,于庆海,吴春福.人参茎叶皂甙及其单体对大鼠脑内B型单胺氧化酶活性的影响[J].沈阳药学院学报,1993,10(1):51-54.
[229]刘仁海,周翔,刘吉民,等.人参根、叶皂甙对大鼠血清抗氧化酶活性和过氧化脂质的影响[J].1993,18(3):176-177.
[230]王建,金文,沈映君.单味人参和定志小丸对小鼠血液中超氧化物歧化酶活力的影响[J].成都中医学院学报,1994,17(3):36-39.
[231]吕薇,杨义军,赵懿峰,等.人参多糖对冷适应大鼠红细胞膜Na+·K+-ATP酶活性的影响[J].沈阳部队医药,1995,8(1):44-45.
[232]郭颂,陈满秋,赵节绪,等.人参茎叶皂甙对阿霉素中毒小鼠过氧化脂质和超氧化物歧化酶的影响[J].白求恩医科大学学报,1996,22(3):239-240.
[233]张志娥,石思信,肖建平.人参种子后熟期间过氧化物酶同功酶的变化[J].特产研究,1996(1):3-5.
[234]董秀文,杨晓晖.延缓衰老中药实验研究近况[J].山东中医药大学学报,1997,21(4):315-318.
[235]曹军,杜日新,凌云,等.人参蜂王浆口服液对抗氧化酶活力的影响[J].齐齐哈尔医学院学报,1999,20(1):4-5.
[236]Lim J Y, Ishiguro K, Kubo I. Tyrosinase inhibitory p-Coumaric acid from Ginseng leaves[J]. Phytother Res,1999,13(5):371-375.
[237]张嘉麟,徐文安,杨荫康,等.三七中人参皂甙对老年鼠血液中抗氧化酶活力影响的研究[J].昆明医学院学报, 2000,21(2):63-65.
[238]卢葵花.人参皂甙对中分子物质抑制红细胞ATP酶活性作用的影响[J].咸宁医学院学报,2001,15(4):254-255.
[239]李晓林,陆艳娟,李晓梅,等.人参二醇皂甙对内毒素诱导休克大鼠模型肺组织超氧化物歧化酶和丙二醛水平的影响[J].中国临床康复,2006,10(15):123-125.
[240]张亚玉,宋晓霞,孙海,等.栽培人参、西洋参不同年限土壤酸碱度和酶活性的变化[J].特产研究,2008(4):34-36.
[241]张岩,金凤燮,鱼红闪.GS0202菌产人参皂苷糖苷酶的酶性质研究[J].食品与发酵工业,2009,35(4):16-19.
[242]吴梧桐.生物化学(第5版)[M].北京:人民卫生出版社,2004:139.
[243]Markert C L, Moller F. Multiple forms of enzymes: tissue, ontogenetic, and species specific patterns[J]. Proc Natl Acad Sci U S ,1959, 45(5):753–763.
[244]胡能书,万国贤.同工酶技术及应用[M].长沙:湖南科技出版社,1985:2-17.
[245]高昆.同工酶技术在植物生态学研究中的应用[J].山西大同大学学报(自然科学版) ,2009,25(2):62-65.
[246]李志忠,吴婷婷.同工酶技术及其应用研究[J].甘肃高师学报(自然科学版),1999,4(2):46-52.
[247]陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2002:239-245.
[248]萧浪涛,王三根.植物生理学实验技术[M].北京:中国农业出版社,2005:82-83.
[249]陈建勋,王晓峰.植物生理学实验指导(第2版)[M].广州:华南理工大学出版社,2002:72-73.
[250]张志良,吴光耀.植物生物化学技术和方法[M].北京:农业出版社,1986:40-42.
[251]张志良,瞿伟菁.植物生理学实验指导(第3版)[M].北京:高等教育出版社,2003: 121-122
[252]彭志英,蒋黎.紫外速率直接法测定过氧化氢酶活性[J].华西医学,1995,10(1):4-7.
[253]杨兰芳,庞静,彭小兰.紫外分光光度法测定植物过氧化物酶活力[J].现代农业科技,2009(20):364-362.
[254] GB/T5009?171-2003.保健食品中超氧化物歧化酶(SOD)活性的测定[S].
[255]俞建英,蒋宇,王善利.生物化学实验技术[M].化学工业出版社,2005:266-269.
[256]张志良,瞿伟菁.植物生理学实验指导(第3版)[M].北京:高等教育出版社, 2002:135-136.
[257]翁霞,辛广,孟玲.小麦酯酶的纯化研究[J].鞍山师范学院学报,2008,10(4):31-34.
[258]王談,钱士匀.生物化学和临床生物化学检验[M].北京:清华大学出版社,2005: 152-155.
[259]余叔文,汤章城.植物生理学与分子生物学(第2版)[M].北京:科学出版社,1998:366 - 389.
[260]蒋明义.水分胁迫下植物体内的OH产生与细胞的氧化损害[J].植物学报,1999,41 (3):229 - 234.
[261]辛树权,赵骥民,何正飚,等.促生菌对黄瓜幼苗过氧化物酶活力的影响[J].长春师范学院学报(自然科学版),2008,27(1):59-62.
[262]张健,刘美艳.土人参的抗氧化成分分析[J].江苏农业科学,2005(1):109-110.
[263]杨淑慎,高俊凤.活性氧、自由基与植物的衰老[J].西北植物学报,2001,21 (2):215– 220
[264]张志娥,石思信,肖建平,等.人参种子后熟期间过氧化物酶同工酶的变化[J].特产研究,1996(1):3-5.
[265]Bonfill M, CusidóR M, Palazón J, et al. Relationship between peroxidase activity andorganogenesis in Panax ginseng calluses[J]. Plant Cell, Tissue and Organ Culture, 2003,73(1):37-41.
[266]胡卓逸,史建勋,丁家宜.人参细胞培养物中过氧化氢酶活力的测定[J].中草药, 1997,28(6):335-337.
[239]李晓林,陆艳娟,李晓梅,等.人参二醇皂甙对内毒素诱导休克大鼠模型肺组织超氧化物歧化酶和丙二醛水平的影响[J].中国临床康复,2006,10(15):123-125.
[267]汪少云,叶秀云,饶平凡.绿豆中苹果酸脱氢酶的性质及生物进化的研究[J].中国食品学报,2006,6(1):262-266.
[268]李洪山,张晓岚,周培之.旱生植物梭梭苹果酸脱氢酶的纯化及其性质的研究[J].新疆大学学报(自然科学版),1994,11(1):72-76.
[269]王以柔,李平,刘鸿先,等.低温对不同耐寒力的黄瓜幼苗子叶各细胞器中NAD+—苹果酸脱氢酶的影响[J].植物生理学报,1985,11(2):147-154.
[270]刘珠耀,范惠琴,王海庭.番茄苹果脱氢酶同工酶分析[J].植物生理通讯,1983,(2):23-26.
[271]刘琳,肖光辉,肖晓玲.酯酶和苹果酸脱氢酶同工酶分析用于香菇原种的鉴定[J].湖南农业科学,2000(3):51-53.
[272]马国龙,孙伟,常洪,等.洼地绵羊苹果酸脱氢酶同工酶研究[J].草食家畜,2006(3):5-7.
[273]丛辉,王惠民,王跃国,等.毛细管电泳法分离血清中乳酸脱氢酶同工酶[J].中华检验医学杂志,2005,28(11):1143-1145.
[274]宋鉴清,邹大伟,吕宁,等.乳酸脱氢酶(LDH)及其同工酶对良、恶性浆膜腔积液的鉴别诊断价值[J].山东医药,2009,49(3):68-69.
[275]Pecina, P, Houstkova, H, Hansikova, H, et al. Genetic Defects of Cytocrhome c Oxidase Assembly[J]. Physiol Res, 2004,53 (Suppl. 1): S213-S223.
[276]郭晓霞,郑哲民,于广志.酯酶同工酶多态性及其在昆虫分类学中的应用价值[J].昆虫知识,2000,37(6):371-374.
[277]王建,郑军,徐志国.单味人参、定志小丸等对小鼠组织胆碱酯酶活力的影响[J].陕西中医学院学报,1994,17(4):37-39.
[278]吴应文.大麦芽中酸性磷酸酯酶的分离[J].甘肃科学学报,1991,3(4):69-71.
[279]陈禅友,万茜.长豇豆种子的酸性磷酸(酯)酶活性分析及其应用[J].江汉大学学报,1999,16(6):18-19.
[280]杨立红,孙振兴,王晓洁,等.刺参酸性磷酸酯酶的分离纯化及部分性质研究[J].食品科学,2008,29(10):440-443.
[281]Yamasaki K H, Yokoyama H, Miyano K Y, et al. Purification and Characterization ofβ-Amylase from Ginseng[J]. Chem Pharm Bull,1989,37(4):973-978.
[282]赵可伟,司徒儒如,巫剑雄,等.血清中葡萄糖6-磷酸异构酶在类风湿关节炎中的诊断价值[J].热带医学杂志,2008,8(8):813-815.
[283]李慧颖,姜国平,吕英春,等.葡萄糖6-磷酸异构酶在类风湿关节炎中的诊断意义[J].吉林医学,2009,30(11):961-963.
[284]刘胜群.人参规范化生产操作技术(GAP)[D].长春:吉林农业大学, 2003,1-6.
[285]姜先刚,赵雨,唐任能,等.鲜人参药材水提取物的高效凝胶过滤色谱指纹图谱研究[J].药物分析,2008, 28(8):1222-1225.
[286]姜先刚,赵雨,张巍,等.人参水溶性蛋白SDS-聚丙烯酰胺凝胶电泳指纹图谱研究[J].药物分析,2008, 28(6):873-876.
[287]Soldati F, Tanaka O. pnnax ginseng: relation between age of Plant and content of gensenosides[J]. Planta Medica, 1984, 51(4):351-352.
[288]王世民.人参不同部位的皂甙组成及含量[J].国外医药﹒植物药分册,1981(5): 43.
[289]吴冬梅,屈海云,卢菲,等.端视电感耦合等离子体原子发射光谱法测定人参不同部位中的微量元素[J].分析科学学报,2007(5):563-566.
[290]王莉.人参与西洋参的比较鉴别[J].海峡药学,2007,19(4):58-59.
[291]陈燕.鲜人参、生晒参和红参的比较研究[J].海峡药学,2006,18(4):137-139.
[292]印敏芳.山参质量的经验鉴别[J].中国药业,2006,15 (9):56-57.
