大豆凝集素受体标记及大豆凝集素对大鼠小肠上皮细胞的影响
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摘要
本研究分三个试验：1.以FITC-大豆凝集素为探针对SD大鼠肠上皮细胞系IEC-6表面的大豆凝集素受体进行标记,2.对本实验室纯化的大豆凝集素进行鉴定确定大豆凝集素的有效性,同时研究了培养基对大豆凝集素活性的影响。3.探讨了大豆凝集素对体外培养的大鼠小肠上皮细胞系(IEC-6)生长和迁移的影响。
IEC-6的同步化处理及其大豆凝集素受体的荧光标记。细胞的同步化处理：试验通过单克隆方法对细胞进行同步化处理,进行形态学观察并用MTT比色法测定其增殖能力。荧光标记实验：用10μg·ml-1的FITC-大豆凝集素冷丙酮固定的IEC-6细胞爬片孵育30min。
测定本实验室纯化大豆凝集素分子量、纯度、抗原特异性和凝集活性,并检测培养基对大豆凝集素凝血活性的影响。试验采用凝胶电泳方法对其分子量、纯度进行测定,用Western-blot方法对其鉴定,用红细胞凝集试验对其凝血活性进行检测,并将其与Sigma大豆凝集素标准品各指标进行比较。通过红细胞凝集实验检测并比较了大豆凝集素浓度均为1mg·ml-1的生长培养基和阳性对照的凝血活性。
大豆凝集素对细胞增殖和细胞迁移的影响。细胞增殖试验：分别以大豆凝集素浓度为0,0.5,1.0,2.0,3.0,4.0,5.0,6.0mg·ml-1的培养基对IEC-6进行处理,其中0 mg·ml-1组为对照组,并在0,12,24,48,60,72,84h用MTT比色试验测定细胞活性；细胞迁移试验：分别用大豆凝集素浓度为0,0.5,1.0,1.5mg·ml-1的培养基对铺满细胞的盖玻片进行处理其中0mg·ml-1组为对照组,并在0d、1d、2d、3d观察上皮细胞的迁移数量。
同步化处理结果：经过同步化处理细胞所有细胞生长形态单一,有1-2个核仁,细胞排列整齐,呈多边形铺路石状；生长曲线测定结果显示,细胞生命力旺盛,接种后12h后可全部贴壁生长,24h后进入对数期,48h进入平台期直到84h进入衰亡期。荧光标记结果为强阳性,说明细胞含有大量的大豆凝集素受体。
大豆凝集素鉴定结果：试验测得本实验室纯化大豆凝集素分子量为30kDa,纯度在99%以上,Western-blot结果为阳性,凝集效价为1/210,各指标与Sigma标准品之间均未见差异且与理论值相同。红细胞凝集实验显示大豆凝集素浓度为1mg·ml-1的生长培养基和阳性对照的血凝效价均为1/210。说明试验所用大豆凝集素与标准品没有显著差异,所用培养基对大豆凝集素的活性没有显著影响。
细胞增殖试验结果：各剂量组细胞活性随时间呈现先升高后下降的趋势,在72h出现最高值,84h后开始下降。同一测量时间,随着凝集素含量的升高,细胞活性呈现先升高后降低的趋势,并在0.5,1.0mg·ml-1显著高于对照组(P<0.05),2.0mg·ml-1虽然高于对照组但差异不显著(P>0.05),3.0mg·ml-1组显著低于对照组(P<0.05),4.0,5.0,6.0mg·ml-1组低于对照组且差异极显著(p<0.01)。说明低剂量大豆凝集素对细胞的增殖有一定促进作用,而当凝集素浓度高于一定范围后对细胞增殖有促进作用减弱,甚至对细胞增殖有抑制作用。细胞迁移试验结果,各剂量组与对照组细胞从盖玻片向周围迁移的数量随时间延长而增加；同一测量时间,随着大豆凝集素浓度增,细胞迁出数量呈现升高趋势,且各时间点当浓度为1.5 mg·ml-1时显著高于对照组和其他剂量组(P<0.05)。说明在试验浓度范围内,大豆凝集素可能促进细胞迁移。说明低浓度大豆凝集素可能促进细胞增殖,高浓度可能抑制细胞增殖,试验大豆凝集素浓度范围内对细胞迁移有促进作用。
综上所述,IEC-6细胞存在大量的大豆凝集素受体低浓度(<2mg·ml-1)的大豆凝集素能够促进IEC-6的增殖,但较高浓度(>2mg·ml-1)大豆凝集素对其增殖的促进作用减弱或抑制细胞增殖。在一定范围内(<1.5mg·ml-1),大豆凝集素对细胞的迁移有促进作用,且随浓度的增加而加强。
Three experiments were conducted. Firstly to label the soybean agglutinin receptors on the cells of intestinal epithelial cells line of SD-rat IEC-6, secondly to identify the purified soybean agglutinin, and evaluate the influence of the growth medium of the cell lines on the agglutinative activity of soybean agglutinin, and thirdly to investigate the effects of soybean agglutinin on the proliferation and immigration of IEC-6.
IEC-6 cells were synchronized using a monoclonal method with the morphology and the growth curve recorded. The labeling of soybean agglutinin was done after fixing the cells with cold acetone and incubating them with FITC-SBA for about half an hour at 37℃.
The molecular weight, purity and agglutinative activity of the soybean agglutinin purified in our laboratory were detected as well as the effect of growth medium on the agglutinative activity. SDS-PAGE gel electrophoresis was used to detect the molecular weight and purity of the soybean agglutinin purified, Western-blot for the antigenic specificity, and agglutination test for the agglutinative activity. The findings were then compared with the results of the Sigma standard Soybean agglutinin. The effect of growth medium on the agglutinative activity was also evaluated by comparing values of agglutination between the cell growth medium containing 1mg·ml-1 soybean agglutinin and the control group.
The effects of soybean agglutinin on the cell proliferation and cell immigration were assessed after treating IEC-6 cells in logarithmic phase with growth medium containing SBA with the concentration of 0,0.5,1.0,2.0,3.0,4.0,5.0,6.0 mg·ml-1, and set 0 mg·ml-1 group as control. Proliferation curve of the cells was detected by the means of MTT method. For the cell immigration test, cells were treated with the medium with different concentrations of soybean agglutinin, i.e.0,0.5,1.0,1.5 mg·ml-1 and the number of immigration cells in different groups recorded.
After synchronizing the growth of the cells, the growth curve showed that cells grew very well. At about 12h after seeding, the cells got into a logarithmic phase, from 48h to 84h the cells were in a plateau phase and after that they got into a decline phase. The results of labeling showed that there were a lot of soybean agglutinin receptors on the cells of IEC-6.
The molecular weight of soya bean agglutinin was 30.78kDa, the level of purity was 98% and the value of agglutination was 1/210. All of these were not significantly different with those of Sigma standard soybean agglutinin (P>0.05). The values of agglutination of growth medium group and the control group were both 1/210. It indicated that the components of the cell growth medium had no influence on the agglutinative activity.
With time, results of the proliferation test revealed that the number of activity in all cell groups rose at first and then declined later. After adding the growth medium at 72h, cell activity approached the summit, but begun to decline thereafter. In 0.5 and 1.0 mg·ml-1 groups the activity of cells was significantly higher than the control group (P<0.05), while significantly lower in the 3.0 mg·ml-1 (P<0.05), and the 4.0,5.0, 6.0mg·ml-1 groups (P<0.01), respectively. This indicated that low concentration of soybean agglutinin could enhance the proliferation of the cells and that high concentration of SBA does not or even inhibit it. Cells immigrated from the cover-glasses in all groups with the number of immigration becoming higher with the increase of the soybean agglutinin concentration. At the highest concentration group, the numbers of cells numbers were significantly higher than all the other groups (P<0.05). Using this concentration of soybean agglutinin could enhance the immigration of the cell.
