地锦草总黄酮提取工艺及部分药理学研究
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摘要
地锦草(Euphorbia humifusa)是一种常见中草药,生于原野、荒地、路旁、田间,分布于全国各地。众多学者对其化学成分、药理作用、临床应用做了研究,但对其黄酮类提取物的研究尚浅。本文以地锦草为研究对象,确立了地锦草总黄酮含量测定方法,建立了提取和分离纯化的工艺路线;并对地锦草黄酮抑菌作用、免疫药理作用进行了研究,分别从整体水平、细胞水平和分子水平上探讨其免疫作用机制。研究结果如下:
1、运用正交试验优化超声破碎地锦草细胞的最佳条件,采用超声-加热回流-大孔树脂吸附的工艺流程提取分离纯化地锦草总黄酮,紫外分光光度法在波长510nm处测吸光度测地锦草总黄酮含量。试验结果表明:超声破碎的最佳条件为以料液比为1∶20,70%的乙醇为提取溶剂,60℃超声提取35min。影响超声提取效果大小的因素依次为提取时间＞溶剂浓度＞料液比＞提取温度。100g地锦草干燥全草提取纯化得到3.679g黄褐色粉末,经多种化学试剂鉴定,均成明显的黄酮反应。在3.679g地锦草黄酮粉末中,含纯黄酮3.131g,总得率为3.131%。
2、采用纸片法测定地锦草总黄酮体外抗菌效果,并采用试管倍比稀释法测定其最低抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果表明:地锦草总黄酮对巴氏杆菌,沙门氏菌的体外抑菌作用较强,10mg/ml的总黄酮提取液对巴氏杆菌,沙门氏菌的抑菌直径分别达到22.0±1.0(mm),26.0±2.0(mm),随着黄酮浓度的增加,其抑菌作用也增强。本实验测得地锦草黄酮对巴氏杆菌,沙门氏菌的MIC分别为1.25mg/ml,0.625mg/ml,沙门氏菌的MBC浓度为1.25mg/ml。地锦草总黄酮提取液对金黄色葡萄球菌,大肠杆菌和链球菌也有一定的抑制作用。
3、采用常规免疫药理学试验技术将地锦草总黄酮的3个剂量组与对照组进行比较,结果显示:脾指数、碳廓清指数(K)、吞噬指数(a)、足趾增厚值均显著高于空白组,说明一定剂量的地锦草黄酮具有提高免疫作用的能力。
4、采用RT-PCR测定小鼠脾组织中IL-2、IL-12、IFN-γ、TNF-αmRNA的表达水平,试验结果表明:①H组和M组IL-2 mRNA表达水平均升高,与C组相比,H组和M组极显著差异(P＜0.01);②与对照组相比,三个剂量的地锦草黄酮对IL-12 mRNA表达水平稍有提高,但差异不是很显著(P＞0.05);③TNF-γmRNA表达水平均有提高,特别是H剂量组(P＜0.01);④与对照组相比,各个剂量的地锦草黄酮对TNF-αmRNA表达水平均有升高,且差异显著(P＜0.01)。说明地锦草黄酮三个剂量组均能提高IL-2、IL-12、IFN-γ、TNF-αmRNA的表达水平,从而提高机体的免疫功能与抗病能力。
综上所述,超声-加热回流-大孔树脂吸附提取分离纯化地锦草总黄酮的方法操作简单,重复性好,提取的总黄酮含量高,纯度好。研究表明,地锦草黄酮具有体外抑菌活性;在在一定剂量范围内能有效提高小鼠的免疫功能及IL-2、IL-12、IFN-γ和TNF-αmRNA表达。
Euphorbia humifusa was a common Chinese herbs which was distributed in the wildness,wasteland,roadsides,fields of almost every province of China.Many scholars researched in the chemical composition,pharmacological effects,clinical application of Euphorbia humifusa,but less involved in flavonoid extraction.In this paper,the author conducted a study to establish the method for determination of Euphorbia humifusa total flavonoids,and established the process line of extraction, separation and purification;furthermore,did some research in bacteriostasis,immune pharmacological effect of Euphorbia humifusa flavonoids,respectively,from the overall level,cell level and molecular level to explore the role in the immune mechanism.The results were as follows:
1.To optimize the ultrasonic conditions with orthogonal test,the researcher extratced, separated and purified the total flavonoids with the method of Ultrasonication-Heating reflux-Macroreticular resin adsorbing.The UV-spectrophotometry was adopted with the detection wavelength at 510nm to determinate the content of Euphorbia humifusa total flavonoids.The experimental results showed that:the optimal ultrasonic conditions was solid-liquid ratio as 1:20,ethanol concentration as 70%,the temperature as 60℃,time as 35min.The factors which impact on the efficiency of ultrasonic extraction were as follows by the size of the factors:extraction time＞solvent concentration＞solid-liquid ratio＞temperature.3.679g brown powder was purified from the 100g whole dry Euphorbia humifusa,identified by a variety of chemical reagents,which showed clearly flavone reaction.3.679g extraction powder contained 3.131 g pure flavonoids,which yeild rate was 3.131%.
2.The researcher tested the bacteriostasis effect of Euphorbia humifusa total flavonoids with paper disk method and tested the minimum inhibitory concentration (MIC) and minimum bactericidal concentration(MBC) with double dilution method. The experimental results showed that:The antimicrobial effect of 10mg/ml total flavonoids in vitro against Pasteurella and Salmonella was strong,with the diameter of inhibition zone as 22.0±1.0(mm) and 26.0±2.0(mm),respectively.The total flavonoids MIC was as follows:0.625mg/ml to Pasteurella,1.25mg/ml to Salmonella and MBC was 1.25mg/ml to Salmonella.Euphorbia humifusa total flavonoids extraction also had bacteriostasis effect on Staphylococcus aureus,E.coli and Streptococcus suis to a certain degree.
3.The researcher compared three dose-groups to the control group with the routine immunological pharmacology experimental method.The results showed:mice spleen index,carbon clearance index(K),phagocytic index(a),toes thickening value were significantly higher than the control group,indicating that a certain dose of Euphorbia humifusa flavonoids had the ability to enhance immune function.
4.Using RT-PCR way to test the IL-2,IL-12,IFN-γand TNF-αmRNA expression level in mice spleen tissue,we could know that:①The IL-2 mRNA expression level increased in H group and M group,difference was significant compared with C group(P＜0.01);②The IL-12 mRNA expression level slightly increased in three dose-groups,however,had not obvious difference compared with C group(P＞0.05);③The IFN-γmRNA expression level increased in in three dose-groups,especially H-dose group(P＜0.01);④Compared with C group,The TNF-αmRNA expression level increased in three dose-groups,and significantly different(P＜0.01).It was indicated that Euphorbia humifusa flavonoids were able to enhance the IL-2,IL-12, IFN-γand TNF-αmRNA expression levels,thereby enhancing the immune function and disease resistance.