[293]楼余淦,凌明,程存归,等.生晒山参、西洋参、红参和生晒参的FTIR图谱研究[J].中草药,2003,34(7):665-667.
[294]柳良燕.人参、西洋参功效比较及用法研究[J].实用医技杂志,2008,15(34):41-43.
[295]姜海平,窦德强,荆淑琴,等.林下山参的人参皂苷含量分析和指纹图谱研究[J].中国现代中药,2008,10(4):12-15.
[296] Petros A J, Turner S C, Nunn A J. Cost implications of using inhaled nitric oxide compared with epoprostenol for pulmonary hypertension[J]. J Pharm Technol, 1995,11 (4):163-166.
[297]Hsieh M T, Peng W H, Wu C R, et al. The ameliorating effects of the cognitive- enhancing Chinese herbs on scopolamine-induced amnesia in rats[J]. Phytother Res, 2000, 14(5):375- 377.
[298]Wesnes K A, Ward T, McGinty A, et al. The memory enhancing effects of a Ginkgo biloba / Panax ginseng combination in healthy middleaged volunteers[J]. Psychopharmacology, 2000,152 (4):353-361.
[299]Gold J L, Laxer D A, Dergal J M, et al. Herbal drug therapy interactions: a focus on dementia[J]. Curr Opin Clin Nutr Metab Care, 2001, 4 (1): 29-34.
[300]Yoshikawa T, Asada Y, Furuya T. Continuous production of glycosides by a bioreactor using ginseng hairy root culture[J]. Appl Microbiolo Biotechnol, 1993, 39(4-5):460-464.
[301]Asada Y, Saito H, Yoshikawa T, et al. Biotransformation of 18 beta- glycyrrhet inic acid by ginseng hairy root culture[J]. phytochemistry, 1993,34(4):1049-1052.
[302]Gennady P M, Larisa I F, Yuri N Z, et al. Primary structures of two ribonucleases from ginseng calluses: New members of the PR-10 family of intracellular pathogenesis-related plant proteins. FEBS Lett, 1997,407(2):207-210.
[303]Abe1 F, Miura T, Nagahama1 T, et al. Isolation of a highly copper-tolerant yeast, Cryptococcus sp., from the Japan Trench and the induction of superoxide dismutase activity by Cu2+[J]. Biotechnology Lett, 2001,23(24):2027-2034.
[304]Bala K, Tripathy B C, Sharma D. Neuroprotective and anti-ageing effects of curcumin in aged rat brain regions[J]. Biogerontology, 2006,7(2):81-89.
[305]Kobayashi Y, Okahata S, Sakano T, et al. Superoxide dismutase activity of Tlym phocytes and non-T-lymphocytes[J]. FEBS Lett, 1979,98(2):391-393.
[306]Qu Y, Zhang H, Zhao S, et al. The effect on radioresistance of manganese superoxide dismutase in nasopharyngeal carcinoma[J]. Oncol Rep, 2010,23(4):1005-1011.
[307]Dees A, Zahl A, Puchta R, et al. Ivanovic′-Burmazovic′,I., Water exchange on seven-coordinate Mn(II) complexes with macrocyclic pentadentate ligands: insight in the mechanism of Mn(II) SOD mimetics[J]. Inorg Chem, 2007,46(7):2459-2470.
[308]许雅娟,赵艳景,胡虹.邻苯三酚自氧化法测定超氧化物歧化酶活性的研究[J].西南民族大学学报(自然科学版),2006,32(6):1207-1212.
[309]罗广华,王爱国.植物SOD的凝胶电泳及活性的显示[J].植物生理学通讯,1983 (6):44-45.
[310]郭尧军.蛋白质电泳实验技术(第1版)[M].北京:科学出版社,1993:54-56.
[311]Salin M L, Bridges S M. Isolation and Characterization of an Ironcontaining Superxide Dismutase from an Eucaryote Brassica Campestris[J]. Arch: J Biochem Biophys, 1980, 201(2):369-374.
[312]Kwiatowski J, Safianowska A, Kaniuga Z. Isolation and Characterization of an Ironcontaining Superxide Dismutase from Tomato Leaves[J]. Lycoersicon scalentum: Eur J Biochem, 1985,146(2):459-466.
[313]邹国林,罗时文,裘名宜.小白菜线粒体锰超氧化物歧化酶的纯化和性质研究[J].生物化学与生物物理学报,1992,24(2):183-183..
[314]McCord J M, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein)[J]. J Biol Chem, 1969,244(22):6049-6055.
[315]方允中,李文杰.自由基与酶基础理论及其在生物学医学中的应用[M].北京:科学出版社,1989:147-162.
[316]张彩莹.不同来源的超氧化物歧化酶部分理化性质比较研究[J].安徽农业科学,2007,35(15):4426-4429.
[317]王岚.超氧化物歧化酶的研究及应用概况[J].武汉工业学院学报.2002(1):36-40.
[318]Maral J, Puget K, Michelson A M. Comparative study of superoxide dismutase, catalase and glutathione peroxidase levels in erythrocytes of different animals[J]. Biochem Biophys Res Commun, 1977,77(4):1525-1535.
[319]Kobayashi Y, Okahata S, Sakano T, et al. Superoxide dismutase activity of T lymphocytes and non-T lymphocytes[J]. FEBS Lett, 1979,98(2):391-393.
[320]Cone R, Hasan S K, Lown J W, et al. The mechanism of the degradation of DNA by streptonigrin [J]. Can J Biochem, 1976,54(3):219-223.
[321]王琦,徐新莲,贾维峰,等.我国SOD水果生产现状与发展对策(续)[J].中国果业信息,2005(8):9-11.
[322]Manning M C, Patel K, Borchardt R T. Stability of protein pharmaceuticals[J]. Pharm Res, 1989,11(6):903-918.
[323]Arakawa T, Prest relsik S J, Kenney W C, et al. Factor affecting short - term and long– term stabilities of proteins[J]. Adv Drug Deli Rev, 2001,46(1-3):307-326.
[324]Fagain C O. Understanding and increasing protein stability[J]. Bioch Biophy Acta,1995, 1252(6):1-14.
[325] Chi E Y, Krishnan S, Randolph T W, et al. Physical stability of proteins in aqueous solution:mechanism and driving forces in nonnative protein aggregation[J]. Pharm Res, 2003,20(9):1325-1336.
[326]曲春香,沈颂东,王雪蜂,等.用考马斯亮蓝测定植物粗提液中可溶性蛋白质含量方法的研究[J].苏州大学学报(自然科学版),2006,22(2):82.
[327]施东魁,胡春梅.膜分离技术及其在医药生产和研究中的应用[J].中国中药杂志,2006,31(15):1221-1224.
[328]Wang H X, Ng T B. A ribonuclease from Chinese ginseng (Panax ginseng) flowers[J]. Protein Expres Purif, 2004,33(2):195-199.
[329]李红艳.人参水溶性蛋白及鹿茸胶原蛋白的活性研究[D].长春:长春中医药大学,2007.
[330]边兴艳,尹学念.MTT比色法及其应用[J].国外医学?临床生物化学与检验学分册, 1998,19(2):83-85.
[331]陈哲生.噻唑蓝比色法检测细胞的活性[J].上海医学检验杂志,1994,9(4):205-206.
[332]陈哲生.噻唑蓝比色法用来研究人类淋巴细胞和小鼠巨噬细胞的活性[J].上海免疫学杂志,1992,12(5):54-56.
[333]刘郁,刘连新.人参功效再认识[J].时珍国医国药,2006,17(2):289.
[334]蒋景华.人参的药理作用和临床应用[J].现代中西医结合杂志,2004,13(7):956-957.
[335]许见春.人参在实际应用中的药理作用[J].时珍国医国药,2006,17(4):578.
[336]王卫霞,王谦.人参皂苷的免疫调节作用极其应用[J].中华中医药杂志,2005,20 (4):234-236.
[337]梅少林,袁红艳,常雅萍,等.人参皂苷Rg3体外抗HSV-1活性与免疫调节效应[J].吉林大学学报(医学版),2006,32(6):1019-1022.
[338]褚秀玲,苏建青,韦旭斌.人参皂苷免疫调节和抗病毒作用研究进展[J].中兽医医药杂志,2008(5):20-23.
[339]郭志廷.经分子修饰的人参皂苷的免疫调节作用及机理研究[D].吉林大学硕士学位论文,2006.
[340]张晓芬,任伯成.人参抗运动疲劳作用的实验研究[J].中医药学报,1995(6):29.
[341]冯毅翀,潘华山,赵自明,等.运动性疲劳大鼠中枢神经递质改变及人参皂甙Rb1和Re1抗疲劳的实验研究[J].湖北中医杂志,2009,31(3):5-7.
[342]赵文莉,张馨,王伟,等.人参茎叶总皂甙抗疲劳作用的体内实验研究[J].卫生研究, 2009,38(2):184-187.
[343]王斌,张声华,李晓莉,等.人参总皂甙的耐缺氧效应机理研究[J].食品科学,2002,23(8):270-272.
[344]姜正林,吴新明,金淑仪,等.人参与丹参的抗脑缺氧作用.中国应用生理学杂志,2000, 16 (3):201.
[345]杨世杰,吴涛,周鸣,等.人参茎叶总甙耐窒息性缺氧的实验研究[J].吉林大学学报(医学版),1992,18(2):118-120.
[346]冯立明,番华珍,李慧云,等.人参的抗氧化作用[J].中西医结合杂志,1987,7(5):288-290,262.
[347]张纯武,滕红林,柳献云,等.人参皂甙对强直性脊柱炎镇痛和抗炎作用的实验研究[J].浙江中医杂志,2005,40(9):402-403.
[348]刘振明,吕莉莉,张若英,等.人参对吗啡镇痛作用的影响[J].中药药理与临床,1996,23(6):23-24.
[349]邬蓉,高守红,辛海量.人参抗辐射损伤研究概况[J].药学实践杂志,2007,25(6):364-366,392.
[350]姜建华,张学进,王育丽.人参皂苷对60Co照射小鼠不良反应的影响[J].浙江中西医结合杂志,2005,15(11):682-683.
[351]Ivanova T, Han Y, Son H J, et a1. Antimutagenic effect of polysaccharide ginsan extracted from Panax ginseng [J]. Food Chem Toxieol, 2006, 44(4):517-521.
[352]金光辉,刘树铮,刘扬,等.人参组甙对紫外线辐射致皮肤角质形成细胞损伤的保护作用[J].中华放射医学与防护杂志,2002,22(6):420-421.
