大豆肽的制备及其免疫调节作用的研究
摘要
论文主要研究大豆肽的制备及其免疫调节功能,并初步探讨大豆肽免疫活性与其电荷及相对分子质量之间的关系。
通过单因素实验制备了不同水解条件下水解的大豆肽,采用SP—Sephadex C-25阳离子交换树脂测定水解肽正电荷肽含量,高效液相测定其相对分子质量的分布,发现水解肽的相对分子质量主要集中在1000以下,说明小肽是水解物中的主要成分。通过淋巴细胞增殖和巨噬细胞吞噬中性红实验,测得大豆肽对小鼠淋巴细胞增殖和巨噬细胞吞噬能力均有一定的促进作用,且大豆肽免疫活性大小与正电荷肽含量的相关性较好,而与平均分子量的相关性不大。其中,Alcalase为水解酶,酶用量为o.02g/g蛋白,底物浓度为5%,温度60℃,pH值8.0,水解时间225min,活性最高。
通过给昆明种雄性小鼠(5.6周龄)连续3 d注射40mg/Kg的环磷酰胺,并灌胃16mg/Kg、40mg/Kg、80mg/Kg的大豆肽,发现大豆肽不仅能显著提高正常小鼠免疫功能,而且能显著提高环磷酰胺致免疫低下小鼠的脏器指数,提高脾脏淋巴细胞转化率、抗体生成细胞含量及血清溶血素含量(P<0.05)。
用60mg/Kg的D-半乳糖连续42 d腹腔注射昆明种雄性小鼠(5.6周龄),同时灌胃16mg/Kg、40mg/Kg的大豆肽,结果表明,不同剂量大豆肽均能提高小鼠脏器指数,脾脏淋巴细胞转化率、抗体生成细胞含量及血清溶血素含量。
The aim of this study was to investigate the preparation and immunological regulation of soy peptides and the pertinency of the content of positively charged peptides, molecular weight and immunocompetence by preparing soy peptides hydrolyzed by different conditions.
Soy peptides hydrolyzed by different conditions. The content of positively charged peptides was determined by SP-Sephadex C-25 ionic-exchange resins.
Molecular weight of the hydrolydates were distributed under 1000. Small peptides were the main component. Soy peptides could stimulate lymphocyte proliferative and enhance the phagocytic activity of macrophages. The pertinency of immunocompetence and the content of positively charged peptides were satisfied, but molecular weight had only weak influence on immunocompetence. The optimum immunocompetence were determined: dosages of enzyme 0.02g/g protein, 60°C, pH8.0, substratum concentration 5%, hydrolysis time 225min.
KM mice (mole,5-6 weeks) were injected by 40mg/Kg BW cyclophosphamide through abdomen for 3 days, and having 16,40,80mg/Kg soy peptides. The result indicated that soy peptides could strengthen the normal mice immune function and enhanced the spleen index, thymus index, SI, PFC and HC50 of immuned hypofunctional mice (P<0.05).
KM mice (mole,5-6 weeks) were injected by 60 mg/Kg BW D-galactose through abdomen for 42 days and having 16,40 mg/Kg soy peptides. The result indicated that soy peptides could strengthen the mice immune function and enhance the spleen index, thymus index, SI, PFC and HC5o of mice.
