远东拟沙丁鱼低聚肽化学组成及其增强免疫力功能评价
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摘要
为了评价远东拟沙丁鱼低聚肽(Sardinops sagax oligopeptide,SSO)增强免疫力的功能。以远东拟沙丁鱼为原料,通过酶解、离心、膜分离、喷雾干燥等工序制备低聚肽,分析其化学组成。通过动物实验,运用保健食品功能成分评价技术,对其增强免疫力功能实验进行研究。结果表明,远东拟沙丁鱼低聚肽分子质量主要在1 000 u以下,占总干重的66.4%,谷氨酸、赖氨酸、天冬氨酸与亮氨酸氨基酸含量较高,且含有丰富的硒元素,含量为1.66 mg/100g。与阴性对照组相比,阳性对照组、低聚肽高剂量组能显著提高正常小鼠脾淋巴细胞增殖能力(p<0.05),细胞免疫功能结果为阳性;阳性对照组、低聚肽高、中、低3个剂量组均能显著提高正常小鼠脾脏指数和空斑数(p<0.05),体液免疫功能结果为阳性;阳性对照组、低聚肽高、中、低3个剂量组对正常小鼠各脏器指数无显著性影响(p>0.05),单核-巨噬细胞功能结果为阴性;阳性对照组、低聚肽高剂量组能显著提高正常小鼠NK细胞活性(p<0.05),其结果为阳性。综上结果表明,远东拟沙丁鱼低聚肽对正常小鼠具有增强免疫力功能。
To evaluate the immune function of Sardinops sagax oligopeptide(SSO),the oligopeptide was prepared from Sardinops sagax by using enzymatic hydrolysis,centrifugation,filtration and spray drying,and its chemical composition was analyzed. Animal experiments were conducted to study immunological enhancement effect by the evaluation techniques of healthy food. The results showed that the molecular weight of SSO was mainly below 1000 u,accounting for 66. 4% of the total dry weight,and glutamic acid,lysine,aspartic acid and leucine were higher in the amino acid content. SSO was rich in selenium,which was 1. 66 mg/100 g. Compared with the negative control group,the proliferation rate of lymphocytes in normal mice was significantly higher in the positive control group and the SSO high-dose group(p < 0. 05),and the assay result of cellular immune function was positive. The spleen index and the number of plaque in normal mice could be improved significantly in the positive control group,the SSO high-dose group,the SSO medium-dose group and the SSO low-dose group(p < 0. 05),and the assay result of humoral immune function was positive. The positive control group,the SSO high-dose group,the SSO medium-dose group and the SSO low-dose group had no significant influence on the organ indexes of normal mice(p > 0. 05),and the assay result of mononuclear-macrophage function was negative. The positive control group and the SSO high-dose group could significantly increase the activity of NK cells in normal mice(p < 0. 05),and the assay result was positive. In conclusion,SSO can improve immune function in normal mice.
To evaluate the immune function of Sardinops sagax oligopeptide(SSO),the oligopeptide was prepared from Sardinops sagax by using enzymatic hydrolysis,centrifugation,filtration and spray drying,and its chemical composition was analyzed. Animal experiments were conducted to study immunological enhancement effect by the evaluation techniques of healthy food. The results showed that the molecular weight of SSO was mainly below 1000 u,accounting for 66. 4% of the total dry weight,and glutamic acid,lysine,aspartic acid and leucine were higher in the amino acid content. SSO was rich in selenium,which was 1. 66 mg/100 g. Compared with the negative control group,the proliferation rate of lymphocytes in normal mice was significantly higher in the positive control group and the SSO high-dose group(p < 0. 05),and the assay result of cellular immune function was positive. The spleen index and the number of plaque in normal mice could be improved significantly in the positive control group,the SSO high-dose group,the SSO medium-dose group and the SSO low-dose group(p < 0. 05),and the assay result of humoral immune function was positive. The positive control group,the SSO high-dose group,the SSO medium-dose group and the SSO low-dose group had no significant influence on the organ indexes of normal mice(p > 0. 05),and the assay result of mononuclear-macrophage function was negative. The positive control group and the SSO high-dose group could significantly increase the activity of NK cells in normal mice(p < 0. 05),and the assay result was positive. In conclusion,SSO can improve immune function in normal mice.
引文
[1]刘铭,刘玉环,王允圃,等.制备、纯化和鉴定生物活性肽的研究进展及应用[J].食品与发酵工业,2016,42(4):244-251.
[2]吕学泽,王雪敏,梁玉荣,等.免疫活性肽的研究进展[J].中国畜牧医,2011,38(4):170-172.
[3]程媛,曹慧,徐斐,等.食源性蛋白中免疫活性肽的研究进展[J].食品科学,2015,36(17):296-299.
[4]HANNU K.Milk-derived bioactive peptides:From science toapplications[J].Journal of Functional Foods,2009,1(2):177-187.
[5]MARTHA P,SAISLING A B,DARA O,et al.Potential bioactive effects of casein hydrolysates on human cultured cells[J].International Dairy Journal,2009,19(5):279-285.