[353]刘丽波,孙晓玲,张海英,等.人参三醇组苷对小鼠骨髓细胞染色体辐射的防护作用[J].吉林大学学报,2002,28(2):l38-140.
[354]李校坤,林灼锋,黄亚东,等.人参三醇组甙抗辐射作用的研究[J].中药材,2002,25(11):805-808.
[355]刘丽波,李玲,刘鹏来,等.人参皂苷对雄性小鼠骨髓细胞染色体的辐射防护作用.白求恩医科大学学报,1997,4 (23) :348-350.
[356]GPT2-1.中药、天然药物急性毒性研究技术指导原则[Z].2005.
[357]中华人民共和国卫生部.保健食品检验与评价技术规范[Z].2003.
[358]苏丙凡,张智慧,商亚珍.灯盏花素抑制角叉菜胶诱导小鼠血栓形成[J].承德医学院学报,2009,26(1):8-9.
[359]辛晓明,王大伟,王远丽,等.杜仲多糖镇痛作用研究[J].现代中西医结合杂志, 2009, 18(5):487-488.
[360]舒畅,张延英,蔺兴遥,等.红花凝胶剂抗炎镇痛作用的实验研究[J].中国实验方剂学杂志,2009,15(1):72-73.
[361]袁伯俊,廖明阳,李波.药物毒理学实验方法与技术[M].北京:化学工业出版社(生物·医药出版分社),2007:200-201,204.
[362]李晓波,金鑫,李妍,等.植物类中药活性蛋白成分研究进展[J].中草药,2004,35(6) 706-708.
[363]付秀花.中草药组方对小鼠免疫功能的影响[D].南京农业大学硕士学位论文,2003.
[364]吴红星,杨剑婷.抗辐射剂的研究现状[J].放射免疫学杂志,2005,18(1):58-60.
[365]崔凤梅,聂继华,李建祥,等.黑风蜂胶抗辐射损伤的实验研究[J].工业卫生与职业病,2006,32(5):283-285.
[366]邓砚,覃丽佳,谭钧,等.茉莉花提取液对老龄小鼠的抗氧化作用[J].临床研究,2009,6(5):17-18.
[367]郦章安,吴春福.现代老年药学[M].北京:中国医药科技出版社,200:430-441.
[368]Giardino R, Giavaresi G, Fini M, et al. The role of different chemical modifications of superoxide dismu-tase in preventing a prolonged muscular ischemia/reperfusion injury[J]. Artif Cell Blood Sub, 2002,30(3):189-198.
[369]杨江涛,杨娟,杨江冰.刺梨多糖对衰老小鼠体内抗氧化能力的影响[J].营养学报, 2008, 30(4):407-409.
[370]朱伟,孙红光,朱迅.黄芩苷元对炎症反应的影响[J].中国药理学通报,2009,25(2):194- 197.
[2]周小红.人参只卖萝卜价,东北参为何难敌高丽参[J].农村百事通,2007(6):6-8.
[3] Liu M, Zhang J. Effects of ginsenoside Rb1 and Rg1 on synaptosomal free calcium level, ATPase and calmodulin in rat hippocampus [J]. Chin Med J (Engl), 1995,108(7): 544-547.
[4]吴柏林,徐祝伟.人参及其复方制剂的抗衰老研究进展[J].时珍国医国药, 1997, 8(6): 560.
[5]程俊霖,周黎明,朱玲,等.人参茎叶总皂苷对衰老小鼠的作用研究[J].四川生理科学杂志,2004,26(3):97-99.
[6]崔巍,赵洪艳,王燕嬉.人参皂甙抗衰老的研究进展[J].中国老年学杂志,2006,26(11):1578-1581.
[7]徐承水.人参抗衰老作用的实验研究[J].赣南师范学院学报,2000(3):54-56.
[8]陈冠敏,陈小萍,何聆,等.人参片对大鼠血脂水平及延缓衰老作用的实验研究[J].实验预防医学,2002,9(4):311-313.
[9]王红丽,吴铁,吴志华,等.人参皂甙抗皮肤衰老作用实验研究[J].广东药学院学报,2003,19(1):25-27.
[10]黄宗锈,陈冠敏,林蔚.人参皂甙延缓衰老的实验研究[J].实用预防医学,2001,8(5):385.
[11]潘鑫鑫,郑卫红,傅万玉,等.人参总皂甙抗衰老作用的实验研究[J].中国老年学杂志,1995,15(5):288-290.
[12]李凤人.人参挥发油对肿瘤的作用[C].国际人参研讨会论文集,长春,1992,179.
[13]高峰.人参与人参皂苷Rh2-恶性肿瘤治疗的新观点[J].人参研究,2001,13(4): 17-18.
[14]陶丽华,高峰,刘红岩,等.20(R)-人参皂苷Rh2抗癌细胞转移实验研究[J].人参研究,2002,14(4):17-18.
[15]张仲苗,江波,郑筱祥.人参皂苷Rg3对肿瘤放疗患者外周血淋巴细胞的体外免疫增强作用[J].中国药学杂志,2004,39(4):261-264.
[16] Huang D. Saponins and sapogenins for use in combination therapy for cancer [P]. U S,WO/2004/056379, 2004-07-08.
[17]刘忞.人参皂甙Rg1抗衰老和促智作用及其机制研究[J].生理科学进展, 1996, 27(2): 139-142.
[18]刘单,张均田.人参皂苷Rg_1对老年大鼠行为活动的改善作用[J].中国药学杂志, 1996,31(8):464-467.
[19]胡圣望,胡勇,胡王平.人参皂甙Rg1对慢性应激大鼠空间学习记忆能力的影响[J].四川中医,2004,22(3):14-15,16.
[20]沈仲理.人参对人体增强免疫力和抗病能力的研究[J].上海预防医学杂志, 1995, 7 (12):529-530.
[21]吴浩,林洪生,裴迎霞,等.人参皂甙Rg3对荷瘤及环磷酰胺化疗小鼠粘膜免疫力影响[J].中国肿瘤,2006,15(6):369-371.
[22]武淑芳,睢大员,于晓风,等.西洋参叶20s-原人参二醇组皂苷抗实验性心肌缺血作用及其机制[J].中国药学杂志,2002,37(2):100-103.
[23]田建明,李浩,叶金梅,等.人参皂苷Rg2对大鼠化学性心肌缺血的影响[J].中国中药杂志,2003,28(12):1191-1192.
[24]张秋梅,张喆,于德民,等.人参糖肽治疗2型糖尿病的临床观察[J].中国现代医学杂志, 2003,13(6):59,62.
[25] Kim D H, Bae E A, Han M J, et al. Novel use of the extract of processed ginseng and saponin isolated therefrom [P]. U S, 20050232908, 2005-10-20.
[26]金慧,周经纬,娄子恒.人参产业与区域经济发展[J].人参研究,2004,16(1):2-6.
[27]高涛,王立莹.我国人参市场现状及前景展望[J].人参研究,2006, (3):5-6.
[28]李向高.人参产业的创新战略[J].吉林农业大学学报,2002,24(5):1-5.
[29]张中朋,刘张林.人参出口额快速增长西洋参进口额高于出口—2008年1~6月份人参、西洋参进出口情况分析[J].中国现代中药,2008,10(8):35,44.
[30]戴昀弟.人参产业可持续发展对策探讨[J].社会科学战线·东北经济研究, 2007(5): 68- 70.
[31]孙峥,周滋养.吉林省人参产业发展探析[J].安徽农业科学,2008,36(23):10019-10021
[32]于志斌,罗扬.人参国际市场分析与出口形势预测[J].中国现代中药,2009,11(7):45,50.
[33]曹广成.中韩人参价值对比研究[J].中国科技博览,2008(24):251-253.
[34]张中朋,刘张林.2007年我国人参出口形势分析及展望[J].中国现代中药, 2008, 10(3): 44-45.
[35]屈文平.吉林省人参生产与出口情况的调研报告[J].对外经贸统计,2005(5):38-41.
[36]李刚,蔡天智.浅谈人参出口问题[J].中药研究与信息,2003,5(8):59-61.
[37]那中凯.国际市场上人参商战策略初探[J].中国医药情报,1999,5(3):166-170.
[38]杨朝丹,郭庆海.吉林省人参加工业发展探析[J].沈阳农业大学学报:社会科学版, 2007, 9(1):16-19.
[39]杨朝丹,丛媛媛.吉林省人参加工企业发展现状、问题及对策研究[J].商情, 2009 (24):33,71.
[40]赵文洋,孙琛.吉林省人参龙头企业发展策略探析[J].现代商业,2009(30):56-57.
[41]仲伟同,迟美丽.人参产业发展的根本出路在于药食同源[J].人参研究,2009(1):26-27.
[42]金玉姬.延边人参加工业存在的问题及对策[J].农村经济与科技,2008,19(12):65,68
[43] Barnes T, Goodacre J, Moots R J, et a1. Therapeutic targeting of cytokines [J].Cytokine, 2006,35(3/4):217-219.
[44]蒋建平.我国蛋白质的综合开发利用[J].中国食物与营养,1995(1):9-12
[45]马静,葛熙,昌增益.蛋白质功能研究:历史、现状和将来[J].生命科学,2007,19(3):294-300.
[46]查锡良.医学分子生物学[M].北京:人民卫生出版社,2003:245-299.
[47]催凌飞,王遂.蛋白质及其水解物的分析应用[J].哈尔滨商业大学学报,2002, 18 (1): 117-120.
[48] Cavazzana-Calvo M, Thrasher A, Mavilio F. The future of gene therapy: Balancing the risks and benefits of clinical trials [J]. Nat, 2004, 427:779-781.
[49] Mathe C, Sagot M F, Schiex T, eta1. Current methods of gene prediction, their strengths and weaknesses[J]. Nucleic Acids Res, 2002, 30(19):4103-4117.
[50] Blattner F R, Plunkett III G, Bloch C A, et a1. The complete genome sequence of Escherichia coli K-12 [J]. Science, 1997, 277(5331):1453-1474.
[51] Wood V, Gwilliam R, Rajandream M A, eta1. Thegenome sequence of Schizosaccharo myces pombe[J]. Nature, 2002, 415(6874):871-880.
[52] Waterstone R. Genome sequence of the nematode C. elegans: a platform for investingating biology[J]. Science, 1998, 282 (5396): 2012- 2018.
[53] Adams M D, Celniker S E, Holt R A, et a1. The genome sequence of Drosophila melanogaster[J]. Science, 2000, 287:2185-2195.
[54] Arabidopsis Genome Initiative. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana[J]. Nature, 2001,408(6814):796-815.