通过单因素实验制备了不同水解条件下水解的大豆肽,采用SP—Sephadex C-25阳离子交换树脂测定水解肽正电荷肽含量,高效液相测定其相对分子质量的分布,发现水解肽的相对分子质量主要集中在1000以下,说明小肽是水解物中的主要成分。通过淋巴细胞增殖和巨噬细胞吞噬中性红实验,测得大豆肽对小鼠淋巴细胞增殖和巨噬细胞吞噬能力均有一定的促进作用,且大豆肽免疫活性大小与正电荷肽含量的相关性较好,而与平均分子量的相关性不大。其中,Alcalase为水解酶,酶用量为o.02g/g蛋白,底物浓度为5%,温度60℃,pH值8.0,水解时间225min,活性最高。
通过给昆明种雄性小鼠(5.6周龄)连续3 d注射40mg/Kg的环磷酰胺,并灌胃16mg/Kg、40mg/Kg、80mg/Kg的大豆肽,发现大豆肽不仅能显著提高正常小鼠免疫功能,而且能显著提高环磷酰胺致免疫低下小鼠的脏器指数,提高脾脏淋巴细胞转化率、抗体生成细胞含量及血清溶血素含量(P<0.05)。
用60mg/Kg的D-半乳糖连续42 d腹腔注射昆明种雄性小鼠(5.6周龄),同时灌胃16mg/Kg、40mg/Kg的大豆肽,结果表明,不同剂量大豆肽均能提高小鼠脏器指数,脾脏淋巴细胞转化率、抗体生成细胞含量及血清溶血素含量。
The aim of this study was to investigate the preparation and immunological regulation of soy peptides and the pertinency of the content of positively charged peptides, molecular weight and immunocompetence by preparing soy peptides hydrolyzed by different conditions.
Soy peptides hydrolyzed by different conditions. The content of positively charged peptides was determined by SP-Sephadex C-25 ionic-exchange resins.
Molecular weight of the hydrolydates were distributed under 1000. Small peptides were the main component. Soy peptides could stimulate lymphocyte proliferative and enhance the phagocytic activity of macrophages. The pertinency of immunocompetence and the content of positively charged peptides were satisfied, but molecular weight had only weak influence on immunocompetence. The optimum immunocompetence were determined: dosages of enzyme 0.02g/g protein, 60°C, pH8.0, substratum concentration 5%, hydrolysis time 225min.
KM mice (mole,5-6 weeks) were injected by 40mg/Kg BW cyclophosphamide through abdomen for 3 days, and having 16,40,80mg/Kg soy peptides. The result indicated that soy peptides could strengthen the normal mice immune function and enhanced the spleen index, thymus index, SI, PFC and HC50 of immuned hypofunctional mice (P<0.05).
KM mice (mole,5-6 weeks) were injected by 60 mg/Kg BW D-galactose through abdomen for 42 days and having 16,40 mg/Kg soy peptides. The result indicated that soy peptides could strengthen the mice immune function and enhance the spleen index, thymus index, SI, PFC and HC5o of mice.
引文
[1]陈慰峰主编.医学免疫学[M].北京:人民卫生出版社,2002:1.
[2]周光炎.免疫学原理[M].上海:上海科学技术文献出版社,2001:12-14.
[3] Emanuele Smacchil, Marco Gobbetti. Bioactice peptides in dairy products: synthesis and interaction with proteolytic enzymes[J]. Food Microbiology,2000,17:129-141.
[4] Hannu Korhonen, Anne Pihlanto-Leppala, Pirjo Rantamaki and Tuomo Tupasela. Impact of processing on bioactice proteins and peptides[J].Food Science and Technology,1998,9:307-319.
[5]曹明耀,刘延菊.几种免疫活性肽的研究进展[J]中国基层医药,2004,11(2):238-239.
[6]曹文红,章超桦,秦小明.食品蛋白来源生物活性肽[J].粮食与油脂,2007,2:2-6.
[7]王秋韫,庞广昌,陈庆森.免疫活性肽的研究进展与展望[J].食品科学,2002,23(7):136-139.
[8]曾晓波,王海英,林永成等.食物中的生物活性肽,生物活性及研究进展[J].食品工业科技,2004,25(4):151-155.
[9] Hadden J W.Immunotheraphy of human immunodeficiency virus infection[J]. Trends Pharmaceutical Sci.,1991,12:107-111.
[10] M.Yoshikawa, M.Takahashi, Immunomodulating peptide derived from soybean protein Annals[M]. New york Acad. Of Sci.,1993,375-376.
[11] M. Takahashi,M.Yoshikawa.Isolation and Characterization of oryzatensin: a novel bioactive peptide with ileum---contracting and immunmodulating activites derived from rice albumin [J]. Biochem. and Molecular Biol. International, 1994,33(6):1151-1158.