From the experiment results,we could know that the Ultrasonication-Heating reflux- Macroreticular resin adsorbing method was simple,stable and the total flavonoids extraction was pure and fruitful.Euphorbia humifusa total flavonoids had the bacteriostasis effect in vitro and could improve the immune function of mice and could increased IL-2,IL-12,IFN-γand TNF-αmRNA expression level to certain dose degree.
1、运用正交试验优化超声破碎地锦草细胞的最佳条件,采用超声-加热回流-大孔树脂吸附的工艺流程提取分离纯化地锦草总黄酮,紫外分光光度法在波长510nm处测吸光度测地锦草总黄酮含量。试验结果表明:超声破碎的最佳条件为以料液比为1∶20,70%的乙醇为提取溶剂,60℃超声提取35min。影响超声提取效果大小的因素依次为提取时间＞溶剂浓度＞料液比＞提取温度。100g地锦草干燥全草提取纯化得到3.679g黄褐色粉末,经多种化学试剂鉴定,均成明显的黄酮反应。在3.679g地锦草黄酮粉末中,含纯黄酮3.131g,总得率为3.131%。
2、采用纸片法测定地锦草总黄酮体外抗菌效果,并采用试管倍比稀释法测定其最低抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果表明:地锦草总黄酮对巴氏杆菌,沙门氏菌的体外抑菌作用较强,10mg/ml的总黄酮提取液对巴氏杆菌,沙门氏菌的抑菌直径分别达到22.0±1.0(mm),26.0±2.0(mm),随着黄酮浓度的增加,其抑菌作用也增强。本实验测得地锦草黄酮对巴氏杆菌,沙门氏菌的MIC分别为1.25mg/ml,0.625mg/ml,沙门氏菌的MBC浓度为1.25mg/ml。地锦草总黄酮提取液对金黄色葡萄球菌,大肠杆菌和链球菌也有一定的抑制作用。
3、采用常规免疫药理学试验技术将地锦草总黄酮的3个剂量组与对照组进行比较,结果显示:脾指数、碳廓清指数(K)、吞噬指数(a)、足趾增厚值均显著高于空白组,说明一定剂量的地锦草黄酮具有提高免疫作用的能力。
4、采用RT-PCR测定小鼠脾组织中IL-2、IL-12、IFN-γ、TNF-αmRNA的表达水平,试验结果表明:①H组和M组IL-2 mRNA表达水平均升高,与C组相比,H组和M组极显著差异(P＜0.01);②与对照组相比,三个剂量的地锦草黄酮对IL-12 mRNA表达水平稍有提高,但差异不是很显著(P＞0.05);③TNF-γmRNA表达水平均有提高,特别是H剂量组(P＜0.01);④与对照组相比,各个剂量的地锦草黄酮对TNF-αmRNA表达水平均有升高,且差异显著(P＜0.01)。说明地锦草黄酮三个剂量组均能提高IL-2、IL-12、IFN-γ、TNF-αmRNA的表达水平,从而提高机体的免疫功能与抗病能力。
综上所述,超声-加热回流-大孔树脂吸附提取分离纯化地锦草总黄酮的方法操作简单,重复性好,提取的总黄酮含量高,纯度好。研究表明,地锦草黄酮具有体外抑菌活性;在在一定剂量范围内能有效提高小鼠的免疫功能及IL-2、IL-12、IFN-γ和TNF-αmRNA表达。
Euphorbia humifusa was a common Chinese herbs which was distributed in the wildness,wasteland,roadsides,fields of almost every province of China.Many scholars researched in the chemical composition,pharmacological effects,clinical application of Euphorbia humifusa,but less involved in flavonoid extraction.In this paper,the author conducted a study to establish the method for determination of Euphorbia humifusa total flavonoids,and established the process line of extraction, separation and purification;furthermore,did some research in bacteriostasis,immune pharmacological effect of Euphorbia humifusa flavonoids,respectively,from the overall level,cell level and molecular level to explore the role in the immune mechanism.The results were as follows:
1.To optimize the ultrasonic conditions with orthogonal test,the researcher extratced, separated and purified the total flavonoids with the method of Ultrasonication-Heating reflux-Macroreticular resin adsorbing.The UV-spectrophotometry was adopted with the detection wavelength at 510nm to determinate the content of Euphorbia humifusa total flavonoids.The experimental results showed that:the optimal ultrasonic conditions was solid-liquid ratio as 1:20,ethanol concentration as 70%,the temperature as 60℃,time as 35min.The factors which impact on the efficiency of ultrasonic extraction were as follows by the size of the factors:extraction time＞solvent concentration＞solid-liquid ratio＞temperature.3.679g brown powder was purified from the 100g whole dry Euphorbia humifusa,identified by a variety of chemical reagents,which showed clearly flavone reaction.3.679g extraction powder contained 3.131 g pure flavonoids,which yeild rate was 3.131%.
2.The researcher tested the bacteriostasis effect of Euphorbia humifusa total flavonoids with paper disk method and tested the minimum inhibitory concentration (MIC) and minimum bactericidal concentration(MBC) with double dilution method. The experimental results showed that:The antimicrobial effect of 10mg/ml total flavonoids in vitro against Pasteurella and Salmonella was strong,with the diameter of inhibition zone as 22.0±1.0(mm) and 26.0±2.0(mm),respectively.The total flavonoids MIC was as follows:0.625mg/ml to Pasteurella,1.25mg/ml to Salmonella and MBC was 1.25mg/ml to Salmonella.Euphorbia humifusa total flavonoids extraction also had bacteriostasis effect on Staphylococcus aureus,E.coli and Streptococcus suis to a certain degree.
3.The researcher compared three dose-groups to the control group with the routine immunological pharmacology experimental method.The results showed:mice spleen index,carbon clearance index(K),phagocytic index(a),toes thickening value were significantly higher than the control group,indicating that a certain dose of Euphorbia humifusa flavonoids had the ability to enhance immune function.
4.Using RT-PCR way to test the IL-2,IL-12,IFN-γand TNF-αmRNA expression level in mice spleen tissue,we could know that:①The IL-2 mRNA expression level increased in H group and M group,difference was significant compared with C group(P＜0.01);②The IL-12 mRNA expression level slightly increased in three dose-groups,however,had not obvious difference compared with C group(P＞0.05);③The IFN-γmRNA expression level increased in in three dose-groups,especially H-dose group(P＜0.01);④Compared with C group,The TNF-αmRNA expression level increased in three dose-groups,and significantly different(P＜0.01).It was indicated that Euphorbia humifusa flavonoids were able to enhance the IL-2,IL-12, IFN-γand TNF-αmRNA expression levels,thereby enhancing the immune function and disease resistance.
From the experiment results,we could know that the Ultrasonication-Heating reflux- Macroreticular resin adsorbing method was simple,stable and the total flavonoids extraction was pure and fruitful.Euphorbia humifusa total flavonoids had the bacteriostasis effect in vitro and could improve the immune function of mice and could increased IL-2,IL-12,IFN-γand TNF-αmRNA expression level to certain dose degree.