IEC-6的同步化处理及其大豆凝集素受体的荧光标记。细胞的同步化处理：试验通过单克隆方法对细胞进行同步化处理,进行形态学观察并用MTT比色法测定其增殖能力。荧光标记实验：用10μg·ml-1的FITC-大豆凝集素冷丙酮固定的IEC-6细胞爬片孵育30min。
测定本实验室纯化大豆凝集素分子量、纯度、抗原特异性和凝集活性,并检测培养基对大豆凝集素凝血活性的影响。试验采用凝胶电泳方法对其分子量、纯度进行测定,用Western-blot方法对其鉴定,用红细胞凝集试验对其凝血活性进行检测,并将其与Sigma大豆凝集素标准品各指标进行比较。通过红细胞凝集实验检测并比较了大豆凝集素浓度均为1mg·ml-1的生长培养基和阳性对照的凝血活性。
大豆凝集素对细胞增殖和细胞迁移的影响。细胞增殖试验：分别以大豆凝集素浓度为0,0.5,1.0,2.0,3.0,4.0,5.0,6.0mg·ml-1的培养基对IEC-6进行处理,其中0 mg·ml-1组为对照组,并在0,12,24,48,60,72,84h用MTT比色试验测定细胞活性；细胞迁移试验：分别用大豆凝集素浓度为0,0.5,1.0,1.5mg·ml-1的培养基对铺满细胞的盖玻片进行处理其中0mg·ml-1组为对照组,并在0d、1d、2d、3d观察上皮细胞的迁移数量。
同步化处理结果：经过同步化处理细胞所有细胞生长形态单一,有1-2个核仁,细胞排列整齐,呈多边形铺路石状；生长曲线测定结果显示,细胞生命力旺盛,接种后12h后可全部贴壁生长,24h后进入对数期,48h进入平台期直到84h进入衰亡期。荧光标记结果为强阳性,说明细胞含有大量的大豆凝集素受体。
大豆凝集素鉴定结果：试验测得本实验室纯化大豆凝集素分子量为30kDa,纯度在99%以上,Western-blot结果为阳性,凝集效价为1/210,各指标与Sigma标准品之间均未见差异且与理论值相同。红细胞凝集实验显示大豆凝集素浓度为1mg·ml-1的生长培养基和阳性对照的血凝效价均为1/210。说明试验所用大豆凝集素与标准品没有显著差异,所用培养基对大豆凝集素的活性没有显著影响。
细胞增殖试验结果：各剂量组细胞活性随时间呈现先升高后下降的趋势,在72h出现最高值,84h后开始下降。同一测量时间,随着凝集素含量的升高,细胞活性呈现先升高后降低的趋势,并在0.5,1.0mg·ml-1显著高于对照组(P<0.05),2.0mg·ml-1虽然高于对照组但差异不显著(P>0.05),3.0mg·ml-1组显著低于对照组(P<0.05),4.0,5.0,6.0mg·ml-1组低于对照组且差异极显著(p<0.01)。说明低剂量大豆凝集素对细胞的增殖有一定促进作用,而当凝集素浓度高于一定范围后对细胞增殖有促进作用减弱,甚至对细胞增殖有抑制作用。细胞迁移试验结果,各剂量组与对照组细胞从盖玻片向周围迁移的数量随时间延长而增加；同一测量时间,随着大豆凝集素浓度增,细胞迁出数量呈现升高趋势,且各时间点当浓度为1.5 mg·ml-1时显著高于对照组和其他剂量组(P<0.05)。说明在试验浓度范围内,大豆凝集素可能促进细胞迁移。说明低浓度大豆凝集素可能促进细胞增殖,高浓度可能抑制细胞增殖,试验大豆凝集素浓度范围内对细胞迁移有促进作用。
综上所述,IEC-6细胞存在大量的大豆凝集素受体低浓度(<2mg·ml-1)的大豆凝集素能够促进IEC-6的增殖,但较高浓度(>2mg·ml-1)大豆凝集素对其增殖的促进作用减弱或抑制细胞增殖。在一定范围内(<1.5mg·ml-1),大豆凝集素对细胞的迁移有促进作用,且随浓度的增加而加强。
Three experiments were conducted. Firstly to label the soybean agglutinin receptors on the cells of intestinal epithelial cells line of SD-rat IEC-6, secondly to identify the purified soybean agglutinin, and evaluate the influence of the growth medium of the cell lines on the agglutinative activity of soybean agglutinin, and thirdly to investigate the effects of soybean agglutinin on the proliferation and immigration of IEC-6.
IEC-6 cells were synchronized using a monoclonal method with the morphology and the growth curve recorded. The labeling of soybean agglutinin was done after fixing the cells with cold acetone and incubating them with FITC-SBA for about half an hour at 37℃.
The molecular weight, purity and agglutinative activity of the soybean agglutinin purified in our laboratory were detected as well as the effect of growth medium on the agglutinative activity. SDS-PAGE gel electrophoresis was used to detect the molecular weight and purity of the soybean agglutinin purified, Western-blot for the antigenic specificity, and agglutination test for the agglutinative activity. The findings were then compared with the results of the Sigma standard Soybean agglutinin. The effect of growth medium on the agglutinative activity was also evaluated by comparing values of agglutination between the cell growth medium containing 1mg·ml-1 soybean agglutinin and the control group.
The effects of soybean agglutinin on the cell proliferation and cell immigration were assessed after treating IEC-6 cells in logarithmic phase with growth medium containing SBA with the concentration of 0,0.5,1.0,2.0,3.0,4.0,5.0,6.0 mg·ml-1, and set 0 mg·ml-1 group as control. Proliferation curve of the cells was detected by the means of MTT method. For the cell immigration test, cells were treated with the medium with different concentrations of soybean agglutinin, i.e.0,0.5,1.0,1.5 mg·ml-1 and the number of immigration cells in different groups recorded.
After synchronizing the growth of the cells, the growth curve showed that cells grew very well. At about 12h after seeding, the cells got into a logarithmic phase, from 48h to 84h the cells were in a plateau phase and after that they got into a decline phase. The results of labeling showed that there were a lot of soybean agglutinin receptors on the cells of IEC-6.
The molecular weight of soya bean agglutinin was 30.78kDa, the level of purity was 98% and the value of agglutination was 1/210. All of these were not significantly different with those of Sigma standard soybean agglutinin (P>0.05). The values of agglutination of growth medium group and the control group were both 1/210. It indicated that the components of the cell growth medium had no influence on the agglutinative activity.
With time, results of the proliferation test revealed that the number of activity in all cell groups rose at first and then declined later. After adding the growth medium at 72h, cell activity approached the summit, but begun to decline thereafter. In 0.5 and 1.0 mg·ml-1 groups the activity of cells was significantly higher than the control group (P<0.05), while significantly lower in the 3.0 mg·ml-1 (P<0.05), and the 4.0,5.0, 6.0mg·ml-1 groups (P<0.01), respectively. This indicated that low concentration of soybean agglutinin could enhance the proliferation of the cells and that high concentration of SBA does not or even inhibit it. Cells immigrated from the cover-glasses in all groups with the number of immigration becoming higher with the increase of the soybean agglutinin concentration. At the highest concentration group, the numbers of cells numbers were significantly higher than all the other groups (P<0.05). Using this concentration of soybean agglutinin could enhance the immigration of the cell.
引文
[1]吴洁英等,建议在大豆制品的理化指标中增加脲酶定性试验,食品科学,1992,13(7)：48.
[2]Qin G. X 1996.Processing soybeans of different origins. Responses of a Chinese and a western pig breed to dietary inclusion:[dissertation]. Wageningen, The Netherland:Wageningen Agricultural University.
[3]Hartke, J. L., Apgar, G. A., Griswold, K. E., Jacobson, B. N., Rosenthal, T. L., Guthrie, T. A., 2003.Responses of weaning pigs to spray-dried animal plasma added to simple diets containing vary in levels of soya-bean meal:Animal Science Volume:77 Issue:1.
[4]Christopher T. Cordle. Soy Protein Allergy:Incidence and Relative Severity. Fifth International Symposiumon the Role of Soy in Preventing and Treating Chronic Disease. J. Nutr.134:1213S-1219S, May 2004.
[5]Pusztai, A., Ewen, S. W., Grant, G., et al. Relationship between survival and binding of plant lectins during small intestinal passage and their effectiveness as growth factors[J]. Digestion,1990,46 (12): 308-316.