[12]杨小军,左伟勇,陈伟华等.灌喂大豆蛋白胃蛋白酶酶解物对大鼠免疫功能的影响[J].畜牧兽医学报,2005,36(4):348-351.
[13]蒲首丞,王金水.生物活性肽的制备与应用[J].粮食加工,2005, 5: 49-51.
[14]程云辉,文新华.生物活性肽制备的研究进展[J].食品与机械,2001,4:4-7.
[15]张琦.生物活性肽制备的研究[J].畜牧兽医杂志,2003,22(3):20-21.
[16]薛庆善主编.体外培养的原理与技术[M].北京:科学出版社,2001:18.
[17] Alder-Nissen. Determination of the degree of hydrolysis of food protein hydrolyzates by trinitrobenzene sulfonnic acid[J].Agric Food Chem,1989(27):17-20.
[18]曲永洵.大豆多肽的特性及其应用[J].中国油脂.1996,21(2):3-5.
[19]潘进权,刘耕.大豆多肽研究概况[J].中国调味品,2003,288(2):6-10.
[20]李玉珍,林亲录,肖怀秋等.大豆多肽特性及其应用研究现状[J].中国食品添加剂,2005,6:91-94.
[21]张延坤,刘炳智.大豆肤在食品工业中的应用[J].食品工业,1997,3:5-60.
[22]吴建平,丁霄霖.大豆降压肽的研制(B)-酶E作用条件的优化[J].中国油脂,1998,23,(3):6-8.
[23]吴建平,潘文彪.乳源生物活性肽的研究概述[J].中国乳品工业,1999,27(1):12-15.
[24]焦迎春,郑晓冬.活性肽在食品中的应用[J].粮油加工与食品机械,2002,8:35-37.
[25]陈历俊.大豆活性肽的研制与应用----大豆深加工研究系列之一东北农业大学博士学位论文.1998.
[26]左伟勇,孟婷,陈伟华.植物蛋白源活性肽研究进展[J].动物营养,2007,1:22-24.
[27]高长城,胡锐,李煌馨.大豆肽对增强体能的作用[J].大豆通报,2001,2:24.
[28]卢阳,王凤翼,孔繁东等.大豆蛋白酶水解物抗氧化性的研究[J].大连轻工业学院学报,2001,12(20)25-29.
[29] Hua-ming Chen,Koji Muramoto. Fumauchi.Structural Analysis of Antioxdative Peptide from Soybean-Conglycinin[J].Agric Food Chem,1995,43:574-578.
[30] Hua-ming Chen,Koji Muramoto,Fumio Yamauchi et. Antioxdative Peptide of Histidine-Containing peptides Disigned from Peptide Fragments Found in the Digests of a Soybean Protein[J]. Agric Food Chem, 1998,46:49-53.
[31]潘翠玲,陈伟华,邹思湘等.添喂大豆蛋白酶解物对早期断乳仔猪免疫组织及器官的影响[J].江苏农业学报,2005,21(3): 212-215.
[32] Wenyi Wang and Elvira Gonzalez de Mejia. A frontier in soy bioactive peptides that may prevent age-related chronic diseases[J]. Food Science and Food Satety.2005,4:63-78.
[33] Kraus,J.,and Franz,G. Immunomodulating effects of polysaccharides from medicinal plants. Inmicrobial Infections,eds.Friedmam,H.,K1ein,T.W.,and Yamaguchi ,H., Penum Press,New York,1992:299-308.
[34] Ooi V.E. C. and Liu F. Immunomodulation and anti-cancer activity of polysaccharide-protein complexes[J].Current Medicinal Chemistry.2000,7:715-29.
[35] Dong Y., Yang M.M. and Kwan C.Y. In vitro inhibition of proliferation of HL-60 cells by tetrandrine and peptide derived from Chinese medicinal herbs[J].Life Sci.1997,60:35-40.
[36] Bohn, J. A., Bemiller,J.N.(1-3)-β-D-Glucans as biological response modifiera:a review of structure-functional activity relationships[J].Carbohydrate polymers,1995,28:3-14.