引文
[1]Ch p(中国药典)[S].Vol Ⅰ.2005
[2]中国医学科学院药用植物资源开发研究所等.中药杂志(第4册)[M].北京:人民卫生出版社,1998.320.
[3]李时珍(明).本草纲目(校点本)(上册)[M].北京:人民卫生出版社,1982.1395.
[4]江纪武.等.植物药有效成分手册[M].北京人民卫生出版社,1986.942、1097.
[5]朱英.地锦草的研究进展[J],现代中药研究与实践.2003,17(5):62-63
[6]柳润辉,孔令义.地锦草脂溶性成分研究[J].天然产物研究与开发,2005,17(4):437
[7]柳润辉,王汉波,孔令义.地锦草化学成分研究[J].中草药,2001,32(2):107
[8]柳润辉,孔令义.斑地锦的化学成分[J].植物资源与环境学报,2001,10(1):60
[9]Agata I,Hatano T,Nakaya Y,et al.Tannins and related polyhenols of Euphorbiaceous Plants.Ⅷ.Eumaculin A and Eusupinin A.and ac companying polyphenols from Euphorbia maculata L.and E.supina Ra-6n[J].Chem Pharm Bull,1991,39(4):881
[10]Yoshida T,Amakura Y,Liu Yah—Ze,et al.Tanins and related polyphenols of euphorbiaceous plant.Ⅺ.Three new hydrolyzable tanins and a polyphenol glucoside from Euphorbia humifusa[J].Chem Pharm Bull,1994,42(9):1803
[11]Yoshiaki A,Keita K,Tsutoma H,et al.Four new Hydrocyzable Tannins And an Acylated Flavonol Glycoside from Euphorbiamaeulate[J].Canadian Journal of Chemistry,1997,75(6):727
[12]Fei Deng,Na Tang,Jun Xu et al.New dpyrrolidinonoids and glycosides from Euphorbia humifusa[J].Joumal of Asian Natural Products Research,2008,10(6):531
[13]古力娜·达吾提,努尔买买提·艾买提,尤丽都孜·买买提,等.雅丽蔓总黄酮体外抗真菌作用研究[J].中国民族医药杂志,2006,4:44
[14]谭红,刘伟.地锦草总黄酮的休外抑菌试验[J].中国兽医医药杂志,2007(4):5
[15]Shimizu M,Shiota S,Mizushima T,et al.Marked Potentiation of Activity of β-Lactams against Methicillin-Resistant Staphylococus aureus by Corilagin[J].Antimicrobial agents and chemotheraPy,2001,445(11):3198
[16]斯拉甫,阿布都热依木,努尔买买提,等.夏塔热片合用其软膏的免疫调节功能[J].中国免疫学杂志,2000,16(5):269
[17]斯拉甫,哈木拉提,努尔买买提.夏塔热片合用夏塔热软膏的抗真菌消炎作用研究[J]. 中草药,2000,31(7):538
[18]努尔买买提,斯拉甫·艾白,哈木拉提·吾甫尔.夏塔热片合用夏塔热软膏的抗过敏作用研究[J].中药新药与临床药理,2003,14(1):24
[19]邓淑文,哈木拉提.百癣夏塔热对皮肤癣菌超微结构、蛋白及细胞膜麦角甾醇生物合成影像的研究[DB/OL].http//www.epub.cnki.net.2004-12-16/2008-07-21
[20]Maertens JA.History of the development of azole derivatives[J].Clin Microbiol and Infect,2004,10(1):1
[21]古力娜·达吾提,尤丽吐孜,艾则孜·亚森,等.维药地锦草软膏的体外抗真菌及其对豚鼠皮肤真菌感染的治疗作用研究[J].中药药理与临床,2007,23(5):178
[22]曹瑞珍,张国文,薛燕,等.地锦草提取物xCzl,鼠血液过氧化氢酶和谷胱甘肽过氧化物酶活性的影响[J].时珍国医国药,2002,13(12):724
[23]曹瑞珍,魏永春,张国文,等.地锦草总黄酮对D一半乳糖衰老模型小鼠抗氧化作用的研究[J].卫生研究,2007,36(3):387
[24]阿不都热依木,阿不都艾尼,哈木拉提,等.五种维吾尔药的清除羟自由基及抗DNA损伤作用研究[J].中草药,2001,32(3):236
[25]阿不都热依木,斯拉甫,热孜万古丽,等.夏塔热片对多种活性氧自由基的清除作用研究[A].新疆维吾尔自治区药学会.新疆药学(新疆第十二届药学学术年会论文集下册)[C].乌鲁木齐:新疆人民卫生出版社,1999:44.
[26]曹瑞珍,魏永春,强欣,等.地锦草总黄酮预防四氯化碳所致急性肝损伤的实验研究[J].时珍国医国药,2007,18(1):85
[27]董福轮,季蓓等.鸟不宿.地锦草降低血糖的临床研究[J].四川中医.2002.20(4):24-25
[28]董鹏,唐万斌,郭连芳.地锦草的止血作用研究[J].武警医学院学报,1996,5(1):21
[29]万亚坤,周乐,赵海双.中药地锦草的研究与临床应用[J].动物医学进展.2000.21(4):157-158
[30]马朝斌.“复方地锦草汤”治疗慢性特发性血小板减少症35例[J].江苏中医药,2004,25(11):31
[31]罗宏新,陈凯.鼻内镜下射频热凝联合地锦草片治疗难愈性鼻出血[J].实用医学杂志,2007,23(8):1228.