[6]Li Z, Li D, and Qiao S. et al. Effects of soybean agglutinin on nitrogen metabolism and on characteristics of intestinal tissues and pancreas in rats[J]. Arch Tierernahr.2003,57(5):369-380.
[7]Greer F, Pusztai A. Toxicity of kidney bean (Phaseolus vulgaris) in rats:changes in intestinal permeability [J]. Digestion,1985,32:42(Abstr.).
[8]Gomes, E., Ortiz, V., Ventura, J., Campos, R. and Bourges, H. (1995) in:Advances in Mucosal Immunology (Mestecky, J. ed.)pp.533-536, Plenum, New York.
[9]Koninkx, J. F., Brown, D. S., Kok, W., et al. Polyamine metabolism of enterocyte-like Caco-2 cells after exposure to phaseolus vulgaris lectin[J]. Gut 3847-52.1996.
[10]汤树生.大豆凝集素与小肠的结合规律及其对大鼠免疫功能的影响[D].中国农业大学.2006.42(17)：10-13.
[11]王彬,胡海霞,孙青松等,大豆凝集素与肉鸡小肠粘膜细胞结合规律的研究[J].畜牧与饲料科学.2009,30(9)：15-16.
[12]L. G. Buttlea, A. C. Burrells. The binding of soybean agglutinin (SBA) to the intestinal epithelium of Atlantic salmon, Salyno solar and Rainbow trout, Oncorhynchus mykiss, fed high levels of soybean meal[J]. Veterinary Immunology and Immunopathology.2001,80:237-244.
[13]Quaroni A, Wands J, Trelstad RL, et al. Epithelial cell cultures from rat small intestine. JCell Biol.1979, 80:248-265.
[14]Franz, H. (1988) The ricin story. Adv. Lectin Res.,1,10-25.
[15]Landsteiner K, Raubitschek H. Be obachrungenuberHamolyseund Hamagglutination [J]. Zbl Bakt I Abt Orig,1907,45:600-607.
[16]Boyd, W. C., Reguera, R. M. Studies on haemagglutinins present in seeds of some representatives of the family leguminosae[J]. Immun.1949,62:333-339.
[17]Renkonnen, K. O. Studies on hemagglutinins present in the seeds of some representatives of the family leguminoseae[J]. Ann Med Exp Fen,1948,26(5):66-72.
[18]Goldstein, I. J., Winter, H. C., Poretz, R. D. Plant lectins:tools for the studying of complex carbohydrates. In Glycoproteins Ⅱ, (ed. J. Montreuil, Vliegenthart, J. F. G., Schacter, H. (Eds.)), pp. 403-174:Elsevier, Amsterdam.1997.
[19]Kocourek, J. V., Horejsi. Note on the recent discussion of the term lectin[J],1983,933-45.
[20]李德发.大豆抗营养因子[M].北京：中国科学技术出版社,2003：78.
[21]Van Damme, E. J. M., Peumans, W. J., Barre, A., et al. Plant lectins:a composite of several distinct families of structurally and evolutionary related proteins with diverse biological role[J]. Crit. Rev. Plant Sci. 1998.17:575-692.
[22]陈惠黎.糖复合物的结构和功能[M].上海医科大学出版社,(1997).
[23]余燕泽,石安静.贝类血细胞研究进展[J].动物学杂志,1998,33(5)：40-41.
[24]史久华.国外医学预防·诊断·治疗用生物制品分册,1995,18(3)：97.
[25]陈皓文,牟敦彩,李光友.微生物凝集紊的研究[J].自然杂志,2000,22(2)：95-100.
[26]Sharon, N & Lis. Lectines as cell recognition[J]. Molecules Science,1989.246-227.
[27]郭锰.凝集素的双功能性质：凝集素新定义[J].国外医学分子生物学分册,1990.12-35.
[28]Parkin, S., Rupp, B.& Hope, H. Atomic resolution structure of concanavalin A at 120 K, Acta Cryst. Sect.1996,52,1161-1168.
[29]Sharon, N., Lis, H. Legume lectins-a large family of homologous proteins[J]. FASEB.1990,4(3): 198-208.
[30]Weis, W. I. Drickamer, K. Structural basis of lectin-carbohydrate recognition[J], Annu. Rev. Biochem. 1996,65,441-473.
[31]Einspahr, H., E. H. Parks, and F. L. Suddath. The crystal structure of pea lectin at 3.0-A resolution[J]. Biol. Chem.1986,261:16518-16527.
[32]Dessen, A., D. Gupta, S. Sabesan, C. F. Brewer, and J. C. Sacchettini. X-ray crystal structure of the soybean agglutinin cross-linked with a biantennary analog of the blood group Ⅰ carbohydrate antigen[J], Biochemistry 1995,34:4933-4942.
[33]Hamelryck, T. W., M. H. Dao-Thi, F. Poortmans, M. J. Chrispeels, L. Wyns, and R. Loris. The crystallographic structure of phytohemagglutinin-L[J]Biol. Chem.1996,271:20479-20485
[34]Rao, V. S., Lam, K., Qa SBA, P. K. Three dimensional structure of the soybean agglutinin Gal/GalNAc complexes by homology modeling[J]. Biomol Struct Dyn.1998,15 (5):853-60.
[35]Thalles B. Grangeiro', Albert Schriefer, et al. Molecular cloning and characterization of ConBr, the lectin of Canavalia brasiliensis seeds[J]. Biochem.1997,248:43-48.
[36]Rao, V. S., Lam, K., QaSBA, P. K. Three dimensional structure of the soybean agglutinin Gal/GalNAc complexes by homology modeling[J]. JBiomol Struct Dyn,15 (5):853-60.1998.
[37]Rudiger, H. Chapman & Hall, Weinheim. Structure and function of plant lectins in Glycosciences. Status and perspectives,1997:415-438.
[38]Gatehouse, A. M. R., Powell, K. S., Peumans, W. J., Van Damme, E. J. M., atehouse, E. J. M. Insecticidal properties of plant lectins:their potential in plant protection. In:Pusztai, A., Bardocz, S. (Eds.), Lectins Biomedical Perspectives. Taylor Francis Ltd., London,1995,35-37.
[39]Mostafa M M. Chemical and urtritional changes in soybean during germination[J]. Food Chem.1987,23: 257-275.
[40]Gatehouse, A. M. R., Powell, K. S., Peumans, W. J., Van Damme, E. J. M., atehouse, E. J. M., 1995.Insecticidal properties of plant lectins:their potential in plant protection. In:Pusztai, A., Bardocz, S. (Eds.), Lectins Biomedical Perspectives. Taylor Francis Ltd., London, pp.35-37.
[41]王逸群,荆玉祥.豆科植物凝集素及其对根瘤菌的识别作用[J].植物学通报,2000,17(2)：127-132.
[42]Pieternel van Rhijn, Robert B. Goldberg, and Ann M. Hirsch et al. Lotus corniculatus Nodulation Specificity Is Changed by the Presence of a Soybean Lectin Gene[J]. The Plant Cell,1998,10:1233-1249.
[43]Cavada, B. S., Moreira, R. A,, Oliveira, J. T. A.& Grangeiro, T. B. Primary structures and functions of plant lectins[J]. Rev. Bras. Fisiol.1993,5:193-201.
[44]Prakash M. Dey, Surbhi Naik, John B. Pridham et al. The lectin nature of α-galactosidases from Vicia faba seeds [J]. FEBS Letters,1982,150(1):233-237.
[45]Maria Ines Isla, Marta Amelia Vattuone, Antonio Rodolfo Sampietro. et al. Proteinaceous inhibitor from Solanum tuberosum invertase[J]. Phytochemistry,1991,30(3):739-743.
[46]许兵等.麦胚凝集素促进对虾免疫功能的初步研究[J].青岛海洋大学学报,1992,22(4)：29.
[47]牟海津,江晓路,刘树青,管华诗.双壳贝类血清中凝集素凝集性能初步研究—对脊椎动物血细胞的凝集作用[J].青岛海洋大学学报,1999,29(2)：249-254.
[48]Rocio Coutino-Rodriguez, Pedro Hernandez-Cruz, Hector Giles-Rios ec tal. Lectins in Fruits Having Gastrointestinal Activity:Their Participation in the Hemagglutinating Property of Escherichia coli 0157:H7[J]. Archives of Medical Research.2001(32):251-257.