[37] Tzianabos A. Polysaccharide immunomodulators as therapeutic agentsatructural aspects and biologic function[J].Clinical Microbiology Reviews,2000,13: 23-33.
[38]张亚飞,乐国伟,施用晖等.小麦蛋白Alcalase水解物免疫活性肽的研究[J].食品与机械,2006,(22),3:44-46.
[39]刘健敏,钟芳,麻建国.大豆生理活性肽的研究(I)[J].无锡轻工大学学报,2004,23(3):41-45.
[40] Hua-Ming Chen, Koji Muramoto, Funio Yamauchi. Structural analysis of antioxidative peptides from Soybeanβ-conglycinin[J].J Agric Food Chem,1995,43:574-578.
[41] Flavia M.Netto and Maria Antonia M.Galeazzi. Production and characterization of enzymatic hydrolysate from soy protein isolate[J]. Academic Press,1998,31:624-631.
[42] Adler-Nissen, J., Enzymic hydrolysis of food proteins[M].Elsevier applied science publishers, London and New York, 1986:170-180.
[43] Adamson,Nicholas J.and Reynolds, Eric C.Characterization of casein phosphopeptides prepared using alcalase:Determination of enzyme specificity[J].Enzyme and Microbial Technology,1996,19(3):202-207.
[44]孙旸,大豆肽的制备分离及特性研究.吉林农业大学硕士论文,2004:23.
[45] Emanuele Smacchil, Marco Gobbetti. Bioactive peptides in dairy products:synthesis and interaction with proteolytic enzymes[J].Food Microbiology,2000,17:129-141.
[46]陈志阳,王国华,杨敏等.一种简便、快速检测淋巴细胞增殖的方法[J].中国生化药物杂志,2000,21(5):245-247.
[47]刘民,马华,李柏青.MTT法检测小鼠淋巴细胞增殖性反应探讨[J].中国实验动物学杂志,1999,9(3):
[48]张传开,袁盛榕,刘进学等.冬虫夏草和中国拟青霉水提取物对小鼠免疫功能的影响[J].首都医科大学学报,1999,20(3):17-20.
[49]黎观红,乐国伟,施用晖等.利用自建的活性肽数据库搜寻食物蛋白质中潜在的生物活性肽[J].食品与发酵工业,2004,30(1):85-88.
[50]毛跟年,许牡丹主编.功能食品生理特性与检测技术[M].北京:化学工业出版社,2005:391-392.
[51]戴伟娟,司端运,仲伟法.无花果多糖对小鼠单核吞噬细胞吞噬功能影响的研究[J].中医药学刊,2002,20(1):98-99.
[52]汪家政,范明主编.蛋白质技术手册[M].北京,2004:190.
[53] Emanuele Smacchil, Marco Gobbetti. Bioactive peptides in dairy products:synthesis and interaction with proteolytic enzymes[J].Food Microbiology,2000,17:129-141.
[54]Lersch C.,Zeuner M.,Bauer A.,et al. Stimulation of the immune response in outpatients with hepatocellular carcinomas by low dose of cyclophosphamide(LDCY),Echinacea Purpurea Extract(Echinacin) and thymostihmulin[J]. Arch Geschwulstforsch, 1990;60:379-383.
[55] Dumont F.Desrtruction and regeneration of lymphocyte population in the mouse spleen after cyclophosphamide treatment[J]. International Archives in Allergy Immunology,1974:47:110-123.
[56]李祥华,张家均,王文英,等.胶艾汤对小鼠免疫功能的影响[J],时珍国医国药, 2005, 16(5): 379.
[57]郑国华,李会庆.大蒜提取液对小鼠脾脏及血清溶血素抗体影响[J],福建中医学院学报, 2006, 16(4): 33.
[58]樊俊杰,贾正平,谢景文等.瑞香狼毒多糖对环磷酰胺处理小鼠免疫功能的影响[J].西北国防医学杂志,2002,21(4):263-265.
[59]许华,李静梅.欧瑞活血素对环磷酰胺模型小鼠免疫功能的影响[J].中国新药杂志,2002,11(11):853-858.