[55] Venter J C, Adams M D, Myers E W, et al. The sequence of the human genome[J]. Science, 2001, 291(5507):1304-1351.
[56]阚欢,李贤忠,陆斌.辣木叶蛋白质提取工艺研究[J].西部林业科学,2007,36(1):106-107.
[57]张晓辉,章克昌.非水溶性茶蛋白的提取工艺研究[J].食品研究与开发,2005,26 (3):64—67.
[58]刘香萍,李国良,王凤军.酸热法提取苜蓿叶蛋白初探[J].实验研究, 2007(1):10-11.
[59]刘高强,周虎,魏美才.稀盐法提取马尾松毛虫蛹中的蛋白质[J].食品科技, 2007, (9): 132-134.
[60]刘朋,邱芳萍.梅花鹿胎衣中蛋白质的提取分离[J].现代食品科技,2006,22(4):153-155.
[61]左玉萍,冯成利,党蕊叶.猕猴桃蛋白酶提取的工艺技术研究[J].西北农业学报, 2006, 15(6):127-129.
[62]张巍,李红艳,马晶,等.人参水溶性蛋白的纯化工艺研究[J].吉林农业大学学报, 2008, 30(1):36-39.
[63]李红艳,赵雨,张巍,等.人参水溶性蛋白对几种细胞增殖的影响[J].吉林农业大学学报,2008,30(5):705-707,711.
[64]唐任能,赵雨,孙晓迪,等.梅花鹿鹿茸、鹿托盘及鹿骨水溶性总蛋白比较研究[J].吉林中医药,2008,28(4):295-296.
[65]幺宝金.人参蛋白的提取分离及性质研究[D].长春:长春中医药大学,2009.
[66]孟林,田景振.动物组织蛋白质提纯方法的研究进展[J].食品与药品,2009,11(5):53-55.
[67] Damerval C, Vienne D D, Zivy M, et a1. Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins [J]. Electrophoresis,1986,7(1):52-54.
[68] Wu F S, Wang M Y. Extraction of proteins for sodium dodecyl sulfate-polyacrylamide gel electrophoresis from protease-rich plant tissues[J]. Anal Biochem. 1984(139):100-103.
[69] Nandaknmar M P, Shen J, Raman B, et a1. Solubilization of trichloroacetic acid (TCA) precipitated microbial proteins via naOH for two-dimensional electrophoresis[J]. J Proteome Res, 2003,2(1):89-93.
[70] Lee S J, Oh P S, Ko J H, et a1. Glycoprotein isolated from Gardenia jasminoides Ellis has a scavenging activity against oxygen radicals and inhibits the oxygen radical-linduced protein kinase C alpha and nuclear factor-kappa B in NIH/3T3 cells[J]. Environ Toxicolo Phar, 2006, 21(1):8-21.
[71]胡兴昌,唐仕华.板蓝根生药中蛋白质提取方法的比较[J].上海师范大学学报, 2004 (12):66-68.
[72]郭雪松,黄小杰,王烁,等.醇法大豆浓缩蛋白提取工艺的优化[J].食品工业科技, 2007, 28 (5):181-182.
[73] G?rg A, Weiss W, Dunn M J. Current two-dimensional electrophoresis technology for proteomics [J]. Proteomics, 2004, 4(12):3665-3685.
[74]王川,李燕,马志英,等.几种酶法从猪皮中提取胶原蛋白的对比研究[J].食品科学, 2007,28(1):201-204.
[75]沈莲清,黄光荣,王向阳,等.茶渣中蛋白质酶法提取工艺[J].食品与生物技术学报, 2006,25 (6):7-12.
[76]陈凤莲,方桂珍.不同方法分离小麦麸皮中蛋白质的研究[J].粮食加工, 2009, 34(4): 28-30,35.
[77]李二凤,何小维,罗志刚.胶原蛋白的提取工艺研究[J].食品研究与开发,2006, 27 (3): 64-65,63.
[78] Moulton K J, Wang L C. A pilot plant study ofcontinuous ultrasonic extraction of soybean protein [J]. J Food Sci, 1982,47(4):1127-1129.
[79]刘婷婷,王海灵,张艳荣.玉米超氧化物歧化酶超声波提取技术的研究[J].食品科学, 2007,28(8):156-159.
[80]龚丽芬,龚文贤.双水相分离提取胃蛋白酶和琼脂糖的初步研究[J].化学与生物工程,2006,23(11):36-38.
[81]赵晓燕,陈复生,薛文通.不同反胶束体系提取大豆蛋白质的研究[J].食品科学, 2007,28(4):109-111,112.
[82]李建武,萧能愿,余瑞元,等.生物化学实验原理和方法[M].北京:北京大学出版社, 1994: 160-176.
[83]郭颖娜,孙卫.蛋白质含量测定方法的比较[J].河北化工,2008,31(4):36-37.
[84]杨玉芳.蛋白质含量测定方法[J].明胶科学与技术,2007,27(2):99.
[85]薛志欣,杨桂朋,马晓梅,等.分光光度法测定藻胆蛋白含量的研究[J].鲁东大学学报(自然科学版),2008,24(3):250-253.
[86]康东周,张善玉,全仙花.Lowry法测定官颈癌片中蛋白质的含量[J].现代医药卫生, 2005,21(23):3210-3211.
[87]陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2002:166-167.
[88]曹艳,黄必胜.半夏总蛋白含量的紫外吸收法测定[J].湖北中医杂志,2005,27(7):48.
[89]王淡兮,孙秀兰.蛋白质定量检测方法的探讨[J].粮食与食品工业,2009,14(4):49-51.
[90] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantifies of protein utilizing the principle of protein-dye binding [J]. Anal Biochem, 1976,7(72):248-254.
[91]曲春香,沈颂东,王雪蜂,等.用考马斯亮蓝测定植物粗提液中可溶性蛋白质含量方法的研究[J].苏州大学学报(自然科学版),2006,22(2):82.
[92]汪良驹,姜卫兵,祝海阔,等.银杏叶片可溶性蛋白质测定非直线性原因分析[J].果树学报,2002,19(1):32.
[93]刘雅超,刘旭,白鸿源,等.蛋白质定量分析的研究进展[J].中国民族民间医药, 2009 (8): 49-50.
[94]王超英,周如真,戴国华,等.胸腺肽注射液分子量测定[J].中国生化药物杂志, 2000, 21(1):33-34.
[95] Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis[J]. J Biol Chem, 1969, 244 (16):4406-4412.
[96] Swank R T, Munkres K D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem, 1971, 39(2):462-477.
[97]崔韶晖,王艳颖,崔杰,等.大连湾牡蛎蛋白的分离提取及分子量测定[J].大连民族学院学报,2005,7(1):53-54,73.
[98]陈振江,殷丹,曹艳,等.厚朴种子蛋白质成分分子量测定[J].中成药, 2005, 27(6): 711- 712.
[99]陈绍农,潘远江,陈耀祖.多肽及蛋白质质谱分析新进展[J].质谱学报,1995, 16 (3): 15-21.
[100]Werner E, Bo U R, Sundqvist P, et al. Directional correlation between the primary particle and ejected molecular ions in electronic sputtering of large organic molecules [J]. Phys Rev B, 1989, 39(1):763-766.
[101]解建勋,蒲小平,李玉珍,等.蛋白质组分析技术进展[J].生物物理学报,2001,17:19 -26.
[102]Zhao S K, Somayajula K V, Sharkey A G, et al. Novel method for matrix-assisted laser mass spectrometry of proteins [J]. Anal Chem,1991,63(5):450-453.
[103]赵善楷,钟峰,张晓红.基体辅助激光解析质谱法(MALDI-MS)在生物化学中的应用[J].质谱学报,1996,17(2):1-7.
[104]刘慧敏,赖志辉,黎军,等.碎片结构分析在MALDI TOF MS法测定多肽序列中的应用[J].生物化学与生物物理学报, 2000 (32):179-182.
[105]郭元平,王书臻,曹振岭,等.榆叶梅果仁蛋白质的提取研究[J].农业与技术,2006, 26 (2) :88-89.
[106]0' Farrell P. High resolution two-dimensional electrophoresis of proteins[J]. J Biol Chem, 1975,250(10):4007-4021.
[107]Cordwell S J, Nouwens A S, Verrills N M, et al. Subproteomics based upon protein cellular location and relative solubilities in conjunction with composite two-dimensional electrophoresis gels[J]. Electrophoresis,2000, 21(6):1094-1103.
[108]Wang W, Scali M, Vignani R, et a1. Protein extraction for two-dimensional electrophoresis from olive leaf, a plant tissue containing high levels of interfering compounds [J]. Electrophoresis,2003,24(14):2369-2375.
[109]Zhao C F, Wang J Q, Cao M L, et a1. changes in rice leaves during development of field-grown rice plants[J]. Proteomics, 2005,5(4):961-972.
[110]安娜,赖钟雄,郭志雄.龙眼体细胞胚胎发生过程中特异表达蛋白的双向电泳分析[J].福建农林大学学报(自然科学版),2007,36(3):244-249.
[111]乐寅婷,李梅,陈倩,等.油菜叶片蛋白质组对机械损伤应答的初步分析[J].植物生态学报,2008,32(1):220-225.
[112]周蕴薇,董文珂,刘艳萍,等.山茶叶片可溶性蛋白双向电泳技术的建立[J].生物技术通报,2008(2):127-132.
[113]范海延,陈捷,张春宇,等.适于黄瓜叶片蛋白质组分析的双向电泳方法最佳条件的研究[J].沈阳农业大学学报,2008,39(3):365-367.
[114]罗泽宇,杨粤军,刘选明.拟南芥蛋白质组研究中双向电泳技术条件的优化[J].激光生物学报,2008,l7(4):539-543.
[115]古卓良,周国华,张晓丹,等.缓冲液种类对毛细管区带电泳分离蛋白质的影响[J].解放军药学学报,2003,19(6):404-407.
[116]朱京妃,夏明.毛细管电泳技术在多肽和蛋白质检测中的应用[J].农产品加工?创新版, 2009(6):54-56.
[117]Colao A, Somma C D, Salerno M, et a1. The Cardiovascular Risk of GH-Deficient Adolescents [J]. J Clin Endocrinol Metab, 2002, 87(8): 3650- 3655.
[118]孙晓东,房泽海,任悦欣.蛋白多肽类药物给药途径及剂型的研究进展①[ J].中外医疗,2009,21(7):160-162.
[119]金城,王海涛,肖小河.蛋白多肽类药物口服给药的研究进展[J].解放军药学学报, 2005,21(1).51-54.