[49]陈皓文,孙丕喜,宋庆云.外源凝集素-水产动物御敌的有力兵器[J].黄渤海海洋,.1995,13(3)：61.
[50]周柔丽.哺乳动物凝集素及其生物学作用[J].生命的化学,1995,15(1)：12-15.
[51]江红.半乳糖凝集素的结构和功能[J].生命的化学,1995,15(6)：21-22.
[52]罗依惠等.草鱼血清凝集素的研究[J].浙江大学学报理学版,1999,26(2)：69-74.
[53]Brandley BK and Schmaar RL:Cell-surface carbohydrates in cell recognition and response [J]. Leukoc Biol,1986.40:97.
[54]周占祥,邓泽沛.羊精子在成熟和获能过程中表面凝集素标记变化[J].解剖学报,1994,25(3)：277-280.
[55]Malmi R and Sodertrom K-O.Lectin binding to carcinoma-in situ cells of the testis. A comparative study of CIS germ cells and seminoma cells[J]. Virehows Arch(A).1988,413:69-75.
[56]Rudiger, H. Chapman & Hall, Weinheim. Structure and function of plant lectins in Glycosciences. Status and perspectives,1997,415-438.
[57]Riidiger, H. (1997) Structure and function of plant lectins in Glycosciences. Status and perspectives (Gabius, H.-J.& Gabius, S., eds)pp.415-438, Chapman & Hall, Weinheim.
[58]Grant G, Watt W B, Stewared JC S, et al. Effects of dietary soy-bean lectin and trypsin inhibitors upon the pancreas of rat [J]. Med Sci Res,1987,15:1197-1198.
[59]Li, Zhentian, D. F. Li, and Shiyan Qiao.2003b. Effects of soybean agglutinin on nitrogen metabolism and on characterics of intestinal tissues and pancreas in rats. Arch. Anim. Nutr.,57(5):369-380.
[60]Salgado, P., Freire, J. P. B., Mourato, M., et al. Comparative effects of different legume protein sources in weaned piglets:nutrient digestibility, intestinal morphology and digestive enzymes[J].. Livest. Prod. Sci, 74191-202.2002.
[61]Hisayasu S, Orimo H, Migita S, Ikeda Y, Satoh K, Shinjo S, Hirai Y, Yoshino Y. et al. Soybean Protein Isolate and Soybean Lectin Inhibit Iron Absorption in Rats [J]. American Institute of Nutrition.1992,122(5): 1190-1196.
[62]T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[63]Carbonaro, M., Cappelloni, M., Nicoli, S., Lucarini, M., Carnovale, E., Solubility-digestibility relationship of legume proteins. Journal of Agricultural and Food Chemistry 1997,45:3387-3394.
[64]T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[65]Boris Tchernychev, Meir Wilchek. Natural human antibodies to dietary lectins[J]. Federation of European Biochemical Societies.1996,9(397):139-142.
[66]Y. O. Fasina, J. D. Garlich, H. L. Classen, et al. Response of Turkey Poults to Soybean Lectin Levels Typically Encountered in Commercial Diets.1.Effect on Growth and Nutrient Digestibility [J]. Poultry Science.2004,83:1559-1571.
[67]刘琳娜,不同种属动物对大豆凝集素敏感性的比较研究[D].吉林农业大学,2005.
[68]王幸,马秋枝,刘强等.大豆凝集素对小肠上皮细胞和红细胞凝集能力的研究[J].动物营养学报, 2008,20(2)：234-237.
[69]曾仲奎,吴洽庆.植物凝集素在植物体内的生理作用[J].天然产物研究与开发.1993.9.5(3)：69-76.
[70]冯佑民,王克夷,顾嘉绷,等。若干外源凝集素对胰岛素与其受体相互作用的影响[J].生物化学与生物物理学报,1981,9,13(4)：391-395.
[71]Gabius, H.-J.& Gabius, S. Glycusciences. StOpdenakker, G., Rudd, P. M., Ponting, C. P.& Dwek, R. A. Concepts and principles of glycobiology[J]. FASEB J.1993,7:1330-1337. atus and perspectives, Chapman & Hall, Weinheim.1997.
[72]Lis, H.& Sharon, N. Protein glycosylation. Structural and functional aspects, [J]. Biochem.1993,218: 1-27.
[73]Varki, A. Biological roles of oligosaccharides:all the theories are correct, Glycobiology[J].1993,3: 97-130.
[74]Gabius, H.-J. Animal lectins[J]. Biochem.1997),243,543-576.
[75]Opdenakker, G., Rudd, P. M., Ponting, C. P.& Dwek, R. A. Concepts and principles of glycobiology[J]. FASEB J.1993,7:1330-1337.
[76]Shur, B. D. The beginning of a sweet tale[J]. Curr: Bid.1994,4.996-999.
[77]Varki A. Biological roles of oligosaccharides:all of the theories are correct[J]. Glycobiology,1993,3(2): 97-130.
[78]Colin H R. Galectins asmodulators of cell adhesion [J]. Biochimie,2001,83:667-676.
[79]J. T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[80]Fiete D, Baenziger JU. Isolation of the SO4-4-GalNAc betal 4GlcNAc betal,2Man alpha-specific receptor from rat liver[J]. Biol Chem,1997,272(23):14629-14637.
[81]Fiete DJ, Beranek MC, Baenziger JU. A cysteine2rich domain of the"mannose receptor"mediates GalNAc-4-SO4 binding [J]. Proc Natl Acad Sci USA.1998,95(5):2089-2093.
[82]Reuben LOTAN, Rene CACAN, Monique CACAN, et al. On the presence of two types of subunit in toybean agglutinin[J]. North-Holland Publishing Company.1975,9,57(1):100-103.
[83]Liener, I. E., M. J. Pallansch. Purification of a toxic substance from the defatted soybean flour [J]. Biol. Chem.1952,197:29-36.
[84]Charles L. J, Sarah Ehrlich-Rogozinski, Halina Lis et al. Transition metal requirements of soybean agglutinin[J]. Febs Letters.1977,82(2):191-196.
[85]王忠艳,白秀娟.热处理条件参数对豆粕中大豆凝集素活性的影响[J].野生动物杂志.2008,29(6)：305-307.
[86]戴大章,陈妙月,刘建新等.大豆凝集素的灭活研究[J].粮油加工与食品机械.2003,5：45-47.
[87]Dessen, A., Gupta, D., Sabesan, S., et al. X-ray crystal structure of the soybean agglutinin cross-linked with a biantennary analog of the blood group I carbohydrate antigen[J]. Biochemistry,34 (15):4933-42.1995.
[88]马秋枝,大豆凝集素糖配体的筛选及其对体内结合率的影响[南京农业大学硕土论文].南京农业大学,2007.
[89]DeBoeck H, Lis H, van TilbeurghH, Sharon N, Loonteins F G Bingding of simple carbodydarte and some chromoPhored derivatives of soybean agglutinin as follows by titrimetric procedured and stopped-flow kineteds[J]. Biol. Chem.1984,259:7067-7074.
[90]Sharon, N., Lis, H. How proteins bind carbohydrates:lessons from legume lectins[J]. Agric Food Chem. 2002,50(22):6586-6591.
[91]Pusstai, A., B. Willam, J. C. Stewart. A comprehensive scheme for the isolation of trypsin inhibitors and the agglutinin from soybean seeds[J]. Agric. Food Chem.1991,39:862-866.
[92]Krugluger, W., T. Lucas., M. Koller et al. Soybean agglutinin binds a 160-kD a rat macrophage membrane glycoprotein and enhances cell diferentiation and activation[J]. Immunol. Lett.1996,52:53-56.
[93]Zhu, Q. L., J. Meisinger, D. H. VanThiel et al. Efects of soybean extract onmorphology and survival of Caco-2, SW620, andHT-29cells[J]. Nutr. Cancer2002.42:131.
[94]de Oliveira JT A, Pusztai A, Grant G.1998. Changes in organs and tissues induced by feeding of purified kidney (phaseolus vulgaris) lectin. Nutri. Res.,8:943-947.