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[7]KAYSER H,MEISEL H.Stimulation of human peripheral blood lymphocytes by bioactive peptides derived from bovine milk proteins[J].FEBS Letters,1996,383(1-2):18-20.
[8]代卉,乐国伟,孙进,等.小麦肽对受环磷酰胺免疫抑制小鼠的免疫调节及抗氧化功能[J].生物工程学报,2009,25(4):549-553.
[9]卢连华,周连奎,谢玮,等.大豆肽粉对低白细胞大鼠白细胞及NK细胞的影响[J].食品与药品,2014,16(4):262-265.
[10]于荣敏,严春艳,曲红艳,等.近年来海洋生物活性多肽的研究概况与展望[J].海洋通报,2004,23(3):87-88.
[11]王杏珠.日本远东拟沙丁鱼Sardinops melanostictus(Temniinek et Sehlegel)开发利用的新技术[J].现代水产渔业,1994,9(9):13-15.
[12]MATSUFUJI H,MATSUI T,SEKI E,et al.Angiotensin I-converting enzyme inhibitory peptides in an alkaline protease hydrolyzate derived from sardine muscle[J].Bioscience,Biotechnology and Biochemistry,1994,58(12):2 244-2 245.
[13]王晶晶,吉薇,吉宏武,等.远东拟沙丁鱼酶解产物的降血压效果评价及其ACE抑制肽的分离纯化与鉴定[J].食品与发酵工业,2016,42(4):63-68.
[14]GARCA-MORENO P,BATISTA I,PIRES C,et al.Antioxidant activity of protein hydrolysates obtained from discarded Mediterranean fish species[J].Food Research International,2014(65):469-476.
[15]李亚会,吉薇,吉宏武,等.远东拟沙丁鱼蛋白多肽(Sardinops sagax polypeptide)的制备及其对酪氨酸酶的抑制作用[J].食品工业科技,2016,37(3):58-62,68.
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[18]阮晓慧,韩军岐,张润光,等.食源性生物活性肽制备工艺、功能特性及应用研究进展[J].食品与发酵工业,2016,42(6):248-253.
[19]王晶,吴燕燕,杨贤庆.海洋贝类活性肽的研究现状和应用前景[J].食品工业科技,2013,34(13):346-349.
[20]QIAN B,XING M,GUI L,et al.Antioxidant,antihypertensive,and immunomodulatory activities of peptide fractions from fermented skim milk with Lactobacillus delbrueckii SSP bulgaricus LB3[J].Journal of Dairy Research,2011,78(1):72-79.
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[22]王晓芳,陈同辛.氨基酸与免疫功能的关系[J].国际儿科学杂志,2013,40(1):28-31.
[23]LI B,CHEN F,WANG X,et al.Isolation and identification of Antioxidative peptides from porcine collagen hydrolysate by Consecutive chromatography and electrospray ionization-mass spectrometry[J].Food Chemistry,2007,102(4):1 135-1 143.
[24]张英,董绍华.氨基酸清除活性氧自由基作用的研究[J].科技通报,1997,13(5):33-36.
[25]王真真,何慧,王驰,等.富硒玉米醒酒肽的制备及活性研究[J].食品工业科技,2013,34(18):239-242;248.
[26]李洁,王晓杰,邹素兰.胸腺肽的临床应用与研究进展[J].中国药房,2008,19(14):1 108-1 109.
[27]杨小军,左伟勇,陈伟华,等.面筋蛋白的胃蛋白酶酶解物对大鼠免疫功能的影响[J].南京农业大学学报,2004,27(4):69-72.
[28]张鸣镝,管晓,姚惠源.玉米胚芽蛋白酶解物对小鼠免疫功能的影响[J].食品科学,2007,28(2):302-305.
[29]李八方.功能食品与保健食品[M].青岛:中国海洋大学出版社,1997:11-27.
[30]杨贵贞.人体的免疫功能研究进展[J].新医学,1996,27(11):8-9.
[31]傅炜昕,董占双,李铁英,等.免疫活性地龙肽的制备及其对小鼠巨噬细胞活性的影响[J].微生物学杂志,2008,28(1):36-40.
[32]刘仁杰.林蛙油活性肽对小鼠免疫功能增强作用研究[D].长春:吉林农业大学,2016.
[2]吕学泽,王雪敏,梁玉荣,等.免疫活性肽的研究进展[J].中国畜牧医,2011,38(4):170-172.
[3]程媛,曹慧,徐斐,等.食源性蛋白中免疫活性肽的研究进展[J].食品科学,2015,36(17):296-299.
[4]HANNU K.Milk-derived bioactive peptides:From science toapplications[J].Journal of Functional Foods,2009,1(2):177-187.
[5]MARTHA P,SAISLING A B,DARA O,et al.Potential bioactive effects of casein hydrolysates on human cultured cells[J].International Dairy Journal,2009,19(5):279-285.