[120]Ledger R,Tucker I G,Walker G F. The metabolic barrier of the lower intestinal tract of salmon to the oral delivery of protein and peptide drugs[J]. J Control elease, 2002, 85 (1-3):91-103.
[121]文爱东.吸收促进剂在口服制剂中增强药物生物利用度的作用[J].国外医学·药学分册,2000,27(6):354-357.
[122]黄敏,吴伟.蛋白多肽类口服给药系统[J].中国临床药学杂志,2004,13(3):183-185.
[123]王萍,陈钧.酶抑制剂在蛋白质和肽类药物口服制剂中的应用[J].中国医药工业杂志,2005,36(8):5l0-514.
[124]Carreno-Gómez B, Woodley J F, Florence A T. Studies on the uptake of tomato lectin nanoparticles in everted gut sacs [J]. Int J Pharm,1999,183(1):7-11.
[125]Pan Y, Li Y J, Zhao H Y,et a1.Bioadhesive polysaccharide in protein delivery system: chitosan nanoparticles improve the intestinal absorption ofinsulin in vivo [J]. Int JPharm, 2002, 249(1-2):139.
[126]梅兴国.生物技术药物制剂基础与应用[M].北京:化学工业出版社,2004:308.
[127] Harris D, Robinson J R. Drug delivery via the mucous membranes of the oral cavity[J]. J Pharm Sci,1992, 81(1):1-9.
[128]金秀英.基因工程干扰素a-1治疗单疱角膜炎的临床研究[J].中华眼科杂志,1992, 28(3):134.
[129]朱雪研,韦建华,韦汉燕,等.动物药中蛋白和多肽的研究与开发[J].广西中医学院学报,2005,8(1):68-71.
[130]刘协,李小宁,沈逵,等.蚯蚓提取物对血栓形成和血小板聚集作用的实验研究[J].江苏预防医学,1997(2):8-9.
[131]张莅峡,郭育芝,刘泓,等.中药水蛭中的微量元素和氨基酸的分析[J].沈阳药科大学学报,1991,8(3):172-174,199.
[132]王四旺,王晓娟,谢艳华,等.蚕宝素中化学成分分析及其抗肿瘤活性初探[J].西北药学杂志,1997(5):211-213.
[133]焦波,娄红祥,王东兴,等.蜈蚣提取物对小鼠精原细胞的作用[J].山东医科大学学报,1999,37(4):358.
[134]徐立,方泰惠,许惠琴,等.血红林蚂蚁粉抗炎、镇痛作用的实验研究[J].南京中医药大学学报,1997,13(4):22-25.
[135]高梅,窦肇华,王典瑞.大黑蚂蚁水提液对阳虚动物模型免疫功能的影响[J].中国老年学杂志,1999,19(1):49-50.
[136]梁兆祥,王典瑞,许国战.蚂蚁水提取物对小鼠免疫功能的影响[J].吉林军医学院学报,2000,22(3):130-132.
[137]潘汉杰,周少雄.海蛇乙醇浸出物对小鼠免疫系统的影响[J].蛇志,2001,13(2):1-3.
[138]陈晓东,林建华.鹿茸多肽对大鼠软骨细胞复制性老化的作用[J].中国骨伤,2008,21(7):515-518.
[139]李振华,赵文海,冷向阳,等.鹿茸多肽对兔骨性关节炎软骨细胞增殖及凋亡调节作用的实验研究[J].世界中西医结合杂志,2009(10):701-703.
[140]王志兵,邱芳萍,解耸林[J].鹿角盘蛋白多肽的制备与活性研究[J].中国食品学报,2008,8(3):28-32.
[141]柯李晶,林冬云,黄晓南,等.不同加工工艺鹿茸的蛋白成分和活性比较[J].中药材,2008(1):11-14.
[142]潘凤光,孙威,周玉,等.梅花鹿鹿茸活性多肽的提取及免疫功效的初步研究[J].中国生物制品学杂志,2007,20(9):669-673.
[143]孙晓迪.鹿筋胶原治疗类风湿性关节炎的药效学研究[D].长春:长春中医药大学,2009.
[144]高艳华,王联结.国内外食品蛋白质发展概况[J].食品研究与开发,2003,24(5):58-60.
[145]董文滨,杨兆艳,胡鲜丽,等.动物蛋白生物活性肽的研究进展[J].食品研究与开发,2004,25(5):66-69.
[145] Meisel H. Chemical characterization and opioid activity of an exorphin isolated from in vivo digests of casein [J]. FEBS Lett, 1986,196(2):223-227.
[146]Brantl V, Teschemacher H, Henschen A. et al. Novel opioid peptides derived from casein (beta-casomorphins). I. Isolation from bovine casein peptone [J]. Hoppe Seylers Z Physiol Chem, 1979, 360(9):1211-1216.
[147]张佳程.乳蛋白中的生物活性序列[J].食品与发酵工业,1996(6):54-57.
[148]Clare D A, Swaisgood H E. Bioactive milk peptides: A prospectus [Review]. J Dairy Sci, 200, 83(6):1187-1195.
[149]Chiba H, Tani F, Yoshikawa M. Opioid antagonist peptides derived from kappa-casein [J]. J Dairy Res, 1989,56(3):363-366.
[150]Richard J, Fitz G., Meisel H. Milk protein-derived peptide inhibitors of angiotensin-I-converting enzyme [J]. Br J Nutr, 2000,84(1):S33-S37.
[151]Kohmura M, Nio K,Kubo K, et al. Inhibition of Angiotensin-converting Enzyme by Synthetic Peptides of Humanβ-Casein (Biological Chemistry) [J]. Agric Biol Chem, 1989,53(8):2107-2114.
[152]Fiat A M, Daniele M S, Pierre J, et al. Biologically Active Peptides from Milk Proteins with Emphasis on Two Examples Concerning Antithrombotic and Immunomodulating Activities [J].J Dairy Sci,1993, 76(1):301-310.
[153]Lahov E, Regelson W. Antibacterial and immunostimulating casein-derived substances from milk: casecidin, isracidin peptides[J]. Food Chem Toxicol,1996,34:131-145.
[154]Jollès P, Lévy-Toledano S, Fiat A M, et al. Analogy between fibrinogen and casein. Effect of an undecapeptide isolated from kappa-casein on platelet function [J].Eur J Biochem, 1986, 158:379-382.
[155]Azuma N, Nagaune S, Ishino Y, et al. DNA-Synthesis Stimulating Peptides from Humanβ-Casein (Biological Chemistry) [J]. Agric Biol Chem, 1989, 53:2631-2636.
[156]Melo E P, Aires-Barros M R, Cabral J M. Reverse micelles and protein biotechnology[J]. Biotechnol Annu Rev, 2001(7):87-129.
[157]赵丽丽,王昌涛,何聪芬,等.动物蛋白在化妆品中的应用现状和前景展望[J].北京日化,2008(2):3-9.
[158]吴鹏,李平亚,李启洋.海星胶原蛋白成分及生物活性的研究进展[J].时珍国医国药,2006,17(7):1297.
[159]萝莉,叶元土,林仕梅,等.动物蛋白质降解研究进展[J].动物医学进展,2001,22(4):12-16.
[160]宋焕禄,廖国洪.动物蛋白酶解研究(Ⅰ)[J].食品科学,2001,22(5):21-26.
[161]宋焕禄,廖国洪.动物蛋白酶解研究(Ⅱ)[J].食品科学,2001,22(6):28-32.
[162]汤高奇,柳艳霞,杨继红.酶法水解动物蛋白工艺研究[J].杨凌职业技术学院学报,2004,3(4):6-7,17.
[163]钱俊青,何国庆.植物蛋白的开发利用[J].中国食物与营养,2000(2):23-26.
[164]蔡华珍.我国植物蛋白质资源加工利用状况[J].安徽农业技术师范学院学报,1999,13(3):67-72.
[165]李世成,王启荣,杨则宜,等.活性肽在大鼠小肠吸收特点的实验研究[J].中国运动医学杂志,2004,23(3):271-275.
[166]王启荣,李肃反,杨则宜,等.补充大豆多肽对中长跑运动员训练期生化指标的影响[J].中国运动医学杂志,2004,23(1):33-37.
[167]王世英.免疫功能活性肽[J].生物学通报,2001,36(1):5-7.
[168]Tsuruki T, Yoshikawa M. Design of soymetide-4 derivatives to potentiate the anti-alopecia effect [J]. Biosci Biotech Biochem, 2004,68(5):1139-1141.
[169]Yasuyuki T, Yoshikawa M. Introduction of enterostatin(VPDPR) and a related sequence into soybean proglycinin AlaBlb subunit by site directed mutagenesis [J]. Biosci BiotechBiochem, 2000, 64(12):2731-2733.
[170]赵秀娟,王小雪,吴博学,等.大豆活性肽粉对喂饲高脂饲料大鼠血脂的影响[J].中国卫生检验杂志,2002,12(4):421-422.
[171]Yang H Y, Yang S C, Chen J R, et a1. Soyabean protein hydrolysate prevents the development of hypertension in spontaneously hypertensive rats[J]. Brit J Nutr, 2004,92(3):507-512.
[172]Wu J P, Ding X. Hypotensive and physiological effect of angi otensin converting enzyme inhibitory peptides derived from soy protein on spon taneously hypertensive rats[J].J Agric Food Chem,2001,49(1):501-506.
[173]荣建华,李小定,谢笔钧.大豆肽抗氧化效果的研究[J].食品科学,2002,23(11):118-120.
[174]何慧,谢笔钧,扬卓,等.大豆蛋白和玉米蛋白酶解物及其活性研究[J].粮油食品科技,2002,10(1):14-16.
[175]黄莉,江连洲,朱秀清.大豆蛋白抗氧性肽的研究[J].大豆通报,2003(5):20-21.
[176]张莉莉,严群芳,王恬.大豆生物活性肽的分离及其抗氧化活性研究[J].食品科学,2007,28(5):208-210.
[177]左伟勇,孟婷,陈伟华.植物蛋白源活性肽研究进展[J].饲料博览,2007(1):22-24,25.
[178]吴建平.一种开发米糠蛋白的新方法[J].粮食与饲料工业,1997(10):33-34.
[179]王梅.酶解玉米黄粉蛋白制备可溶性肽[J].粮油食品科技,1999,7(1):1-3.
[180]杨小军,左伟勇,陈伟华,等.面筋蛋白的胃蛋白酶酶解物对大鼠免疫功能的影响[J].南京农业大学学报,2004,27(4):69-72.