[32]周志群.地锦乌茜汤治疗青春期功能性子宫出血76例[J].中国医学文摘,2002,2(8):974
[33]孙来银.葛根地锦草汤治疗慢性结肠炎58例[J].吉林中医药,2000,5:24
[34]热孜万·吾买尔,依力哈木·买买提.百癣夏塔热片治疗57例寻常性银屑病疗效观察 [J].中国民族医药杂志,2006,4:94
[35]木巴拉克,亚尔买买提.夏塔热治疗疮疖临床观察[A].木哈拉提·吾甫尔,阿不都热依木·卡地尔.维吾尔医药及其他传统医学研究与应用(2003国际维吾尔医药学术会议论文集)[C].乌鲁木齐:新疆人民卫生出版社,2003:41
[36]沈军.百癣夏塔热片等治疗成人丘疹性荨麻疹[J].中国中西医结合皮肤性病学杂志,2007,6(2):94
[37]张贤彬,李彩霞,吴亚卿.黄酮类化合物的研究进展[J].食品与机械,2005,5(21):70-73
[38]肖坤福,廖晓峰.黄酮类化合物研究进展及应用[J].食品研究及开发,2003,4(24):13-15
[39]Antibacterial activity of flavonoids against methicillin resisten slaphy lococcu saureus strains[J].J.Theor.Biol,2000,21:205(2):231-240
[40]Calzada F,Meckes M,Cedillo-rivera R.Antiamoebic and antigiardial activity of plant flavonoids.Planta Med,1999,65:78-80
[41]汪德清,沈文梅,田亚平等.黄芪的三种提取成分对氧自由基作用的影响[J].中国药理学通报,1994,10:129
[42]陈琪,王伯初,唐春红.黄酮类化合物抗氧化性与其构效的关系[J].重庆大学学报,2003,11(26):48-51
[43]Asgary S,Naderi G,Sarrafzadegan N,et al.Anti-oxidant effect of flavonoidson hemoglobin glycosylation.Pharmacta Helv,1999,73:223-226
[44]胡春,丁霄霖.黄酮类化合物在不同氧化体系中的抗氧化作用研究[J].食品与发酵工业,1996,22(3):46-53
[45]Matsuzaki Y,Kurokawa N,Terai S,et al.Cell Death Induced by Baicalein in Human hepatocelluar Carcinoma Cell Lines.Jpn J Can-cer Res,1996,87:170
[46]Makela s,poutanen M,kostian M L,et al.Inhibition of 17 betahydr oxysteroid oxidoreductase by flavonoids in breast and prostate cancer cells.Proc Soc Exp Biol Med,1998,217:310-316
[47]李庆林,陈志武,马传庚.黄蜀葵花总黄酮对心肌损伤的保护作用及其机制[J].中国药理学通报,2001,17(4):4661
[48]周兰兰,岑德意,陈志武,等,黄蜀葵花总黄酮对心脑缺氧损伤的保护作用[J].中国中医基础医学杂志,2000,6(7):221
[49]张雪,丁长河.紫外分光光度法检测山植总黄酮方法的建立[J].食品工业科技.2007,28(7):21-22
[50]张赞彬,李彩侠,吴亚卿.黄酮类化合物的研究进展[J].食品与机械.2005,21(5):70-73
[51]毕丽君.香椿嫩叶中黄酮类化合物的提取[J].浙江林学院学报,2000,17(2):146-149
[52]郭孝武.超声提取与常规提取对部分中药碱类成分提出率的比较[J].世界科学技术,2002,5(4):59-61
[53]陈晓慧,徐雅琴.酶法提取树莓叶黄酮类化合物的研究[J].食品研究与开发,2007,2(28):35-36
[54]郭庆祝,刘玉.纤维素酶法提取天然辣椒素的研究[J].现代食品科技,2007,6(96):45-48
[55]王晓,李林波,马小来.酶法提取山楂中总黄酮的研究[J].工业技术,2002,3(23):37-38
[56]邓启焕,二类超临界流体萃取银杏叶有效成分的实验研究[J].中草药,1999,30(6):419-423
[57]张睿,徐雅琴,时限黄酮类化合物提取工艺研究[J].食品与机械.2003,21-22
[58]张蔚君,朱莉,董小莉,等.大孔吸附树脂在中草药成分提纯中的应用及其影响因素,空军总医院学报.2006,2(22):96-98
[59]卢锦花,胡小玲,岳红,等.大孔吸附树脂对银杏叶黄酮类化合物吸附及解吸附的研究[J].化学研究与应用,2002,14(2):164-167
[60]王满力,陈桐.柱色谱法分离地榆中的鞣质和皂甙[J].色谱,1994,4(12):24
[61]华辉,郭勇.黄酮类化合物药理研究进展.广东药学[J].1999(9)4:9-12
[62]郭雪微,卢挺,王雪里红.天然黄酮类化合物在心脑血管疾病中应用进展[J]中国老年学杂志.2006,26(7):1001-1003
[63]李楠,刘元,侯滨滨.黄酮类化合物的功能特性[J].食品研究与.2005,26(6):39-141
[64]夏文宽.菜芙蓉花总黄酮的提取与抗氧化活性研究[D].江苏大学.2006
[65]买买提江,龚千锋.地锦草的研究概况[J].江西中医学院学报.2008,20(4):78-80
[66]安慧霞,李自建,等.地锦草的研究进展[J].时珍国医国药.2008,19(12):2866-2868
[67]洪雪娥,高荫榆,罗丽萍,等.大孔树脂对薯蔓黄酮吸附分离特性研究.食品科学[J].2006,27(10):423-427
[68]高静,刘培勋,龙伟.中药提取技术的研究进展[J].医药导报,2007,9(26):158-159
[69]刘辉林.中草药化学成分提取新技术[J].牙膏工业,2003,3:39-40
[70]陈水英,孙彩华.大孔吸附树脂在天然药物有效成分分离、富集中的应用[J].中药新药与临床药理,2007,18(16):62-63
[71]刘鄂湖,鄢丹,蔡光明等.大孔吸附树脂技术在中药中应用现存问题分析与探讨[J].中草药,2007,5(38):附3-附5
[72]贝伟剑,彭文烈,罗杰.柿叶黄酮的大孔吸附树脂分离提纯富集[J].中成药,2005,(3):257-261
[73]陈乃富,陈科,张莉等.大孔树脂柱层析法纯化金樱子总黄酮的初步研究[J].中药材,2007,8(30):1013-1015
[74]郑亚杰,张长弓,李晓斌.大孔吸附树脂分离纯化山楂总黄酮的研究[J].华中科技大学学报·医学版,2004,33(2):136-138
[75]房方,张广强,谢新年.用分光光度法进行黄酮类化合物含量测定的研究[J].河南中医药学刊,1997,2(12):17-19
[76]Mostafa K Mesbah,Sherief I Khalifa,Alaa El-Gindy,et al.HPLC determination of 10cerain flavonoids and terpenelactones in selected Ginkgo biloba L.phytopharmaceuticals.Ⅱ Farmaco,2005,60(6-7):583-590
[77]Yit Hong Loon,Jia woei Wong,Siew Ping Yap,et al.Determination of flavonoids from orthosiphon stamineus in plasma using simple HPLC method with Ultra-violet detection,Journal of Chromatography B,2005,816(1-2):161-166
[78]魏永生,王永宁,石玉平等.分光光度法测定总黄酮含量的实验条件研究[J].青海大学学报(自然科学版),2003,3(21):61-63
[79]丁航,梁统,周克元等.高效液相色谱法及紫外可见分光光度法测定侧柏叶中黄酮含量的比较[J].广东医学院学报,2002,6(20):422-423
[80]J Sawai,R Doi,Y Maekawa,et al.Indirect conductimetric assay of Antibacterial activities[J].Journal of Industrial Microbiology And Biotechnology,2002,29(5):296-298.