[6]JOLLES P,FIAT A M,MIGLOIRE-SAMOUR D,et al.Peptides from milk proteins implicated in antithrombosis and immunomoduation[C].New Perspectives in Infant Nutrition,Symposium Antwerp,1992:160-172.
[7]KAYSER H,MEISEL H.Stimulation of human peripheral blood lymphocytes by bioactive peptides derived from bovine milk proteins[J].FEBS Letters,1996,383(1-2):18-20.
[8]代卉,乐国伟,孙进,等.小麦肽对受环磷酰胺免疫抑制小鼠的免疫调节及抗氧化功能[J].生物工程学报,2009,25(4):549-553.
[9]卢连华,周连奎,谢玮,等.大豆肽粉对低白细胞大鼠白细胞及NK细胞的影响[J].食品与药品,2014,16(4):262-265.
[10]于荣敏,严春艳,曲红艳,等.近年来海洋生物活性多肽的研究概况与展望[J].海洋通报,2004,23(3):87-88.
[11]王杏珠.日本远东拟沙丁鱼Sardinops melanostictus(Temniinek et Sehlegel)开发利用的新技术[J].现代水产渔业,1994,9(9):13-15.
[12]MATSUFUJI H,MATSUI T,SEKI E,et al.Angiotensin I-converting enzyme inhibitory peptides in an alkaline protease hydrolyzate derived from sardine muscle[J].Bioscience,Biotechnology and Biochemistry,1994,58(12):2 244-2 245.
[13]王晶晶,吉薇,吉宏武,等.远东拟沙丁鱼酶解产物的降血压效果评价及其ACE抑制肽的分离纯化与鉴定[J].食品与发酵工业,2016,42(4):63-68.
[14]GARCA-MORENO P,BATISTA I,PIRES C,et al.Antioxidant activity of protein hydrolysates obtained from discarded Mediterranean fish species[J].Food Research International,2014(65):469-476.
[15]李亚会,吉薇,吉宏武,等.远东拟沙丁鱼蛋白多肽(Sardinops sagax polypeptide)的制备及其对酪氨酸酶的抑制作用[J].食品工业科技,2016,37(3):58-62,68.
[16]侯虎.鳕鱼免疫活性肽的可控制备及其免疫活性研究[D].青岛:中国海洋大学,2011.
[17]林端权,郭泽镔,张怡,等.海洋生物活性肽的研究进展[J].食品工业科技,2016,37(18):367-373.
[18]阮晓慧,韩军岐,张润光,等.食源性生物活性肽制备工艺、功能特性及应用研究进展[J].食品与发酵工业,2016,42(6):248-253.
[19]王晶,吴燕燕,杨贤庆.海洋贝类活性肽的研究现状和应用前景[J].食品工业科技,2013,34(13):346-349.
[20]QIAN B,XING M,GUI L,et al.Antioxidant,antihypertensive,and immunomodulatory activities of peptide fractions from fermented skim milk with Lactobacillus delbrueckii SSP bulgaricus LB3[J].Journal of Dairy Research,2011,78(1):72-79.
[21]STUKNYTE M,DE N I,GUGLIELMETTI S,et al.Potential immunomodulatory activity of bovine casein hydrolysates produced after digestion with proteinases of lactic acid bacteria[J].International Dairy Journal,2011,21(10):763-769.
[22]王晓芳,陈同辛.氨基酸与免疫功能的关系[J].国际儿科学杂志,2013,40(1):28-31.
[23]LI B,CHEN F,WANG X,et al.Isolation and identification of Antioxidative peptides from porcine collagen hydrolysate by Consecutive chromatography and electrospray ionization-mass spectrometry[J].Food Chemistry,2007,102(4):1 135-1 143.
[24]张英,董绍华.氨基酸清除活性氧自由基作用的研究[J].科技通报,1997,13(5):33-36.
[25]王真真,何慧,王驰,等.富硒玉米醒酒肽的制备及活性研究[J].食品工业科技,2013,34(18):239-242;248.
[26]李洁,王晓杰,邹素兰.胸腺肽的临床应用与研究进展[J].中国药房,2008,19(14):1 108-1 109.
[27]杨小军,左伟勇,陈伟华,等.面筋蛋白的胃蛋白酶酶解物对大鼠免疫功能的影响[J].南京农业大学学报,2004,27(4):69-72.
[28]张鸣镝,管晓,姚惠源.玉米胚芽蛋白酶解物对小鼠免疫功能的影响[J].食品科学,2007,28(2):302-305.
[29]李八方.功能食品与保健食品[M].青岛:中国海洋大学出版社,1997:11-27.
[30]杨贵贞.人体的免疫功能研究进展[J].新医学,1996,27(11):8-9.
[31]傅炜昕,董占双,李铁英,等.免疫活性地龙肽的制备及其对小鼠巨噬细胞活性的影响[J].微生物学杂志,2008,28(1):36-40.
[32]刘仁杰.林蛙油活性肽对小鼠免疫功能增强作用研究[D].长春:吉林农业大学,2016.