[181]刘世会.杜仲蛋白纯化、序列分析及抗菌活性研究[D].贵阳:贵州大学,2008.
[182]刘振宇.海藻中两种与植物抗病相关铜结合蛋白的分离、特性和基因特征[D].福州:福建农林大学,2004.
[183]张艺.苜蓿叶蛋白抗氧化功能的实验研究[D].重庆:重庆医科大学,2005.
[184]高富红,孙健,吴国娟.天花粉蛋白抗肿瘤作用机理研究概括[J].动物医学进展,2005,26(11):5-7.
[185]Gstirnor F, Vogt H J. Poptidos in White Koroan Ginseng [J].Arch Pharm, 1966,299(11):934-937.
[186]Ando T, Muraoka T, Yamasaki N, et al. Preparation of anti-lipolytic substance fromPanax Ginseng[J]. Planta Med, 1980,38:18-23.
[187]张今,赵宗健.人参中氨基酸及多肽的研究[J].高等学校化学学报, 1985, 61(4):376-380.
[188]张今,杜文媛,张红缨,等.胰岛素样人参多肽的氨基酸序列测定[J].吉林大学自然科学学报,1988(4):75-78.
[189]汪大伟,张红缨,张今.人参肽的二级结构分析[J].生物化学与生物物理学报,1989,21(2):175-176.
[190]张今,张红缨,陈冬松,等.人参肽基因的化学合成与克隆[J].生物化学杂志,1990,6(3):193-195.
[191]张今,张红缨,滕利荣,等.人参种子DNA诱导小麦变异的研究—从人参种子DNA处理过的小麦幼苗鉴别出异样的蛋白质和多肽[J].吉林大学学报(理学版), 1989(1):109-112.
[192]Kim SⅡ, Lee C Y, Jo D H. Extraction and Purification of Ginseng Oligopeptides with Antilipolytic Activities [J]. Korea J thickening, 1987,30(1):88-94.
[193]Takaku T, Kameda K, Matsura Y. Studies on isulin-like substance in Korean red ginseng[J].Planta Med, 1990,56(1):27-30.
[194]王本祥,杨名,金玉莲,等.人参多肽的降血糖作用[J].药学学报,1990,25(6):401-402.
[195]王本祥,杨名,金玉莲,等.人参多肽降血糖机制的研究[J].药学学报, 1990, 25 (10):727-728.
[196]魏俊杰,李亚平.生晒参多肽的提取分离[J].白求恩医科大学学报, 1990, 16 (5):436-438.
[197]吴庆夫,魏俊杰,徐景达.红参中肽类成分的分离鉴定[J].药学学报, 1991,26(7):499.
[198]曹立亚,王友茹.冻干参与传统商品参化学成分的比较研究:Ⅳ人参蛋白质成分分析中草药,1991,22(11):496.
[199]侯元.孙萍,焦连庆,等.人参蛋白的等电聚焦电泳分析[J].人参研究,1992(4):34-36.
[200]朱苗力,佘平,张华.人参多肽最低构象能计算分析[J].吉首大学学报(自然科学),1992,13(6):20-24.
[201]陈冬松,阎冰.胰岛素样人参多肽基因的克隆[J].辽宁大学学报,1996,23(1):85-88.
[202]Kajiwara H, Hemmings A M , Hirano H. Evidence of metal binding activities ofpentadecapeptide from Panax ginseng [J]. J Chromatogr B, 1996,687(2):443-448.
[203]Chen Z K, Fan C X ,Ye Y H, et al. Isolation and characterization of a group of oligopeptides related to oxidized glutathione from the root Panax ginseng [J]. J Peptide Res, 1998,52 (2):137-142.
[204]Nam M H, Kim S I, Liu J R, et al. Proteomic analysis of Korean ginseng (Panax ginseng C.A. Meyer) [J]. J Chromatogr B, 2005, 815(1-2):147-155.
[205]Kim S, Kweon S M, Kim E A. Characterization of Rnase-like major storage protein from the ginseng root by proteomic approach[J]. J Plant Physiol, 2004,161(7):837-845.
[206]Zhang C Z, Yu H S, BaoY M, et al. Purification and characterization of Ginsenoside-β-Glucosidase from Ginseng[J]. Chem Pharm Bull, 2001,49(7):795-798.
[207]Ng T B, Wang H X. Panaxagin, a new protein from Chinese ginseng possesses anti-fungal, anti-viral, translation-inhibiting and ribonuclease activities[J]. Life Sci, 2001,68(7):739-749.
[208]Yoon J Y, Ha B H, Woo J S, et al. Purification and characterization of a 28-kDa major protein from ginseng root[J]. Comp Biochem Phys B, 2002,132(3):551-557.
[209]Wang H X, Ng T B. A ribonuclease from Chinese ginseng (Panax ginseng) flowers [J]. Protein Expres Purif, 2004,33(2):195-199.
[210]闫亚军.重组人参多肽的研究[D].北京:北京化工大学, 2002.
[211]腾宇,邱芳萍,严铭铭,等.人参多肽的分离纯化[J].长春工业大学学报, 2005, 26(3):181-183.
[212]马波,邱芳萍,严铭铭,等.人参蛋白的分离纯化[J].现代食品科技,2006,22 (4):156-157,163.
[213]Haasan H M, Fridovich I. Regulation of superoxide dismutase synthesis in Escherichia coli: glucose effect[J]. J Bacteriol, 1977,132(2):505-510.
[214]李向高,郑友兰,贾继红.吉林红参与高丽红参的比较研究(Ⅵ)—种人参中酶活性的测定[J].中药材,1986(4):40-41.
[215]杨白纯.人参皂甙抗心肌缺血与抗自由基的关系[J].中华医学杂志,1988,68 (5):294-295.
[216]储国祥,陈修.人参总皂甙对大鼠急性脑缺血/再灌注损伤的保护作用(英文)[J].中国药理学与毒理学杂志,1989,3(1):23.
[217]张敏.人参总皂甙对大鼠烟雾吸入性肺损伤的疗效与其抗氧化作用的关系[J].解放军药学学报,1989,11(3):183.
[218]徐琳,赵雪俭,赵丹,等.人参二醇皂甙对失血性休克犬单胺类递质和单胺氧化酶的影响[J].中国病理生理杂志,1990,6(2):69-72.
[219]胡刚,宗瑞义,陈声武.人参茎叶皂甙对离题兔脑钠、钾-三磷酸腺苷酶活力的影响[J].白求恩医科大学学报,1987,13(5):403-407.
[220]宗瑞义,胡刚,陈声武.人参二醇皂甙和三醇皂甙对兔纹状体ATP酶的影响[J].药学学报,1988,23(7):494-497.
[221]胡刚,宋瑞义.人参皂甙对犬心肌Na+·K+-ATP酶活力的影响[J].中国药理学通报,1990(1):46-48.
[222]郑玉群,曹瑾,刘天培,等.几种人参皂甙对大鼠肾脏微粒体Na+·K+-ATP酶的抑制作用[J].中国药理学与毒理学杂志,1990,4(2):142-145.
[223]睢大筼,吕忠智,于晓峰.复合人参素对老龄大鼠血清过氧化脂质和超氧化物歧化酶的影响[J].人参研究,1992(4):14-15.
[224]宋淑兰,谢启文,王玉波,等.人参茎叶皂甙对老龄大鼠脑超氧化物歧化酶活力的影响[J].中国应用生理学杂志,1992,8(1):94-95.
[225]吴春福.依据自由基学说研究人参茎叶皂甙的抗衰老作用[J].沈阳药学院学报,1992,9(1):37-40.
[226]杨世海,方阵,杨继祥.GA对人参种子形态后熟过程中几种酶活力的影响[J].人参研究,1993(3):14-15.
[227]果崇真,曹悦群,孙非.不同光质对人参过氧化物酶活性的影响[J].辽宁农业科学,1993(3):53-55.
[228]张予阳,于庆海,吴春福.人参茎叶皂甙及其单体对大鼠脑内B型单胺氧化酶活性的影响[J].沈阳药学院学报,1993,10(1):51-54.
[229]刘仁海,周翔,刘吉民,等.人参根、叶皂甙对大鼠血清抗氧化酶活性和过氧化脂质的影响[J].1993,18(3):176-177.
[230]王建,金文,沈映君.单味人参和定志小丸对小鼠血液中超氧化物歧化酶活力的影响[J].成都中医学院学报,1994,17(3):36-39.
[231]吕薇,杨义军,赵懿峰,等.人参多糖对冷适应大鼠红细胞膜Na+·K+-ATP酶活性的影响[J].沈阳部队医药,1995,8(1):44-45.
[232]郭颂,陈满秋,赵节绪,等.人参茎叶皂甙对阿霉素中毒小鼠过氧化脂质和超氧化物歧化酶的影响[J].白求恩医科大学学报,1996,22(3):239-240.
[233]张志娥,石思信,肖建平.人参种子后熟期间过氧化物酶同功酶的变化[J].特产研究,1996(1):3-5.
[234]董秀文,杨晓晖.延缓衰老中药实验研究近况[J].山东中医药大学学报,1997,21(4):315-318.
[235]曹军,杜日新,凌云,等.人参蜂王浆口服液对抗氧化酶活力的影响[J].齐齐哈尔医学院学报,1999,20(1):4-5.
[236]Lim J Y, Ishiguro K, Kubo I. Tyrosinase inhibitory p-Coumaric acid from Ginseng leaves[J]. Phytother Res,1999,13(5):371-375.
[237]张嘉麟,徐文安,杨荫康,等.三七中人参皂甙对老年鼠血液中抗氧化酶活力影响的研究[J].昆明医学院学报, 2000,21(2):63-65.
[238]卢葵花.人参皂甙对中分子物质抑制红细胞ATP酶活性作用的影响[J].咸宁医学院学报,2001,15(4):254-255.
[239]李晓林,陆艳娟,李晓梅,等.人参二醇皂甙对内毒素诱导休克大鼠模型肺组织超氧化物歧化酶和丙二醛水平的影响[J].中国临床康复,2006,10(15):123-125.
[240]张亚玉,宋晓霞,孙海,等.栽培人参、西洋参不同年限土壤酸碱度和酶活性的变化[J].特产研究,2008(4):34-36.
[241]张岩,金凤燮,鱼红闪.GS0202菌产人参皂苷糖苷酶的酶性质研究[J].食品与发酵工业,2009,35(4):16-19.
[242]吴梧桐.生物化学(第5版)[M].北京:人民卫生出版社,2004:139.