[81]Osamu Yamamoto,Miyako Komatsu,Jun Sawai,et al.Effect of lattice constant of zinc oxide on antibacterial characteristics[J].Journal of Materials Science:Materials in Medicine,2004,15(8):847-851
[82]谢鹏,张敏红.黄酮类化合物抑菌作用的研究进展[J].中国动物保健.2004.12:35-36
[83]井乐刚,张永忠,田璐.大豆异黄酮抑茵活性的研究[J].生物技术通讯.2004,15(5):476-479
[84]邵留,沈盎绿,郑曙明.斑地锦总黄酮的提取及抑菌作用[J],西南农业大学学报(自然科学版),2005,126(6):903-905
[85]Steven C Gllman and Thomas J.Rogers.Immunopharmacology[M].Fir-St ed.New Jersey:The Telfond press,1989,50-100
[86]Bond J S,Butler P E.Intracellular proteases.Annu Rev Biochem.1987,56:333-64
[87]Berfram G Katzung,ed.Basic and clinical pharmacology[M].Sixthed.East Norwalk:Appleton & lange,1995,895-878
[88]国家中医药管理局.中华本草第四册[M].上海出版社.1999,3576.
[89]Yanhua Lu,Chongwei Zhang,Peter Bucheli,et al.Citrus Flavonoids In Fruitand Traditional Chinese Medicinal Food Ingredients in China[J].Plant Foods for Human Nutrition,2006,61(2):55-63
[90]张雪梅,鞠玉琳,熊焕章.中药制剂的免疫药理学研究[J].中国兽医杂志,2008,6(44):44-45
[91]朱正美,刘辉.简明免疫学技术[M].北京:科学出版社,2002,205-206、220-221.
[92]王伟,白俊杰,劳海华,等.草鱼Mx蛋白基因的克隆与原核表达[J].中国水产科学,2003,10(5):366-368
[93]马建国,于江.槐白皮对机体免疫功能影响的研究[J].山东中医杂志,2002,21(9):557-559
[94]F Cappello.Is CDIa involved in antitumour immune responses during carcinogenesis[J].British Journal of Cancer,2004,90(23):938
[95]张兰玲,孟济明,陈海生,等.点腺过路黄正丁醇提取部分对小鼠免疫反应的影响[J].中药材,2002,25(12):888-890
[96]马晓彤,王庭欣,蒋东升,等.一种人参保健品功效作用的评价[J].卫生毒理学杂志,2003,17(1):61-62
[97]阎莉,郑作文,卫智权.广西藤茶总黄酮对正常小鼠免疫功能的影响[J].中国药物应用与监测,2008,5(4):5-7
[98]李小定,吴谋成,曾晓波,等.灰树花多糖粗品与纯品的抗肿瘤作用及对免疫功能的影响[J].营养学报,2003,25(1):7-9
[99]徐叔云,卞如镰,陈修.药理实验方法学(第三版)[M].北京:人民卫生出版社,2002.
[100]朱立平,陈学清.免疫学常用实验方法[M].北京:人民军医出版社,2000:175-180.
[101]戴伟娟,司端运,苏艾兰,等.无花果多糖对小鼠迟发型超敏反应的影响[J].济宁医学院学报,1999,22(4):26-27
[102]伍章保,沈玉先,魏伟,等.富硒酵母对免疫功能低下小鼠DTH的影响[J].中国药理学通报,1999,12(2):182-184
[103]胡相光,洪净,尹华熙,等.凉血清营颗粒对炎症及迟发型超敏反应的影响[J].中药药理与临床,2004,20(1):22-24
[104]杨保华,陈德宇,肖顺汉.消风胶囊对小鼠迟发型超敏反应的影响及其作用机制研究[J].中国中西医结合皮肤性病学杂志,2003,2(1):13-15
[105]张立煌,姚航平,曹雪涛,等.吡喹酮联合IL-18基因治疗血吸虫感染小鼠对免疫功能的影响[J].中国微生物与免疫学杂志,2001,21(1):87-90
[106]张丽丽,张竞逵,聂宏刚,等.不同浓度细辛提取物体外免疫抑制作用的实验研究[J].中华中西医杂志,2003,4(4):481-482
[107]张晓君,祝晨,胡黎,等.党参多糖对小鼠免疫和造血功能的影响[J].中药新药与临床药理,2003,14(14):174-176
[108]李桂峰,康裕财,孙际佳,等.酵母多糖对赤眼蹲非特异性免疫机能的影响[J].中山大学学报(自然科学版),2003,7(4):55-58
[109]林慧彬,林建强,杭建群,等.山东产四种冤丝子免疫增强作用的比较研究[J].中西医结合学报,2003,1(1):51-53
[110]石学魁,张晓莉,宋宝辉,等.五味消毒饮增强小鼠免疫功能的研究[J].细胞与分子免疫学杂志,2000,16(1):85
[111]沈关心,周汝麟.现代免疫学实验技术[M].武汉:湖北科学技术出版社,1998,255-256.
[112]Claudio Fiocchi,Kenneth R.Youngman,Belinda Yen-Lieberman,etal.Modulation of intestinal immune reactivity by interleukin 2Phenotypic and functional analysis of lymphokine activated kilLer cells from human intestinal mucosa[J].Digestive Diseases and Sciences(Historical Archive),1988,33(10):1305-1315
[113]Ar Neng Lai,Joseph C.K.Leung,Fernand Mac-Moune Lai,et al.Tlymphocyte activation in IgA nephropathy:Serum-soluble interleukin 2 receptor level,interleukin 2production,and interleukin 2 receptor expression by cultured lymphocytes[J].Journal of Clinic Al Immunology(Historical Archive),1989,9(6):485-492
[114]Lucie M.Heinzerling Karsten Feige,Stefan Rieder,Margarete K.Akens,et al.Tumor regression induced by intratumoral injection of DNA coding for human interleukin 12into melanoma metastases in gray horses[J].Journal of Molecular Medicine,2001,78(12):692-702
[115]A.Fayyazi,S.Schweyer,A.Soruri,et al.R.Parwaresch.Tlymphocytes and altered keratinocytes express interferon-γ and interleukin 6 in lichen planus[J].Archives of Dermatological Research,1999,291(9):485-490
[116]M.Yamada,Y.Arao,A.Hatano.S.Ni.Effect of recombinant mouse interferon-β on acute and latent herpes simplex infection in mice[J].Archives of Virology(Historical Archive),1988,99(1-2):101-109
[117]Y.Uchida,K.Ohba,A.Ogawa.T.Muraki.Protein kinase C mediates rumor necrosis factor-αinduced inhibition of obese gene expression and leptin secretion in brown adipocytes[J].Naunyn-Schmiedeberg's Archives of Pharmacology,1999,360(6):691-698
[118]刘清池,梁冰.中药诱生细胞因子研究进展[J].中医杂志,1997,38(12):47
[119]李宏梅、李祥瑞、郭慧君.白细胞介素-2的研究进展及应用[J].山东畜牧兽医,2002(5):38-40
[120]Yasuhiro Kodera,Hayao Nakanishi,Seiji Ito,et al.Quantitative detection of disseminated cancer cells in the greater omentum of gastric carcinoma patients with real-time RT-PCR:a comparison with peritoneal lavage cytology[J].Gastric Cancer,2002,5(2):69-70
[2]中国医学科学院药用植物资源开发研究所等.中药杂志(第4册)[M].北京:人民卫生出版社,1998.320.