[95]Li, Zhentian, D. F. Li, Shiyan Qiao and Changhai Huang.2003a. Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat. Arch. Anim. Nutr.,57(4):267-277.
[96]Makind M O, Umapathy E, Akingbem B T I, Manisodza K T, Skadhasuge E.1996.Effects of dietary soybean and cowpea on gut morphology and faecal composition in creep and non creep fed pigs. J. Vet. Med., Ser. A,43:75-85.
[97]Douglas M W, Parsons C M, Hymowitz T.1999. Nutritional evaluation of lectin free soybeans for poultry. Poult. Sci.,78:91-95.
[98]Sborne, T. B., and L. B. Mendel. The use of soy bean as food. J. Biol. Chem.1917,32:369-387.
[99]Liener, I. E. Non-nutritive factors and bioactive compounds in soy in Animal Nutrition. J. K. Drackley, ed. Fed. Anim. Sci. Soc., Savoy, IL.2000.13-45.
[100]Douglas, M. W., Parsons, C. M., Hymowitz, T. et al. Nutritional evaluation of lectin-free soybeans for poultry[J]. Poult Sci,78 (1):91-5.1999.
[101]Liener, I. E. Effects of processing on antinutritional factors in legumes:the soybean case[J]. Arch Latinoam Nutr,1996,44(4 Suppl 1):48S-54S.
[102]Michele W. Douglas, Carl M. Parsons, and Theodore Hymowitz. et al. Nutritional Evaluation of Lectin-Free Soybeans for Poultry[J]. Poultry Science.1999,78:91-95.
[103]M. F. Palacios, R. A. Easter, and K. T. Soltwedel et al. Effect of soybean variety and processing on growth performance of young chicks and pigs[J]. Anim Sci.2004,82:1108-1114.
[104]李振田.大豆凝集素的检测、纯化和对大鼠抗营养机理的研究[D].中国农业大学,2003.
[105]Rouanet, J. M., Besancon, P., Lafont, J. Effect of lectins from leguminous seeds on rat duodenal enterokinase activity[J]. Experientia,1983,39 (12):1356-8.
[106]臧建军,马永喜,朴香淑,等.大豆凝集素抗营养作用的研究进展[J].中国饲料,(06).2008.
[107]Fasina, Y. O., Garlich, J. D., Classen, H. L., et al. Response of turkey poults to soybean lectin levels typically encountered in commercial diets.1.Effect on growth and nutrient digestibility [J]. Poult Sci,83 (9):1559-71.2004.
[108]Douglas M, W, Parsons C M, Hymowitz T. Nutritional evaluation of lectin-free soybeans for poultry [J]. Point. Sci,1999,78:91-95.
[109]Lis, H. and Sharon, N. in:The Lectins:Properties, Functions and Applications in Biology and Medicine (Liener, I. E., Sharon, N. and Goldstein, I. J. eds.)Academic Press, pp. (1986) 293-370.
[110]Orlando, FL. Brooks SA, Dwek MV, Schumacher U. Functional and molecular glycobiology. Oxford: BIOS Scientific Publishers Limited,2002.
[111]Arya M. Vanha-Perttula T. Leciin binding pattern of bull testis and epididymis[J]Androl.11985,6(4): 230-242.
[112]Magargee S F et al. Changes in lectin-binding features of ram sperm surfaces associated with epididymal maturation and ejaculation[J]. Biol Reprod,1988,38:667-685.
[113]Magargee S F et al. Changes in lectin-binding features of ram sperm surfaces associated with epididymal maturation and ejaculation[J]. Biol Reprod,1988,38:667-685.
[114]王一飞等.外源性凝集素引起人精子凝集反应的定量观察[J].生殖与避孕.1984,38：667-685.
[115]Malmi R and Sodertrom K-O.Lectin binding to carcinoma-in situ cells of the testis. A comparative study of CIS germ cells and seminoma cells[J]. Virehows Arch(A).1988,413:69-75.
[116]Ronald A. Advances in immunohistochemistry, new York:Mosson Dub USA Ine.1984:67-63.
[117]Lis, H.& Sharon, N. Lectins as molecules and as tools, Annu. Rev. Biochem.1986,55:35-61.
[118]Wasano K, Hirakawa Y. Recombinant galectin 21 recognizes mucin and epithelial cell surface glycocalyces of gastrointestinal tract [J]. Histochem Cytochem.1997,45:275-283.
[119]苏秉忠,高中明.胃和十二指肠粘膜常见病变大豆凝集素受体分布的研究[J].内蒙古医学院学报。1990.12(1)：45-48.
[120]Pusztai, A. Biological effects of dietary lectins. In Recent advances of research in antinutritional factors in legume seeds, (eds. J. Huisman A. F. B. Van der poel and I. E. Liener), pp.17-29. The Netherlands:Pudoc Wageningen.1989.
[121]Krugluger, W., T. Lucas., M. Koller et al. Soybean agglutinin binds a 160-kD a rat macrophage membrane glycoprotein and enhances cell diferentiation and activation[J]. Immunol. Lett.1996,52:53-56.
[122]任锡麟,林钧材,顾天爵等.正常小鼠干细胞和腹水性肝癌细胞对的大豆凝集素受体研究[J].肿瘤,1982.9(848)：4-8.
[123]Quaroni A, Wands J, Trelstad RL, et al. Epithelial cell cultures from rat small intestine. JCell Biol.1979, 80:248-265.
[124]J. T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[125]Krugluger, W., T. Lucas., M. Koller et al. Soybean agglutinin binds a 160-kD a rat macrophage membrane glycoprotein and enhances cell diferentiation and activation[J]. Immunol. Lett.1996,52:53-56.
[126]Jaffe, W. G. Haemaggutinins. In Toxic Constituents in Plant Food stuffs[M]. Academic Press,1980,7(1): 80.
[127]Liener I E. The photometric determination of the hameagglutination activity soyin and crude soya bean extracts [J]. Archives of Bio-chemistry and Biophysics,1955,54:223-226.
[128]Grant G, More L J, McKenzie N H, et al. A survey of the nutritional and haeagglutinin properties of legume seeds generally available in the UK [J]. British Journal of Nutrition,1983,50:207-212.
[129]Van Schukthass G K, Giglio Me ConnellD S, Benedek GB. Detection of agglutinination reactions using anisotrop ic light scattering:an immunoassay of high sensitivity [J]. Molecular Immunology,1980,17:81-82.
[130]Kohle H, Kauss H. Imp roved analysis of haemagglutination assays for quantination of lectin activity [J]. Analytical Biochemistry,1980,103:227-229.
[131]Thalles B. GRANGEIRO', Albert SCHRIEFER'et al. Molecular cloning and characterization of ConBr, the lectin of Canavalia brasiliensis seeds[J]. Biochem.1997,248,43-48.
[132]Douglas J. Turner, MD, Paul C. Martin, et al. Substance P Regulates Migration in Rat Intestinal Epithelial Cells[J]. Annals of Surgery.2007,3,245(3):408-414.
[133]Guo X, Rao JN, Liu L, et al. Polyamines regulate catenin tyrosine phosphorylation via Ca2+ during intestinal epithelial cell migration[J]. Am JPhysiol Cell Physiol.2002,283:C722-C734.
[134]冯琳.大豆凝集素对鲤鱼肠道上皮细胞增殖分化及其功能的影响[四川农业大学学位论文].四川农业大学,2006.
[135]Zhu, Q. L., J. Meisinger, D. H. VanThiel et al. Efects of soybean extract onmorphology and survival of Caco-2, SW620, andHT-29cells[J]. Nutr. Cancer2002.42:131.
[2]Qin G. X 1996.Processing soybeans of different origins. Responses of a Chinese and a western pig breed to dietary inclusion:[dissertation]. Wageningen, The Netherland:Wageningen Agricultural University.
[3]Hartke, J. L., Apgar, G. A., Griswold, K. E., Jacobson, B. N., Rosenthal, T. L., Guthrie, T. A., 2003.Responses of weaning pigs to spray-dried animal plasma added to simple diets containing vary in levels of soya-bean meal:Animal Science Volume:77 Issue:1.
[4]Christopher T. Cordle. Soy Protein Allergy:Incidence and Relative Severity. Fifth International Symposiumon the Role of Soy in Preventing and Treating Chronic Disease. J. Nutr.134:1213S-1219S, May 2004.