[60]Wick G,Jansen2Durr P ,Berger P. Disease of aging[J ] . Vaccine ,2000 ,18(16) :1567-1583.
[61]Pawelec G,Adibzadeh M,Pohla H ,et al . Immunosenescence :ageing of the immune system[J ] . Immunol Today. 1995 ,16 (9) :420-422.
[62]陈美珍,余杰,郭慧敏.大豆分离蛋白酶解物清除羟自由基作用的研究[J].食品科学, 2002, 23(1): 43-47.
[63]黄莉,江连洲,朱秀清.大豆蛋白抗氧性肽的研究[J].大豆通报,2005,(5): 20-21.
[64]刘明,倪辉,吴永沛.大豆抗氧化活性肽的研究进展[J].食品科学,2006,27(12)897-900.
[65]穆运转.自由基清除剂的辐射防护作用[J].中华放射医学与防护杂志, 1990, 10(4): 280-282.
[66]李燕,刘晓丽,裴素萍等。银耳多糖对实验衰老模型小鼠免疫功能的影响[J].中国临床营养杂志,2005,8:228-231.
[2]周光炎.免疫学原理[M].上海:上海科学技术文献出版社,2001:12-14.
[3] Emanuele Smacchil, Marco Gobbetti. Bioactice peptides in dairy products: synthesis and interaction with proteolytic enzymes[J]. Food Microbiology,2000,17:129-141.
[4] Hannu Korhonen, Anne Pihlanto-Leppala, Pirjo Rantamaki and Tuomo Tupasela. Impact of processing on bioactice proteins and peptides[J].Food Science and Technology,1998,9:307-319.
[5]曹明耀,刘延菊.几种免疫活性肽的研究进展[J]中国基层医药,2004,11(2):238-239.
[6]曹文红,章超桦,秦小明.食品蛋白来源生物活性肽[J].粮食与油脂,2007,2:2-6.
[7]王秋韫,庞广昌,陈庆森.免疫活性肽的研究进展与展望[J].食品科学,2002,23(7):136-139.
[8]曾晓波,王海英,林永成等.食物中的生物活性肽,生物活性及研究进展[J].食品工业科技,2004,25(4):151-155.
[9] Hadden J W.Immunotheraphy of human immunodeficiency virus infection[J]. Trends Pharmaceutical Sci.,1991,12:107-111.
[10] M.Yoshikawa, M.Takahashi, Immunomodulating peptide derived from soybean protein Annals[M]. New york Acad. Of Sci.,1993,375-376.
[11] M. Takahashi,M.Yoshikawa.Isolation and Characterization of oryzatensin: a novel bioactive peptide with ileum---contracting and immunmodulating activites derived from rice albumin [J]. Biochem. and Molecular Biol. International, 1994,33(6):1151-1158.
[12]杨小军,左伟勇,陈伟华等.灌喂大豆蛋白胃蛋白酶酶解物对大鼠免疫功能的影响[J].畜牧兽医学报,2005,36(4):348-351.
[13]蒲首丞,王金水.生物活性肽的制备与应用[J].粮食加工,2005, 5: 49-51.
[14]程云辉,文新华.生物活性肽制备的研究进展[J].食品与机械,2001,4:4-7.
[15]张琦.生物活性肽制备的研究[J].畜牧兽医杂志,2003,22(3):20-21.
[16]薛庆善主编.体外培养的原理与技术[M].北京:科学出版社,2001:18.
[17] Alder-Nissen. Determination of the degree of hydrolysis of food protein hydrolyzates by trinitrobenzene sulfonnic acid[J].Agric Food Chem,1989(27):17-20.
[18]曲永洵.大豆多肽的特性及其应用[J].中国油脂.1996,21(2):3-5.
[19]潘进权,刘耕.大豆多肽研究概况[J].中国调味品,2003,288(2):6-10.
[20]李玉珍,林亲录,肖怀秋等.大豆多肽特性及其应用研究现状[J].中国食品添加剂,2005,6:91-94.