[3]李时珍(明).本草纲目(校点本)(上册)[M].北京:人民卫生出版社,1982.1395.
[4]江纪武.等.植物药有效成分手册[M].北京人民卫生出版社,1986.942、1097.
[5]朱英.地锦草的研究进展[J],现代中药研究与实践.2003,17(5):62-63
[6]柳润辉,孔令义.地锦草脂溶性成分研究[J].天然产物研究与开发,2005,17(4):437
[7]柳润辉,王汉波,孔令义.地锦草化学成分研究[J].中草药,2001,32(2):107
[8]柳润辉,孔令义.斑地锦的化学成分[J].植物资源与环境学报,2001,10(1):60
[9]Agata I,Hatano T,Nakaya Y,et al.Tannins and related polyhenols of Euphorbiaceous Plants.Ⅷ.Eumaculin A and Eusupinin A.and ac companying polyphenols from Euphorbia maculata L.and E.supina Ra-6n[J].Chem Pharm Bull,1991,39(4):881
[10]Yoshida T,Amakura Y,Liu Yah—Ze,et al.Tanins and related polyphenols of euphorbiaceous plant.Ⅺ.Three new hydrolyzable tanins and a polyphenol glucoside from Euphorbia humifusa[J].Chem Pharm Bull,1994,42(9):1803
[11]Yoshiaki A,Keita K,Tsutoma H,et al.Four new Hydrocyzable Tannins And an Acylated Flavonol Glycoside from Euphorbiamaeulate[J].Canadian Journal of Chemistry,1997,75(6):727
[12]Fei Deng,Na Tang,Jun Xu et al.New dpyrrolidinonoids and glycosides from Euphorbia humifusa[J].Joumal of Asian Natural Products Research,2008,10(6):531
[13]古力娜·达吾提,努尔买买提·艾买提,尤丽都孜·买买提,等.雅丽蔓总黄酮体外抗真菌作用研究[J].中国民族医药杂志,2006,4:44
[14]谭红,刘伟.地锦草总黄酮的休外抑菌试验[J].中国兽医医药杂志,2007(4):5
[15]Shimizu M,Shiota S,Mizushima T,et al.Marked Potentiation of Activity of β-Lactams against Methicillin-Resistant Staphylococus aureus by Corilagin[J].Antimicrobial agents and chemotheraPy,2001,445(11):3198
[16]斯拉甫,阿布都热依木,努尔买买提,等.夏塔热片合用其软膏的免疫调节功能[J].中国免疫学杂志,2000,16(5):269
[17]斯拉甫,哈木拉提,努尔买买提.夏塔热片合用夏塔热软膏的抗真菌消炎作用研究[J]. 中草药,2000,31(7):538
[18]努尔买买提,斯拉甫·艾白,哈木拉提·吾甫尔.夏塔热片合用夏塔热软膏的抗过敏作用研究[J].中药新药与临床药理,2003,14(1):24
[19]邓淑文,哈木拉提.百癣夏塔热对皮肤癣菌超微结构、蛋白及细胞膜麦角甾醇生物合成影像的研究[DB/OL].http//www.epub.cnki.net.2004-12-16/2008-07-21
[20]Maertens JA.History of the development of azole derivatives[J].Clin Microbiol and Infect,2004,10(1):1
[21]古力娜·达吾提,尤丽吐孜,艾则孜·亚森,等.维药地锦草软膏的体外抗真菌及其对豚鼠皮肤真菌感染的治疗作用研究[J].中药药理与临床,2007,23(5):178
[22]曹瑞珍,张国文,薛燕,等.地锦草提取物xCzl,鼠血液过氧化氢酶和谷胱甘肽过氧化物酶活性的影响[J].时珍国医国药,2002,13(12):724
[23]曹瑞珍,魏永春,张国文,等.地锦草总黄酮对D一半乳糖衰老模型小鼠抗氧化作用的研究[J].卫生研究,2007,36(3):387
[24]阿不都热依木,阿不都艾尼,哈木拉提,等.五种维吾尔药的清除羟自由基及抗DNA损伤作用研究[J].中草药,2001,32(3):236
[25]阿不都热依木,斯拉甫,热孜万古丽,等.夏塔热片对多种活性氧自由基的清除作用研究[A].新疆维吾尔自治区药学会.新疆药学(新疆第十二届药学学术年会论文集下册)[C].乌鲁木齐:新疆人民卫生出版社,1999:44.
[26]曹瑞珍,魏永春,强欣,等.地锦草总黄酮预防四氯化碳所致急性肝损伤的实验研究[J].时珍国医国药,2007,18(1):85
[27]董福轮,季蓓等.鸟不宿.地锦草降低血糖的临床研究[J].四川中医.2002.20(4):24-25
[28]董鹏,唐万斌,郭连芳.地锦草的止血作用研究[J].武警医学院学报,1996,5(1):21
[29]万亚坤,周乐,赵海双.中药地锦草的研究与临床应用[J].动物医学进展.2000.21(4):157-158
[30]马朝斌.“复方地锦草汤”治疗慢性特发性血小板减少症35例[J].江苏中医药,2004,25(11):31
[31]罗宏新,陈凯.鼻内镜下射频热凝联合地锦草片治疗难愈性鼻出血[J].实用医学杂志,2007,23(8):1228.