[5]Pusztai, A., Ewen, S. W., Grant, G., et al. Relationship between survival and binding of plant lectins during small intestinal passage and their effectiveness as growth factors[J]. Digestion,1990,46 (12): 308-316.
[6]Li Z, Li D, and Qiao S. et al. Effects of soybean agglutinin on nitrogen metabolism and on characteristics of intestinal tissues and pancreas in rats[J]. Arch Tierernahr.2003,57(5):369-380.
[7]Greer F, Pusztai A. Toxicity of kidney bean (Phaseolus vulgaris) in rats:changes in intestinal permeability [J]. Digestion,1985,32:42(Abstr.).
[8]Gomes, E., Ortiz, V., Ventura, J., Campos, R. and Bourges, H. (1995) in:Advances in Mucosal Immunology (Mestecky, J. ed.)pp.533-536, Plenum, New York.
[9]Koninkx, J. F., Brown, D. S., Kok, W., et al. Polyamine metabolism of enterocyte-like Caco-2 cells after exposure to phaseolus vulgaris lectin[J]. Gut 3847-52.1996.
[10]汤树生.大豆凝集素与小肠的结合规律及其对大鼠免疫功能的影响[D].中国农业大学.2006.42(17)：10-13.
[11]王彬,胡海霞,孙青松等,大豆凝集素与肉鸡小肠粘膜细胞结合规律的研究[J].畜牧与饲料科学.2009,30(9)：15-16.
[12]L. G. Buttlea, A. C. Burrells. The binding of soybean agglutinin (SBA) to the intestinal epithelium of Atlantic salmon, Salyno solar and Rainbow trout, Oncorhynchus mykiss, fed high levels of soybean meal[J]. Veterinary Immunology and Immunopathology.2001,80:237-244.
[13]Quaroni A, Wands J, Trelstad RL, et al. Epithelial cell cultures from rat small intestine. JCell Biol.1979, 80:248-265.
[14]Franz, H. (1988) The ricin story. Adv. Lectin Res.,1,10-25.
[15]Landsteiner K, Raubitschek H. Be obachrungenuberHamolyseund Hamagglutination [J]. Zbl Bakt I Abt Orig,1907,45:600-607.
[16]Boyd, W. C., Reguera, R. M. Studies on haemagglutinins present in seeds of some representatives of the family leguminosae[J]. Immun.1949,62:333-339.
[17]Renkonnen, K. O. Studies on hemagglutinins present in the seeds of some representatives of the family leguminoseae[J]. Ann Med Exp Fen,1948,26(5):66-72.
[18]Goldstein, I. J., Winter, H. C., Poretz, R. D. Plant lectins:tools for the studying of complex carbohydrates. In Glycoproteins Ⅱ, (ed. J. Montreuil, Vliegenthart, J. F. G., Schacter, H. (Eds.)), pp. 403-174:Elsevier, Amsterdam.1997.
[19]Kocourek, J. V., Horejsi. Note on the recent discussion of the term lectin[J],1983,933-45.
[20]李德发.大豆抗营养因子[M].北京：中国科学技术出版社,2003：78.
[21]Van Damme, E. J. M., Peumans, W. J., Barre, A., et al. Plant lectins:a composite of several distinct families of structurally and evolutionary related proteins with diverse biological role[J]. Crit. Rev. Plant Sci. 1998.17:575-692.
[22]陈惠黎.糖复合物的结构和功能[M].上海医科大学出版社,(1997).
[23]余燕泽,石安静.贝类血细胞研究进展[J].动物学杂志,1998,33(5)：40-41.
[24]史久华.国外医学预防·诊断·治疗用生物制品分册,1995,18(3)：97.
[25]陈皓文,牟敦彩,李光友.微生物凝集紊的研究[J].自然杂志,2000,22(2)：95-100.
[26]Sharon, N & Lis. Lectines as cell recognition[J]. Molecules Science,1989.246-227.
[27]郭锰.凝集素的双功能性质：凝集素新定义[J].国外医学分子生物学分册,1990.12-35.
[28]Parkin, S., Rupp, B.& Hope, H. Atomic resolution structure of concanavalin A at 120 K, Acta Cryst. Sect.1996,52,1161-1168.
[29]Sharon, N., Lis, H. Legume lectins-a large family of homologous proteins[J]. FASEB.1990,4(3): 198-208.
[30]Weis, W. I. Drickamer, K. Structural basis of lectin-carbohydrate recognition[J], Annu. Rev. Biochem. 1996,65,441-473.
[31]Einspahr, H., E. H. Parks, and F. L. Suddath. The crystal structure of pea lectin at 3.0-A resolution[J]. Biol. Chem.1986,261:16518-16527.
[32]Dessen, A., D. Gupta, S. Sabesan, C. F. Brewer, and J. C. Sacchettini. X-ray crystal structure of the soybean agglutinin cross-linked with a biantennary analog of the blood group Ⅰ carbohydrate antigen[J], Biochemistry 1995,34:4933-4942.
[33]Hamelryck, T. W., M. H. Dao-Thi, F. Poortmans, M. J. Chrispeels, L. Wyns, and R. Loris. The crystallographic structure of phytohemagglutinin-L[J]Biol. Chem.1996,271:20479-20485
[34]Rao, V. S., Lam, K., Qa SBA, P. K. Three dimensional structure of the soybean agglutinin Gal/GalNAc complexes by homology modeling[J]. Biomol Struct Dyn.1998,15 (5):853-60.
[35]Thalles B. Grangeiro', Albert Schriefer, et al. Molecular cloning and characterization of ConBr, the lectin of Canavalia brasiliensis seeds[J]. Biochem.1997,248:43-48.
[36]Rao, V. S., Lam, K., QaSBA, P. K. Three dimensional structure of the soybean agglutinin Gal/GalNAc complexes by homology modeling[J]. JBiomol Struct Dyn,15 (5):853-60.1998.
[37]Rudiger, H. Chapman & Hall, Weinheim. Structure and function of plant lectins in Glycosciences. Status and perspectives,1997:415-438.
[38]Gatehouse, A. M. R., Powell, K. S., Peumans, W. J., Van Damme, E. J. M., atehouse, E. J. M. Insecticidal properties of plant lectins:their potential in plant protection. In:Pusztai, A., Bardocz, S. (Eds.), Lectins Biomedical Perspectives. Taylor Francis Ltd., London,1995,35-37.
[39]Mostafa M M. Chemical and urtritional changes in soybean during germination[J]. Food Chem.1987,23: 257-275.
[40]Gatehouse, A. M. R., Powell, K. S., Peumans, W. J., Van Damme, E. J. M., atehouse, E. J. M., 1995.Insecticidal properties of plant lectins:their potential in plant protection. In:Pusztai, A., Bardocz, S. (Eds.), Lectins Biomedical Perspectives. Taylor Francis Ltd., London, pp.35-37.
[41]王逸群,荆玉祥.豆科植物凝集素及其对根瘤菌的识别作用[J].植物学通报,2000,17(2)：127-132.
[42]Pieternel van Rhijn, Robert B. Goldberg, and Ann M. Hirsch et al. Lotus corniculatus Nodulation Specificity Is Changed by the Presence of a Soybean Lectin Gene[J]. The Plant Cell,1998,10:1233-1249.
[43]Cavada, B. S., Moreira, R. A,, Oliveira, J. T. A.& Grangeiro, T. B. Primary structures and functions of plant lectins[J]. Rev. Bras. Fisiol.1993,5:193-201.
[44]Prakash M. Dey, Surbhi Naik, John B. Pridham et al. The lectin nature of α-galactosidases from Vicia faba seeds [J]. FEBS Letters,1982,150(1):233-237.
[45]Maria Ines Isla, Marta Amelia Vattuone, Antonio Rodolfo Sampietro. et al. Proteinaceous inhibitor from Solanum tuberosum invertase[J]. Phytochemistry,1991,30(3):739-743.
[46]许兵等.麦胚凝集素促进对虾免疫功能的初步研究[J].青岛海洋大学学报,1992,22(4)：29.