[21]张延坤,刘炳智.大豆肤在食品工业中的应用[J].食品工业,1997,3:5-60.
[22]吴建平,丁霄霖.大豆降压肽的研制(B)-酶E作用条件的优化[J].中国油脂,1998,23,(3):6-8.
[23]吴建平,潘文彪.乳源生物活性肽的研究概述[J].中国乳品工业,1999,27(1):12-15.
[24]焦迎春,郑晓冬.活性肽在食品中的应用[J].粮油加工与食品机械,2002,8:35-37.
[25]陈历俊.大豆活性肽的研制与应用----大豆深加工研究系列之一东北农业大学博士学位论文.1998.
[26]左伟勇,孟婷,陈伟华.植物蛋白源活性肽研究进展[J].动物营养,2007,1:22-24.
[27]高长城,胡锐,李煌馨.大豆肽对增强体能的作用[J].大豆通报,2001,2:24.
[28]卢阳,王凤翼,孔繁东等.大豆蛋白酶水解物抗氧化性的研究[J].大连轻工业学院学报,2001,12(20)25-29.
[29] Hua-ming Chen,Koji Muramoto. Fumauchi.Structural Analysis of Antioxdative Peptide from Soybean-Conglycinin[J].Agric Food Chem,1995,43:574-578.
[30] Hua-ming Chen,Koji Muramoto,Fumio Yamauchi et. Antioxdative Peptide of Histidine-Containing peptides Disigned from Peptide Fragments Found in the Digests of a Soybean Protein[J]. Agric Food Chem, 1998,46:49-53.
[31]潘翠玲,陈伟华,邹思湘等.添喂大豆蛋白酶解物对早期断乳仔猪免疫组织及器官的影响[J].江苏农业学报,2005,21(3): 212-215.
[32] Wenyi Wang and Elvira Gonzalez de Mejia. A frontier in soy bioactive peptides that may prevent age-related chronic diseases[J]. Food Science and Food Satety.2005,4:63-78.
[33] Kraus,J.,and Franz,G. Immunomodulating effects of polysaccharides from medicinal plants. Inmicrobial Infections,eds.Friedmam,H.,K1ein,T.W.,and Yamaguchi ,H., Penum Press,New York,1992:299-308.
[34] Ooi V.E. C. and Liu F. Immunomodulation and anti-cancer activity of polysaccharide-protein complexes[J].Current Medicinal Chemistry.2000,7:715-29.
[35] Dong Y., Yang M.M. and Kwan C.Y. In vitro inhibition of proliferation of HL-60 cells by tetrandrine and peptide derived from Chinese medicinal herbs[J].Life Sci.1997,60:35-40.
[36] Bohn, J. A., Bemiller,J.N.(1-3)-β-D-Glucans as biological response modifiera:a review of structure-functional activity relationships[J].Carbohydrate polymers,1995,28:3-14.
[37] Tzianabos A. Polysaccharide immunomodulators as therapeutic agentsatructural aspects and biologic function[J].Clinical Microbiology Reviews,2000,13: 23-33.
[38]张亚飞,乐国伟,施用晖等.小麦蛋白Alcalase水解物免疫活性肽的研究[J].食品与机械,2006,(22),3:44-46.
[39]刘健敏,钟芳,麻建国.大豆生理活性肽的研究(I)[J].无锡轻工大学学报,2004,23(3):41-45.
[40] Hua-Ming Chen, Koji Muramoto, Funio Yamauchi. Structural analysis of antioxidative peptides from Soybeanβ-conglycinin[J].J Agric Food Chem,1995,43:574-578.
[41] Flavia M.Netto and Maria Antonia M.Galeazzi. Production and characterization of enzymatic hydrolysate from soy protein isolate[J]. Academic Press,1998,31:624-631.
[42] Adler-Nissen, J., Enzymic hydrolysis of food proteins[M].Elsevier applied science publishers, London and New York, 1986:170-180.