[32]周志群.地锦乌茜汤治疗青春期功能性子宫出血76例[J].中国医学文摘,2002,2(8):974
[33]孙来银.葛根地锦草汤治疗慢性结肠炎58例[J].吉林中医药,2000,5:24
[34]热孜万·吾买尔,依力哈木·买买提.百癣夏塔热片治疗57例寻常性银屑病疗效观察 [J].中国民族医药杂志,2006,4:94
[35]木巴拉克,亚尔买买提.夏塔热治疗疮疖临床观察[A].木哈拉提·吾甫尔,阿不都热依木·卡地尔.维吾尔医药及其他传统医学研究与应用(2003国际维吾尔医药学术会议论文集)[C].乌鲁木齐:新疆人民卫生出版社,2003:41
[36]沈军.百癣夏塔热片等治疗成人丘疹性荨麻疹[J].中国中西医结合皮肤性病学杂志,2007,6(2):94
[37]张贤彬,李彩霞,吴亚卿.黄酮类化合物的研究进展[J].食品与机械,2005,5(21):70-73
[38]肖坤福,廖晓峰.黄酮类化合物研究进展及应用[J].食品研究及开发,2003,4(24):13-15
[39]Antibacterial activity of flavonoids against methicillin resisten slaphy lococcu saureus strains[J].J.Theor.Biol,2000,21:205(2):231-240
[40]Calzada F,Meckes M,Cedillo-rivera R.Antiamoebic and antigiardial activity of plant flavonoids.Planta Med,1999,65:78-80
[41]汪德清,沈文梅,田亚平等.黄芪的三种提取成分对氧自由基作用的影响[J].中国药理学通报,1994,10:129
[42]陈琪,王伯初,唐春红.黄酮类化合物抗氧化性与其构效的关系[J].重庆大学学报,2003,11(26):48-51
[43]Asgary S,Naderi G,Sarrafzadegan N,et al.Anti-oxidant effect of flavonoidson hemoglobin glycosylation.Pharmacta Helv,1999,73:223-226
[44]胡春,丁霄霖.黄酮类化合物在不同氧化体系中的抗氧化作用研究[J].食品与发酵工业,1996,22(3):46-53
[45]Matsuzaki Y,Kurokawa N,Terai S,et al.Cell Death Induced by Baicalein in Human hepatocelluar Carcinoma Cell Lines.Jpn J Can-cer Res,1996,87:170
[46]Makela s,poutanen M,kostian M L,et al.Inhibition of 17 betahydr oxysteroid oxidoreductase by flavonoids in breast and prostate cancer cells.Proc Soc Exp Biol Med,1998,217:310-316
[47]李庆林,陈志武,马传庚.黄蜀葵花总黄酮对心肌损伤的保护作用及其机制[J].中国药理学通报,2001,17(4):4661
[48]周兰兰,岑德意,陈志武,等,黄蜀葵花总黄酮对心脑缺氧损伤的保护作用[J].中国中医基础医学杂志,2000,6(7):221
[49]张雪,丁长河.紫外分光光度法检测山植总黄酮方法的建立[J].食品工业科技.2007,28(7):21-22
[50]张赞彬,李彩侠,吴亚卿.黄酮类化合物的研究进展[J].食品与机械.2005,21(5):70-73
[51]毕丽君.香椿嫩叶中黄酮类化合物的提取[J].浙江林学院学报,2000,17(2):146-149
[52]郭孝武.超声提取与常规提取对部分中药碱类成分提出率的比较[J].世界科学技术,2002,5(4):59-61
[53]陈晓慧,徐雅琴.酶法提取树莓叶黄酮类化合物的研究[J].食品研究与开发,2007,2(28):35-36
[54]郭庆祝,刘玉.纤维素酶法提取天然辣椒素的研究[J].现代食品科技,2007,6(96):45-48
[55]王晓,李林波,马小来.酶法提取山楂中总黄酮的研究[J].工业技术,2002,3(23):37-38
[56]邓启焕,二类超临界流体萃取银杏叶有效成分的实验研究[J].中草药,1999,30(6):419-423
[57]张睿,徐雅琴,时限黄酮类化合物提取工艺研究[J].食品与机械.2003,21-22
[58]张蔚君,朱莉,董小莉,等.大孔吸附树脂在中草药成分提纯中的应用及其影响因素,空军总医院学报.2006,2(22):96-98
[59]卢锦花,胡小玲,岳红,等.大孔吸附树脂对银杏叶黄酮类化合物吸附及解吸附的研究[J].化学研究与应用,2002,14(2):164-167
[60]王满力,陈桐.柱色谱法分离地榆中的鞣质和皂甙[J].色谱,1994,4(12):24
[61]华辉,郭勇.黄酮类化合物药理研究进展.广东药学[J].1999(9)4:9-12
[62]郭雪微,卢挺,王雪里红.天然黄酮类化合物在心脑血管疾病中应用进展[J]中国老年学杂志.2006,26(7):1001-1003
[63]李楠,刘元,侯滨滨.黄酮类化合物的功能特性[J].食品研究与.2005,26(6):39-141
[64]夏文宽.菜芙蓉花总黄酮的提取与抗氧化活性研究[D].江苏大学.2006
[65]买买提江,龚千锋.地锦草的研究概况[J].江西中医学院学报.2008,20(4):78-80
[66]安慧霞,李自建,等.地锦草的研究进展[J].时珍国医国药.2008,19(12):2866-2868
[67]洪雪娥,高荫榆,罗丽萍,等.大孔树脂对薯蔓黄酮吸附分离特性研究.食品科学[J].2006,27(10):423-427
[68]高静,刘培勋,龙伟.中药提取技术的研究进展[J].医药导报,2007,9(26):158-159
[69]刘辉林.中草药化学成分提取新技术[J].牙膏工业,2003,3:39-40
[70]陈水英,孙彩华.大孔吸附树脂在天然药物有效成分分离、富集中的应用[J].中药新药与临床药理,2007,18(16):62-63
[71]刘鄂湖,鄢丹,蔡光明等.大孔吸附树脂技术在中药中应用现存问题分析与探讨[J].中草药,2007,5(38):附3-附5
[72]贝伟剑,彭文烈,罗杰.柿叶黄酮的大孔吸附树脂分离提纯富集[J].中成药,2005,(3):257-261
[73]陈乃富,陈科,张莉等.大孔树脂柱层析法纯化金樱子总黄酮的初步研究[J].中药材,2007,8(30):1013-1015
[74]郑亚杰,张长弓,李晓斌.大孔吸附树脂分离纯化山楂总黄酮的研究[J].华中科技大学学报·医学版,2004,33(2):136-138
[75]房方,张广强,谢新年.用分光光度法进行黄酮类化合物含量测定的研究[J].河南中医药学刊,1997,2(12):17-19
[76]Mostafa K Mesbah,Sherief I Khalifa,Alaa El-Gindy,et al.HPLC determination of 10cerain flavonoids and terpenelactones in selected Ginkgo biloba L.phytopharmaceuticals.Ⅱ Farmaco,2005,60(6-7):583-590
[77]Yit Hong Loon,Jia woei Wong,Siew Ping Yap,et al.Determination of flavonoids from orthosiphon stamineus in plasma using simple HPLC method with Ultra-violet detection,Journal of Chromatography B,2005,816(1-2):161-166
[78]魏永生,王永宁,石玉平等.分光光度法测定总黄酮含量的实验条件研究[J].青海大学学报(自然科学版),2003,3(21):61-63
[79]丁航,梁统,周克元等.高效液相色谱法及紫外可见分光光度法测定侧柏叶中黄酮含量的比较[J].广东医学院学报,2002,6(20):422-423
[80]J Sawai,R Doi,Y Maekawa,et al.Indirect conductimetric assay of Antibacterial activities[J].Journal of Industrial Microbiology And Biotechnology,2002,29(5):296-298.