[47]牟海津,江晓路,刘树青,管华诗.双壳贝类血清中凝集素凝集性能初步研究—对脊椎动物血细胞的凝集作用[J].青岛海洋大学学报,1999,29(2)：249-254.
[48]Rocio Coutino-Rodriguez, Pedro Hernandez-Cruz, Hector Giles-Rios ec tal. Lectins in Fruits Having Gastrointestinal Activity:Their Participation in the Hemagglutinating Property of Escherichia coli 0157:H7[J]. Archives of Medical Research.2001(32):251-257.
[49]陈皓文,孙丕喜,宋庆云.外源凝集素-水产动物御敌的有力兵器[J].黄渤海海洋,.1995,13(3)：61.
[50]周柔丽.哺乳动物凝集素及其生物学作用[J].生命的化学,1995,15(1)：12-15.
[51]江红.半乳糖凝集素的结构和功能[J].生命的化学,1995,15(6)：21-22.
[52]罗依惠等.草鱼血清凝集素的研究[J].浙江大学学报理学版,1999,26(2)：69-74.
[53]Brandley BK and Schmaar RL:Cell-surface carbohydrates in cell recognition and response [J]. Leukoc Biol,1986.40:97.
[54]周占祥,邓泽沛.羊精子在成熟和获能过程中表面凝集素标记变化[J].解剖学报,1994,25(3)：277-280.
[55]Malmi R and Sodertrom K-O.Lectin binding to carcinoma-in situ cells of the testis. A comparative study of CIS germ cells and seminoma cells[J]. Virehows Arch(A).1988,413:69-75.
[56]Rudiger, H. Chapman & Hall, Weinheim. Structure and function of plant lectins in Glycosciences. Status and perspectives,1997,415-438.
[57]Riidiger, H. (1997) Structure and function of plant lectins in Glycosciences. Status and perspectives (Gabius, H.-J.& Gabius, S., eds)pp.415-438, Chapman & Hall, Weinheim.
[58]Grant G, Watt W B, Stewared JC S, et al. Effects of dietary soy-bean lectin and trypsin inhibitors upon the pancreas of rat [J]. Med Sci Res,1987,15:1197-1198.
[59]Li, Zhentian, D. F. Li, and Shiyan Qiao.2003b. Effects of soybean agglutinin on nitrogen metabolism and on characterics of intestinal tissues and pancreas in rats. Arch. Anim. Nutr.,57(5):369-380.
[60]Salgado, P., Freire, J. P. B., Mourato, M., et al. Comparative effects of different legume protein sources in weaned piglets:nutrient digestibility, intestinal morphology and digestive enzymes[J].. Livest. Prod. Sci, 74191-202.2002.
[61]Hisayasu S, Orimo H, Migita S, Ikeda Y, Satoh K, Shinjo S, Hirai Y, Yoshino Y. et al. Soybean Protein Isolate and Soybean Lectin Inhibit Iron Absorption in Rats [J]. American Institute of Nutrition.1992,122(5): 1190-1196.
[62]T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[63]Carbonaro, M., Cappelloni, M., Nicoli, S., Lucarini, M., Carnovale, E., Solubility-digestibility relationship of legume proteins. Journal of Agricultural and Food Chemistry 1997,45:3387-3394.
[64]T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[65]Boris Tchernychev, Meir Wilchek. Natural human antibodies to dietary lectins[J]. Federation of European Biochemical Societies.1996,9(397):139-142.
[66]Y. O. Fasina, J. D. Garlich, H. L. Classen, et al. Response of Turkey Poults to Soybean Lectin Levels Typically Encountered in Commercial Diets.1.Effect on Growth and Nutrient Digestibility [J]. Poultry Science.2004,83:1559-1571.
[67]刘琳娜,不同种属动物对大豆凝集素敏感性的比较研究[D].吉林农业大学,2005.
[68]王幸,马秋枝,刘强等.大豆凝集素对小肠上皮细胞和红细胞凝集能力的研究[J].动物营养学报, 2008,20(2)：234-237.
[69]曾仲奎,吴洽庆.植物凝集素在植物体内的生理作用[J].天然产物研究与开发.1993.9.5(3)：69-76.
[70]冯佑民,王克夷,顾嘉绷,等。若干外源凝集素对胰岛素与其受体相互作用的影响[J].生物化学与生物物理学报,1981,9,13(4)：391-395.
[71]Gabius, H.-J.& Gabius, S. Glycusciences. StOpdenakker, G., Rudd, P. M., Ponting, C. P.& Dwek, R. A. Concepts and principles of glycobiology[J]. FASEB J.1993,7:1330-1337. atus and perspectives, Chapman & Hall, Weinheim.1997.
[72]Lis, H.& Sharon, N. Protein glycosylation. Structural and functional aspects, [J]. Biochem.1993,218: 1-27.
[73]Varki, A. Biological roles of oligosaccharides:all the theories are correct, Glycobiology[J].1993,3: 97-130.
[74]Gabius, H.-J. Animal lectins[J]. Biochem.1997),243,543-576.
[75]Opdenakker, G., Rudd, P. M., Ponting, C. P.& Dwek, R. A. Concepts and principles of glycobiology[J]. FASEB J.1993,7:1330-1337.
[76]Shur, B. D. The beginning of a sweet tale[J]. Curr: Bid.1994,4.996-999.
[77]Varki A. Biological roles of oligosaccharides:all of the theories are correct[J]. Glycobiology,1993,3(2): 97-130.
[78]Colin H R. Galectins asmodulators of cell adhesion [J]. Biochimie,2001,83:667-676.
[79]J. T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[80]Fiete D, Baenziger JU. Isolation of the SO4-4-GalNAc betal 4GlcNAc betal,2Man alpha-specific receptor from rat liver[J]. Biol Chem,1997,272(23):14629-14637.
[81]Fiete DJ, Beranek MC, Baenziger JU. A cysteine2rich domain of the"mannose receptor"mediates GalNAc-4-SO4 binding [J]. Proc Natl Acad Sci USA.1998,95(5):2089-2093.
[82]Reuben LOTAN, Rene CACAN, Monique CACAN, et al. On the presence of two types of subunit in toybean agglutinin[J]. North-Holland Publishing Company.1975,9,57(1):100-103.
[83]Liener, I. E., M. J. Pallansch. Purification of a toxic substance from the defatted soybean flour [J]. Biol. Chem.1952,197:29-36.
[84]Charles L. J, Sarah Ehrlich-Rogozinski, Halina Lis et al. Transition metal requirements of soybean agglutinin[J]. Febs Letters.1977,82(2):191-196.
[85]王忠艳,白秀娟.热处理条件参数对豆粕中大豆凝集素活性的影响[J].野生动物杂志.2008,29(6)：305-307.
[86]戴大章,陈妙月,刘建新等.大豆凝集素的灭活研究[J].粮油加工与食品机械.2003,5：45-47.
[87]Dessen, A., Gupta, D., Sabesan, S., et al. X-ray crystal structure of the soybean agglutinin cross-linked with a biantennary analog of the blood group I carbohydrate antigen[J]. Biochemistry,34 (15):4933-42.1995.
[88]马秋枝,大豆凝集素糖配体的筛选及其对体内结合率的影响[南京农业大学硕土论文].南京农业大学,2007.
[89]DeBoeck H, Lis H, van TilbeurghH, Sharon N, Loonteins F G Bingding of simple carbodydarte and some chromoPhored derivatives of soybean agglutinin as follows by titrimetric procedured and stopped-flow kineteds[J]. Biol. Chem.1984,259:7067-7074.
[90]Sharon, N., Lis, H. How proteins bind carbohydrates:lessons from legume lectins[J]. Agric Food Chem. 2002,50(22):6586-6591.
[91]Pusstai, A., B. Willam, J. C. Stewart. A comprehensive scheme for the isolation of trypsin inhibitors and the agglutinin from soybean seeds[J]. Agric. Food Chem.1991,39:862-866.
[92]Krugluger, W., T. Lucas., M. Koller et al. Soybean agglutinin binds a 160-kD a rat macrophage membrane glycoprotein and enhances cell diferentiation and activation[J]. Immunol. Lett.1996,52:53-56.