[243]Markert C L, Moller F. Multiple forms of enzymes: tissue, ontogenetic, and species specific patterns[J]. Proc Natl Acad Sci U S ,1959, 45(5):753–763.
[244]胡能书,万国贤.同工酶技术及应用[M].长沙:湖南科技出版社,1985:2-17.
[245]高昆.同工酶技术在植物生态学研究中的应用[J].山西大同大学学报(自然科学版) ,2009,25(2):62-65.
[246]李志忠,吴婷婷.同工酶技术及其应用研究[J].甘肃高师学报(自然科学版),1999,4(2):46-52.
[247]陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2002:239-245.
[248]萧浪涛,王三根.植物生理学实验技术[M].北京:中国农业出版社,2005:82-83.
[249]陈建勋,王晓峰.植物生理学实验指导(第2版)[M].广州:华南理工大学出版社,2002:72-73.
[250]张志良,吴光耀.植物生物化学技术和方法[M].北京:农业出版社,1986:40-42.
[251]张志良,瞿伟菁.植物生理学实验指导(第3版)[M].北京:高等教育出版社,2003: 121-122
[252]彭志英,蒋黎.紫外速率直接法测定过氧化氢酶活性[J].华西医学,1995,10(1):4-7.
[253]杨兰芳,庞静,彭小兰.紫外分光光度法测定植物过氧化物酶活力[J].现代农业科技,2009(20):364-362.
[254] GB/T5009?171-2003.保健食品中超氧化物歧化酶(SOD)活性的测定[S].
[255]俞建英,蒋宇,王善利.生物化学实验技术[M].化学工业出版社,2005:266-269.
[256]张志良,瞿伟菁.植物生理学实验指导(第3版)[M].北京:高等教育出版社, 2002:135-136.
[257]翁霞,辛广,孟玲.小麦酯酶的纯化研究[J].鞍山师范学院学报,2008,10(4):31-34.
[258]王談,钱士匀.生物化学和临床生物化学检验[M].北京:清华大学出版社,2005: 152-155.
[259]余叔文,汤章城.植物生理学与分子生物学(第2版)[M].北京:科学出版社,1998:366 - 389.
[260]蒋明义.水分胁迫下植物体内的OH产生与细胞的氧化损害[J].植物学报,1999,41 (3):229 - 234.
[261]辛树权,赵骥民,何正飚,等.促生菌对黄瓜幼苗过氧化物酶活力的影响[J].长春师范学院学报(自然科学版),2008,27(1):59-62.
[262]张健,刘美艳.土人参的抗氧化成分分析[J].江苏农业科学,2005(1):109-110.
[263]杨淑慎,高俊凤.活性氧、自由基与植物的衰老[J].西北植物学报,2001,21 (2):215– 220
[264]张志娥,石思信,肖建平,等.人参种子后熟期间过氧化物酶同工酶的变化[J].特产研究,1996(1):3-5.
[265]Bonfill M, CusidóR M, Palazón J, et al. Relationship between peroxidase activity andorganogenesis in Panax ginseng calluses[J]. Plant Cell, Tissue and Organ Culture, 2003,73(1):37-41.
[266]胡卓逸,史建勋,丁家宜.人参细胞培养物中过氧化氢酶活力的测定[J].中草药, 1997,28(6):335-337.
[239]李晓林,陆艳娟,李晓梅,等.人参二醇皂甙对内毒素诱导休克大鼠模型肺组织超氧化物歧化酶和丙二醛水平的影响[J].中国临床康复,2006,10(15):123-125.
[267]汪少云,叶秀云,饶平凡.绿豆中苹果酸脱氢酶的性质及生物进化的研究[J].中国食品学报,2006,6(1):262-266.
[268]李洪山,张晓岚,周培之.旱生植物梭梭苹果酸脱氢酶的纯化及其性质的研究[J].新疆大学学报(自然科学版),1994,11(1):72-76.
[269]王以柔,李平,刘鸿先,等.低温对不同耐寒力的黄瓜幼苗子叶各细胞器中NAD+—苹果酸脱氢酶的影响[J].植物生理学报,1985,11(2):147-154.
[270]刘珠耀,范惠琴,王海庭.番茄苹果脱氢酶同工酶分析[J].植物生理通讯,1983,(2):23-26.
[271]刘琳,肖光辉,肖晓玲.酯酶和苹果酸脱氢酶同工酶分析用于香菇原种的鉴定[J].湖南农业科学,2000(3):51-53.
[272]马国龙,孙伟,常洪,等.洼地绵羊苹果酸脱氢酶同工酶研究[J].草食家畜,2006(3):5-7.
[273]丛辉,王惠民,王跃国,等.毛细管电泳法分离血清中乳酸脱氢酶同工酶[J].中华检验医学杂志,2005,28(11):1143-1145.
[274]宋鉴清,邹大伟,吕宁,等.乳酸脱氢酶(LDH)及其同工酶对良、恶性浆膜腔积液的鉴别诊断价值[J].山东医药,2009,49(3):68-69.
[275]Pecina, P, Houstkova, H, Hansikova, H, et al. Genetic Defects of Cytocrhome c Oxidase Assembly[J]. Physiol Res, 2004,53 (Suppl. 1): S213-S223.
[276]郭晓霞,郑哲民,于广志.酯酶同工酶多态性及其在昆虫分类学中的应用价值[J].昆虫知识,2000,37(6):371-374.
[277]王建,郑军,徐志国.单味人参、定志小丸等对小鼠组织胆碱酯酶活力的影响[J].陕西中医学院学报,1994,17(4):37-39.
[278]吴应文.大麦芽中酸性磷酸酯酶的分离[J].甘肃科学学报,1991,3(4):69-71.
[279]陈禅友,万茜.长豇豆种子的酸性磷酸(酯)酶活性分析及其应用[J].江汉大学学报,1999,16(6):18-19.
[280]杨立红,孙振兴,王晓洁,等.刺参酸性磷酸酯酶的分离纯化及部分性质研究[J].食品科学,2008,29(10):440-443.
[281]Yamasaki K H, Yokoyama H, Miyano K Y, et al. Purification and Characterization ofβ-Amylase from Ginseng[J]. Chem Pharm Bull,1989,37(4):973-978.
[282]赵可伟,司徒儒如,巫剑雄,等.血清中葡萄糖6-磷酸异构酶在类风湿关节炎中的诊断价值[J].热带医学杂志,2008,8(8):813-815.
[283]李慧颖,姜国平,吕英春,等.葡萄糖6-磷酸异构酶在类风湿关节炎中的诊断意义[J].吉林医学,2009,30(11):961-963.
[284]刘胜群.人参规范化生产操作技术(GAP)[D].长春:吉林农业大学, 2003,1-6.
[285]姜先刚,赵雨,唐任能,等.鲜人参药材水提取物的高效凝胶过滤色谱指纹图谱研究[J].药物分析,2008, 28(8):1222-1225.
[286]姜先刚,赵雨,张巍,等.人参水溶性蛋白SDS-聚丙烯酰胺凝胶电泳指纹图谱研究[J].药物分析,2008, 28(6):873-876.
[287]Soldati F, Tanaka O. pnnax ginseng: relation between age of Plant and content of gensenosides[J]. Planta Medica, 1984, 51(4):351-352.
[288]王世民.人参不同部位的皂甙组成及含量[J].国外医药﹒植物药分册,1981(5): 43.
[289]吴冬梅,屈海云,卢菲,等.端视电感耦合等离子体原子发射光谱法测定人参不同部位中的微量元素[J].分析科学学报,2007(5):563-566.
[290]王莉.人参与西洋参的比较鉴别[J].海峡药学,2007,19(4):58-59.
[291]陈燕.鲜人参、生晒参和红参的比较研究[J].海峡药学,2006,18(4):137-139.
[292]印敏芳.山参质量的经验鉴别[J].中国药业,2006,15 (9):56-57.
[293]楼余淦,凌明,程存归,等.生晒山参、西洋参、红参和生晒参的FTIR图谱研究[J].中草药,2003,34(7):665-667.
[294]柳良燕.人参、西洋参功效比较及用法研究[J].实用医技杂志,2008,15(34):41-43.
[295]姜海平,窦德强,荆淑琴,等.林下山参的人参皂苷含量分析和指纹图谱研究[J].中国现代中药,2008,10(4):12-15.
[296] Petros A J, Turner S C, Nunn A J. Cost implications of using inhaled nitric oxide compared with epoprostenol for pulmonary hypertension[J]. J Pharm Technol, 1995,11 (4):163-166.
[297]Hsieh M T, Peng W H, Wu C R, et al. The ameliorating effects of the cognitive- enhancing Chinese herbs on scopolamine-induced amnesia in rats[J]. Phytother Res, 2000, 14(5):375- 377.
[298]Wesnes K A, Ward T, McGinty A, et al. The memory enhancing effects of a Ginkgo biloba / Panax ginseng combination in healthy middleaged volunteers[J]. Psychopharmacology, 2000,152 (4):353-361.
[299]Gold J L, Laxer D A, Dergal J M, et al. Herbal drug therapy interactions: a focus on dementia[J]. Curr Opin Clin Nutr Metab Care, 2001, 4 (1): 29-34.
[300]Yoshikawa T, Asada Y, Furuya T. Continuous production of glycosides by a bioreactor using ginseng hairy root culture[J]. Appl Microbiolo Biotechnol, 1993, 39(4-5):460-464.
[301]Asada Y, Saito H, Yoshikawa T, et al. Biotransformation of 18 beta- glycyrrhet inic acid by ginseng hairy root culture[J]. phytochemistry, 1993,34(4):1049-1052.
[302]Gennady P M, Larisa I F, Yuri N Z, et al. Primary structures of two ribonucleases from ginseng calluses: New members of the PR-10 family of intracellular pathogenesis-related plant proteins. FEBS Lett, 1997,407(2):207-210.
[303]Abe1 F, Miura T, Nagahama1 T, et al. Isolation of a highly copper-tolerant yeast, Cryptococcus sp., from the Japan Trench and the induction of superoxide dismutase activity by Cu2+[J]. Biotechnology Lett, 2001,23(24):2027-2034.
[304]Bala K, Tripathy B C, Sharma D. Neuroprotective and anti-ageing effects of curcumin in aged rat brain regions[J]. Biogerontology, 2006,7(2):81-89.
[305]Kobayashi Y, Okahata S, Sakano T, et al. Superoxide dismutase activity of Tlym phocytes and non-T-lymphocytes[J]. FEBS Lett, 1979,98(2):391-393.