[81]Osamu Yamamoto,Miyako Komatsu,Jun Sawai,et al.Effect of lattice constant of zinc oxide on antibacterial characteristics[J].Journal of Materials Science:Materials in Medicine,2004,15(8):847-851
[82]谢鹏,张敏红.黄酮类化合物抑菌作用的研究进展[J].中国动物保健.2004.12:35-36
[83]井乐刚,张永忠,田璐.大豆异黄酮抑茵活性的研究[J].生物技术通讯.2004,15(5):476-479
[84]邵留,沈盎绿,郑曙明.斑地锦总黄酮的提取及抑菌作用[J],西南农业大学学报(自然科学版),2005,126(6):903-905
[85]Steven C Gllman and Thomas J.Rogers.Immunopharmacology[M].Fir-St ed.New Jersey:The Telfond press,1989,50-100
[86]Bond J S,Butler P E.Intracellular proteases.Annu Rev Biochem.1987,56:333-64
[87]Berfram G Katzung,ed.Basic and clinical pharmacology[M].Sixthed.East Norwalk:Appleton & lange,1995,895-878
[88]国家中医药管理局.中华本草第四册[M].上海出版社.1999,3576.
[89]Yanhua Lu,Chongwei Zhang,Peter Bucheli,et al.Citrus Flavonoids In Fruitand Traditional Chinese Medicinal Food Ingredients in China[J].Plant Foods for Human Nutrition,2006,61(2):55-63
[90]张雪梅,鞠玉琳,熊焕章.中药制剂的免疫药理学研究[J].中国兽医杂志,2008,6(44):44-45
[91]朱正美,刘辉.简明免疫学技术[M].北京:科学出版社,2002,205-206、220-221.
[92]王伟,白俊杰,劳海华,等.草鱼Mx蛋白基因的克隆与原核表达[J].中国水产科学,2003,10(5):366-368
[93]马建国,于江.槐白皮对机体免疫功能影响的研究[J].山东中医杂志,2002,21(9):557-559
[94]F Cappello.Is CDIa involved in antitumour immune responses during carcinogenesis[J].British Journal of Cancer,2004,90(23):938
[95]张兰玲,孟济明,陈海生,等.点腺过路黄正丁醇提取部分对小鼠免疫反应的影响[J].中药材,2002,25(12):888-890
[96]马晓彤,王庭欣,蒋东升,等.一种人参保健品功效作用的评价[J].卫生毒理学杂志,2003,17(1):61-62
[97]阎莉,郑作文,卫智权.广西藤茶总黄酮对正常小鼠免疫功能的影响[J].中国药物应用与监测,2008,5(4):5-7
[98]李小定,吴谋成,曾晓波,等.灰树花多糖粗品与纯品的抗肿瘤作用及对免疫功能的影响[J].营养学报,2003,25(1):7-9
[99]徐叔云,卞如镰,陈修.药理实验方法学(第三版)[M].北京:人民卫生出版社,2002.
[100]朱立平,陈学清.免疫学常用实验方法[M].北京:人民军医出版社,2000:175-180.
[101]戴伟娟,司端运,苏艾兰,等.无花果多糖对小鼠迟发型超敏反应的影响[J].济宁医学院学报,1999,22(4):26-27
[102]伍章保,沈玉先,魏伟,等.富硒酵母对免疫功能低下小鼠DTH的影响[J].中国药理学通报,1999,12(2):182-184
[103]胡相光,洪净,尹华熙,等.凉血清营颗粒对炎症及迟发型超敏反应的影响[J].中药药理与临床,2004,20(1):22-24
[104]杨保华,陈德宇,肖顺汉.消风胶囊对小鼠迟发型超敏反应的影响及其作用机制研究[J].中国中西医结合皮肤性病学杂志,2003,2(1):13-15
[105]张立煌,姚航平,曹雪涛,等.吡喹酮联合IL-18基因治疗血吸虫感染小鼠对免疫功能的影响[J].中国微生物与免疫学杂志,2001,21(1):87-90
[106]张丽丽,张竞逵,聂宏刚,等.不同浓度细辛提取物体外免疫抑制作用的实验研究[J].中华中西医杂志,2003,4(4):481-482
[107]张晓君,祝晨,胡黎,等.党参多糖对小鼠免疫和造血功能的影响[J].中药新药与临床药理,2003,14(14):174-176
[108]李桂峰,康裕财,孙际佳,等.酵母多糖对赤眼蹲非特异性免疫机能的影响[J].中山大学学报(自然科学版),2003,7(4):55-58
[109]林慧彬,林建强,杭建群,等.山东产四种冤丝子免疫增强作用的比较研究[J].中西医结合学报,2003,1(1):51-53
[110]石学魁,张晓莉,宋宝辉,等.五味消毒饮增强小鼠免疫功能的研究[J].细胞与分子免疫学杂志,2000,16(1):85
[111]沈关心,周汝麟.现代免疫学实验技术[M].武汉:湖北科学技术出版社,1998,255-256.
[112]Claudio Fiocchi,Kenneth R.Youngman,Belinda Yen-Lieberman,etal.Modulation of intestinal immune reactivity by interleukin 2Phenotypic and functional analysis of lymphokine activated kilLer cells from human intestinal mucosa[J].Digestive Diseases and Sciences(Historical Archive),1988,33(10):1305-1315
[113]Ar Neng Lai,Joseph C.K.Leung,Fernand Mac-Moune Lai,et al.Tlymphocyte activation in IgA nephropathy:Serum-soluble interleukin 2 receptor level,interleukin 2production,and interleukin 2 receptor expression by cultured lymphocytes[J].Journal of Clinic Al Immunology(Historical Archive),1989,9(6):485-492
[114]Lucie M.Heinzerling Karsten Feige,Stefan Rieder,Margarete K.Akens,et al.Tumor regression induced by intratumoral injection of DNA coding for human interleukin 12into melanoma metastases in gray horses[J].Journal of Molecular Medicine,2001,78(12):692-702
[115]A.Fayyazi,S.Schweyer,A.Soruri,et al.R.Parwaresch.Tlymphocytes and altered keratinocytes express interferon-γ and interleukin 6 in lichen planus[J].Archives of Dermatological Research,1999,291(9):485-490
[116]M.Yamada,Y.Arao,A.Hatano.S.Ni.Effect of recombinant mouse interferon-β on acute and latent herpes simplex infection in mice[J].Archives of Virology(Historical Archive),1988,99(1-2):101-109
[117]Y.Uchida,K.Ohba,A.Ogawa.T.Muraki.Protein kinase C mediates rumor necrosis factor-αinduced inhibition of obese gene expression and leptin secretion in brown adipocytes[J].Naunyn-Schmiedeberg's Archives of Pharmacology,1999,360(6):691-698
[118]刘清池,梁冰.中药诱生细胞因子研究进展[J].中医杂志,1997,38(12):47
[119]李宏梅、李祥瑞、郭慧君.白细胞介素-2的研究进展及应用[J].山东畜牧兽医,2002(5):38-40
[120]Yasuhiro Kodera,Hayao Nakanishi,Seiji Ito,et al.Quantitative detection of disseminated cancer cells in the greater omentum of gastric carcinoma patients with real-time RT-PCR:a comparison with peritoneal lavage cytology[J].Gastric Cancer,2002,5(2):69-70