[93]Zhu, Q. L., J. Meisinger, D. H. VanThiel et al. Efects of soybean extract onmorphology and survival of Caco-2, SW620, andHT-29cells[J]. Nutr. Cancer2002.42:131.
[94]de Oliveira JT A, Pusztai A, Grant G.1998. Changes in organs and tissues induced by feeding of purified kidney (phaseolus vulgaris) lectin. Nutri. Res.,8:943-947.
[95]Li, Zhentian, D. F. Li, Shiyan Qiao and Changhai Huang.2003a. Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat. Arch. Anim. Nutr.,57(4):267-277.
[96]Makind M O, Umapathy E, Akingbem B T I, Manisodza K T, Skadhasuge E.1996.Effects of dietary soybean and cowpea on gut morphology and faecal composition in creep and non creep fed pigs. J. Vet. Med., Ser. A,43:75-85.
[97]Douglas M W, Parsons C M, Hymowitz T.1999. Nutritional evaluation of lectin free soybeans for poultry. Poult. Sci.,78:91-95.
[98]Sborne, T. B., and L. B. Mendel. The use of soy bean as food. J. Biol. Chem.1917,32:369-387.
[99]Liener, I. E. Non-nutritive factors and bioactive compounds in soy in Animal Nutrition. J. K. Drackley, ed. Fed. Anim. Sci. Soc., Savoy, IL.2000.13-45.
[100]Douglas, M. W., Parsons, C. M., Hymowitz, T. et al. Nutritional evaluation of lectin-free soybeans for poultry[J]. Poult Sci,78 (1):91-5.1999.
[101]Liener, I. E. Effects of processing on antinutritional factors in legumes:the soybean case[J]. Arch Latinoam Nutr,1996,44(4 Suppl 1):48S-54S.
[102]Michele W. Douglas, Carl M. Parsons, and Theodore Hymowitz. et al. Nutritional Evaluation of Lectin-Free Soybeans for Poultry[J]. Poultry Science.1999,78:91-95.
[103]M. F. Palacios, R. A. Easter, and K. T. Soltwedel et al. Effect of soybean variety and processing on growth performance of young chicks and pigs[J]. Anim Sci.2004,82:1108-1114.
[104]李振田.大豆凝集素的检测、纯化和对大鼠抗营养机理的研究[D].中国农业大学,2003.
[105]Rouanet, J. M., Besancon, P., Lafont, J. Effect of lectins from leguminous seeds on rat duodenal enterokinase activity[J]. Experientia,1983,39 (12):1356-8.
[106]臧建军,马永喜,朴香淑,等.大豆凝集素抗营养作用的研究进展[J].中国饲料,(06).2008.
[107]Fasina, Y. O., Garlich, J. D., Classen, H. L., et al. Response of turkey poults to soybean lectin levels typically encountered in commercial diets.1.Effect on growth and nutrient digestibility [J]. Poult Sci,83 (9):1559-71.2004.
[108]Douglas M, W, Parsons C M, Hymowitz T. Nutritional evaluation of lectin-free soybeans for poultry [J]. Point. Sci,1999,78:91-95.
[109]Lis, H. and Sharon, N. in:The Lectins:Properties, Functions and Applications in Biology and Medicine (Liener, I. E., Sharon, N. and Goldstein, I. J. eds.)Academic Press, pp. (1986) 293-370.
[110]Orlando, FL. Brooks SA, Dwek MV, Schumacher U. Functional and molecular glycobiology. Oxford: BIOS Scientific Publishers Limited,2002.
[111]Arya M. Vanha-Perttula T. Leciin binding pattern of bull testis and epididymis[J]Androl.11985,6(4): 230-242.
[112]Magargee S F et al. Changes in lectin-binding features of ram sperm surfaces associated with epididymal maturation and ejaculation[J]. Biol Reprod,1988,38:667-685.
[113]Magargee S F et al. Changes in lectin-binding features of ram sperm surfaces associated with epididymal maturation and ejaculation[J]. Biol Reprod,1988,38:667-685.
[114]王一飞等.外源性凝集素引起人精子凝集反应的定量观察[J].生殖与避孕.1984,38：667-685.
[115]Malmi R and Sodertrom K-O.Lectin binding to carcinoma-in situ cells of the testis. A comparative study of CIS germ cells and seminoma cells[J]. Virehows Arch(A).1988,413:69-75.
[116]Ronald A. Advances in immunohistochemistry, new York:Mosson Dub USA Ine.1984:67-63.
[117]Lis, H.& Sharon, N. Lectins as molecules and as tools, Annu. Rev. Biochem.1986,55:35-61.
[118]Wasano K, Hirakawa Y. Recombinant galectin 21 recognizes mucin and epithelial cell surface glycocalyces of gastrointestinal tract [J]. Histochem Cytochem.1997,45:275-283.
[119]苏秉忠,高中明.胃和十二指肠粘膜常见病变大豆凝集素受体分布的研究[J].内蒙古医学院学报。1990.12(1)：45-48.
[120]Pusztai, A. Biological effects of dietary lectins. In Recent advances of research in antinutritional factors in legume seeds, (eds. J. Huisman A. F. B. Van der poel and I. E. Liener), pp.17-29. The Netherlands:Pudoc Wageningen.1989.
[121]Krugluger, W., T. Lucas., M. Koller et al. Soybean agglutinin binds a 160-kD a rat macrophage membrane glycoprotein and enhances cell diferentiation and activation[J]. Immunol. Lett.1996,52:53-56.
[122]任锡麟,林钧材,顾天爵等.正常小鼠干细胞和腹水性肝癌细胞对的大豆凝集素受体研究[J].肿瘤,1982.9(848)：4-8.
[123]Quaroni A, Wands J, Trelstad RL, et al. Epithelial cell cultures from rat small intestine. JCell Biol.1979, 80:248-265.
[124]J. T. A. Oliveira, F. J. B. Rios, and I. M. Vasconcelos et al. Cratylia argentea seed lectin, a possible defensive protein against plant-eating organisms:effects on rat metabolism and gut histology[J]. Food and Chemical Toxicology 2004,42:1737-1747.
[125]Krugluger, W., T. Lucas., M. Koller et al. Soybean agglutinin binds a 160-kD a rat macrophage membrane glycoprotein and enhances cell diferentiation and activation[J]. Immunol. Lett.1996,52:53-56.
[126]Jaffe, W. G. Haemaggutinins. In Toxic Constituents in Plant Food stuffs[M]. Academic Press,1980,7(1): 80.
[127]Liener I E. The photometric determination of the hameagglutination activity soyin and crude soya bean extracts [J]. Archives of Bio-chemistry and Biophysics,1955,54:223-226.
[128]Grant G, More L J, McKenzie N H, et al. A survey of the nutritional and haeagglutinin properties of legume seeds generally available in the UK [J]. British Journal of Nutrition,1983,50:207-212.
[129]Van Schukthass G K, Giglio Me ConnellD S, Benedek GB. Detection of agglutinination reactions using anisotrop ic light scattering:an immunoassay of high sensitivity [J]. Molecular Immunology,1980,17:81-82.
[130]Kohle H, Kauss H. Imp roved analysis of haemagglutination assays for quantination of lectin activity [J]. Analytical Biochemistry,1980,103:227-229.
[131]Thalles B. GRANGEIRO', Albert SCHRIEFER'et al. Molecular cloning and characterization of ConBr, the lectin of Canavalia brasiliensis seeds[J]. Biochem.1997,248,43-48.
[132]Douglas J. Turner, MD, Paul C. Martin, et al. Substance P Regulates Migration in Rat Intestinal Epithelial Cells[J]. Annals of Surgery.2007,3,245(3):408-414.
[133]Guo X, Rao JN, Liu L, et al. Polyamines regulate catenin tyrosine phosphorylation via Ca2+ during intestinal epithelial cell migration[J]. Am JPhysiol Cell Physiol.2002,283:C722-C734.
[134]冯琳.大豆凝集素对鲤鱼肠道上皮细胞增殖分化及其功能的影响[四川农业大学学位论文].四川农业大学,2006.
[135]Zhu, Q. L., J. Meisinger, D. H. VanThiel et al. Efects of soybean extract onmorphology and survival of Caco-2, SW620, andHT-29cells[J]. Nutr. Cancer2002.42:131.