[306]Qu Y, Zhang H, Zhao S, et al. The effect on radioresistance of manganese superoxide dismutase in nasopharyngeal carcinoma[J]. Oncol Rep, 2010,23(4):1005-1011.
[307]Dees A, Zahl A, Puchta R, et al. Ivanovic′-Burmazovic′,I., Water exchange on seven-coordinate Mn(II) complexes with macrocyclic pentadentate ligands: insight in the mechanism of Mn(II) SOD mimetics[J]. Inorg Chem, 2007,46(7):2459-2470.
[308]许雅娟,赵艳景,胡虹.邻苯三酚自氧化法测定超氧化物歧化酶活性的研究[J].西南民族大学学报(自然科学版),2006,32(6):1207-1212.
[309]罗广华,王爱国.植物SOD的凝胶电泳及活性的显示[J].植物生理学通讯,1983 (6):44-45.
[310]郭尧军.蛋白质电泳实验技术(第1版)[M].北京:科学出版社,1993:54-56.
[311]Salin M L, Bridges S M. Isolation and Characterization of an Ironcontaining Superxide Dismutase from an Eucaryote Brassica Campestris[J]. Arch: J Biochem Biophys, 1980, 201(2):369-374.
[312]Kwiatowski J, Safianowska A, Kaniuga Z. Isolation and Characterization of an Ironcontaining Superxide Dismutase from Tomato Leaves[J]. Lycoersicon scalentum: Eur J Biochem, 1985,146(2):459-466.
[313]邹国林,罗时文,裘名宜.小白菜线粒体锰超氧化物歧化酶的纯化和性质研究[J].生物化学与生物物理学报,1992,24(2):183-183..
[314]McCord J M, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein)[J]. J Biol Chem, 1969,244(22):6049-6055.
[315]方允中,李文杰.自由基与酶基础理论及其在生物学医学中的应用[M].北京:科学出版社,1989:147-162.
[316]张彩莹.不同来源的超氧化物歧化酶部分理化性质比较研究[J].安徽农业科学,2007,35(15):4426-4429.
[317]王岚.超氧化物歧化酶的研究及应用概况[J].武汉工业学院学报.2002(1):36-40.
[318]Maral J, Puget K, Michelson A M. Comparative study of superoxide dismutase, catalase and glutathione peroxidase levels in erythrocytes of different animals[J]. Biochem Biophys Res Commun, 1977,77(4):1525-1535.
[319]Kobayashi Y, Okahata S, Sakano T, et al. Superoxide dismutase activity of T lymphocytes and non-T lymphocytes[J]. FEBS Lett, 1979,98(2):391-393.
[320]Cone R, Hasan S K, Lown J W, et al. The mechanism of the degradation of DNA by streptonigrin [J]. Can J Biochem, 1976,54(3):219-223.
[321]王琦,徐新莲,贾维峰,等.我国SOD水果生产现状与发展对策(续)[J].中国果业信息,2005(8):9-11.
[322]Manning M C, Patel K, Borchardt R T. Stability of protein pharmaceuticals[J]. Pharm Res, 1989,11(6):903-918.
[323]Arakawa T, Prest relsik S J, Kenney W C, et al. Factor affecting short - term and long– term stabilities of proteins[J]. Adv Drug Deli Rev, 2001,46(1-3):307-326.
[324]Fagain C O. Understanding and increasing protein stability[J]. Bioch Biophy Acta,1995, 1252(6):1-14.
[325] Chi E Y, Krishnan S, Randolph T W, et al. Physical stability of proteins in aqueous solution:mechanism and driving forces in nonnative protein aggregation[J]. Pharm Res, 2003,20(9):1325-1336.
[326]曲春香,沈颂东,王雪蜂,等.用考马斯亮蓝测定植物粗提液中可溶性蛋白质含量方法的研究[J].苏州大学学报(自然科学版),2006,22(2):82.
[327]施东魁,胡春梅.膜分离技术及其在医药生产和研究中的应用[J].中国中药杂志,2006,31(15):1221-1224.
[328]Wang H X, Ng T B. A ribonuclease from Chinese ginseng (Panax ginseng) flowers[J]. Protein Expres Purif, 2004,33(2):195-199.
[329]李红艳.人参水溶性蛋白及鹿茸胶原蛋白的活性研究[D].长春:长春中医药大学,2007.
[330]边兴艳,尹学念.MTT比色法及其应用[J].国外医学?临床生物化学与检验学分册, 1998,19(2):83-85.
[331]陈哲生.噻唑蓝比色法检测细胞的活性[J].上海医学检验杂志,1994,9(4):205-206.
[332]陈哲生.噻唑蓝比色法用来研究人类淋巴细胞和小鼠巨噬细胞的活性[J].上海免疫学杂志,1992,12(5):54-56.
[333]刘郁,刘连新.人参功效再认识[J].时珍国医国药,2006,17(2):289.
[334]蒋景华.人参的药理作用和临床应用[J].现代中西医结合杂志,2004,13(7):956-957.
[335]许见春.人参在实际应用中的药理作用[J].时珍国医国药,2006,17(4):578.
[336]王卫霞,王谦.人参皂苷的免疫调节作用极其应用[J].中华中医药杂志,2005,20 (4):234-236.
[337]梅少林,袁红艳,常雅萍,等.人参皂苷Rg3体外抗HSV-1活性与免疫调节效应[J].吉林大学学报(医学版),2006,32(6):1019-1022.
[338]褚秀玲,苏建青,韦旭斌.人参皂苷免疫调节和抗病毒作用研究进展[J].中兽医医药杂志,2008(5):20-23.
[339]郭志廷.经分子修饰的人参皂苷的免疫调节作用及机理研究[D].吉林大学硕士学位论文,2006.
[340]张晓芬,任伯成.人参抗运动疲劳作用的实验研究[J].中医药学报,1995(6):29.
[341]冯毅翀,潘华山,赵自明,等.运动性疲劳大鼠中枢神经递质改变及人参皂甙Rb1和Re1抗疲劳的实验研究[J].湖北中医杂志,2009,31(3):5-7.
[342]赵文莉,张馨,王伟,等.人参茎叶总皂甙抗疲劳作用的体内实验研究[J].卫生研究, 2009,38(2):184-187.
[343]王斌,张声华,李晓莉,等.人参总皂甙的耐缺氧效应机理研究[J].食品科学,2002,23(8):270-272.
[344]姜正林,吴新明,金淑仪,等.人参与丹参的抗脑缺氧作用.中国应用生理学杂志,2000, 16 (3):201.
[345]杨世杰,吴涛,周鸣,等.人参茎叶总甙耐窒息性缺氧的实验研究[J].吉林大学学报(医学版),1992,18(2):118-120.
[346]冯立明,番华珍,李慧云,等.人参的抗氧化作用[J].中西医结合杂志,1987,7(5):288-290,262.
[347]张纯武,滕红林,柳献云,等.人参皂甙对强直性脊柱炎镇痛和抗炎作用的实验研究[J].浙江中医杂志,2005,40(9):402-403.
[348]刘振明,吕莉莉,张若英,等.人参对吗啡镇痛作用的影响[J].中药药理与临床,1996,23(6):23-24.
[349]邬蓉,高守红,辛海量.人参抗辐射损伤研究概况[J].药学实践杂志,2007,25(6):364-366,392.
[350]姜建华,张学进,王育丽.人参皂苷对60Co照射小鼠不良反应的影响[J].浙江中西医结合杂志,2005,15(11):682-683.
[351]Ivanova T, Han Y, Son H J, et a1. Antimutagenic effect of polysaccharide ginsan extracted from Panax ginseng [J]. Food Chem Toxieol, 2006, 44(4):517-521.
[352]金光辉,刘树铮,刘扬,等.人参组甙对紫外线辐射致皮肤角质形成细胞损伤的保护作用[J].中华放射医学与防护杂志,2002,22(6):420-421.
[353]刘丽波,孙晓玲,张海英,等.人参三醇组苷对小鼠骨髓细胞染色体辐射的防护作用[J].吉林大学学报,2002,28(2):l38-140.
[354]李校坤,林灼锋,黄亚东,等.人参三醇组甙抗辐射作用的研究[J].中药材,2002,25(11):805-808.
[355]刘丽波,李玲,刘鹏来,等.人参皂苷对雄性小鼠骨髓细胞染色体的辐射防护作用.白求恩医科大学学报,1997,4 (23) :348-350.
[356]GPT2-1.中药、天然药物急性毒性研究技术指导原则[Z].2005.
[357]中华人民共和国卫生部.保健食品检验与评价技术规范[Z].2003.
[358]苏丙凡,张智慧,商亚珍.灯盏花素抑制角叉菜胶诱导小鼠血栓形成[J].承德医学院学报,2009,26(1):8-9.
[359]辛晓明,王大伟,王远丽,等.杜仲多糖镇痛作用研究[J].现代中西医结合杂志, 2009, 18(5):487-488.
[360]舒畅,张延英,蔺兴遥,等.红花凝胶剂抗炎镇痛作用的实验研究[J].中国实验方剂学杂志,2009,15(1):72-73.
[361]袁伯俊,廖明阳,李波.药物毒理学实验方法与技术[M].北京:化学工业出版社(生物·医药出版分社),2007:200-201,204.
[362]李晓波,金鑫,李妍,等.植物类中药活性蛋白成分研究进展[J].中草药,2004,35(6) 706-708.
[363]付秀花.中草药组方对小鼠免疫功能的影响[D].南京农业大学硕士学位论文,2003.
[364]吴红星,杨剑婷.抗辐射剂的研究现状[J].放射免疫学杂志,2005,18(1):58-60.
[365]崔凤梅,聂继华,李建祥,等.黑风蜂胶抗辐射损伤的实验研究[J].工业卫生与职业病,2006,32(5):283-285.
[366]邓砚,覃丽佳,谭钧,等.茉莉花提取液对老龄小鼠的抗氧化作用[J].临床研究,2009,6(5):17-18.
[367]郦章安,吴春福.现代老年药学[M].北京:中国医药科技出版社,200:430-441.
[368]Giardino R, Giavaresi G, Fini M, et al. The role of different chemical modifications of superoxide dismu-tase in preventing a prolonged muscular ischemia/reperfusion injury[J]. Artif Cell Blood Sub, 2002,30(3):189-198.
[369]杨江涛,杨娟,杨江冰.刺梨多糖对衰老小鼠体内抗氧化能力的影响[J].营养学报, 2008, 30(4):407-409.
[370]朱伟,孙红光,朱迅.黄芩苷元对炎症反应的影响[J].中国药理学通报,2009,25(2):194- 197.