[43] Adamson,Nicholas J.and Reynolds, Eric C.Characterization of casein phosphopeptides prepared using alcalase:Determination of enzyme specificity[J].Enzyme and Microbial Technology,1996,19(3):202-207.
[44]孙旸,大豆肽的制备分离及特性研究.吉林农业大学硕士论文,2004:23.
[45] Emanuele Smacchil, Marco Gobbetti. Bioactive peptides in dairy products:synthesis and interaction with proteolytic enzymes[J].Food Microbiology,2000,17:129-141.
[46]陈志阳,王国华,杨敏等.一种简便、快速检测淋巴细胞增殖的方法[J].中国生化药物杂志,2000,21(5):245-247.
[47]刘民,马华,李柏青.MTT法检测小鼠淋巴细胞增殖性反应探讨[J].中国实验动物学杂志,1999,9(3):
[48]张传开,袁盛榕,刘进学等.冬虫夏草和中国拟青霉水提取物对小鼠免疫功能的影响[J].首都医科大学学报,1999,20(3):17-20.
[49]黎观红,乐国伟,施用晖等.利用自建的活性肽数据库搜寻食物蛋白质中潜在的生物活性肽[J].食品与发酵工业,2004,30(1):85-88.
[50]毛跟年,许牡丹主编.功能食品生理特性与检测技术[M].北京:化学工业出版社,2005:391-392.
[51]戴伟娟,司端运,仲伟法.无花果多糖对小鼠单核吞噬细胞吞噬功能影响的研究[J].中医药学刊,2002,20(1):98-99.
[52]汪家政,范明主编.蛋白质技术手册[M].北京,2004:190.
[53] Emanuele Smacchil, Marco Gobbetti. Bioactive peptides in dairy products:synthesis and interaction with proteolytic enzymes[J].Food Microbiology,2000,17:129-141.
[54]Lersch C.,Zeuner M.,Bauer A.,et al. Stimulation of the immune response in outpatients with hepatocellular carcinomas by low dose of cyclophosphamide(LDCY),Echinacea Purpurea Extract(Echinacin) and thymostihmulin[J]. Arch Geschwulstforsch, 1990;60:379-383.
[55] Dumont F.Desrtruction and regeneration of lymphocyte population in the mouse spleen after cyclophosphamide treatment[J]. International Archives in Allergy Immunology,1974:47:110-123.
[56]李祥华,张家均,王文英,等.胶艾汤对小鼠免疫功能的影响[J],时珍国医国药, 2005, 16(5): 379.
[57]郑国华,李会庆.大蒜提取液对小鼠脾脏及血清溶血素抗体影响[J],福建中医学院学报, 2006, 16(4): 33.
[58]樊俊杰,贾正平,谢景文等.瑞香狼毒多糖对环磷酰胺处理小鼠免疫功能的影响[J].西北国防医学杂志,2002,21(4):263-265.
[59]许华,李静梅.欧瑞活血素对环磷酰胺模型小鼠免疫功能的影响[J].中国新药杂志,2002,11(11):853-858.
[60]Wick G,Jansen2Durr P ,Berger P. Disease of aging[J ] . Vaccine ,2000 ,18(16) :1567-1583.
[61]Pawelec G,Adibzadeh M,Pohla H ,et al . Immunosenescence :ageing of the immune system[J ] . Immunol Today. 1995 ,16 (9) :420-422.
[62]陈美珍,余杰,郭慧敏.大豆分离蛋白酶解物清除羟自由基作用的研究[J].食品科学, 2002, 23(1): 43-47.
[63]黄莉,江连洲,朱秀清.大豆蛋白抗氧性肽的研究[J].大豆通报,2005,(5): 20-21.
[64]刘明,倪辉,吴永沛.大豆抗氧化活性肽的研究进展[J].食品科学,2006,27(12)897-900.
[65]穆运转.自由基清除剂的辐射防护作用[J].中华放射医学与防护杂志, 1990, 10(4): 280-282.
[66]李燕,刘晓丽,裴素萍等。银耳多糖对实验衰老模型小鼠免疫功能的影响[J].中国临床营养杂志,2005,8:228-231.