大豆低聚肽对自发性高血压大鼠血压及血浆血管紧张素的影响
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摘要
目的:探究大豆低聚肽对于大鼠血压及血浆血管紧张素的影响。方法:采用酶法制备大豆低聚肽,随后进行体外血管紧张素Ⅰ转换酶(angiotensin Ⅰ converting enzyme,ACE)活性抑制实验及体内实验,实验设正常对照组(正常血压大鼠(WKY大鼠)饲喂高剂量大豆低聚肽)、阴性对照组(自发性高血压大鼠(SHR大鼠))、阳性对照组(SHR大鼠饲喂卡托普利)和低、中、高剂量组(SHR大鼠饲喂0.90、1.80、4.50 g/kg大豆低聚肽),喂养30d后,观察大鼠血压、心率、尿蛋白质量、血管紧张素Ⅱ质量浓度等指标的变化。结果:大豆蛋白最佳酶解条件为:碱性蛋白酶和中性蛋白酶质量分数各0.1%、50℃酶解4 h,此条件下10 mg/mL大豆低聚肽ACE活性抑制率达到71.2%。经饲喂不同剂量大豆低聚肽30 d后,WKY大鼠血压变化不显著(P>0.05),SHR大鼠血压有下降趋势;饲喂高剂量大豆低聚肽在第4周时可以显著降低SHR大鼠血压和血管紧张素Ⅱ质量浓度(P<0.05),但对于大鼠心率和尿蛋白质量无显著影响(P>0.05)。结论:体外实验证明,与大豆蛋白相比,大豆低聚肽对于ACE活性的抑制效果更好;体内实验证明大豆低聚肽可以降低SHR大鼠的血压和血管紧张素Ⅱ质量浓度,且对正常大鼠血压无明显影响,推测大豆低聚肽通过抑制ACE活性的方式发挥降压功效。
Purpose: To investigate the effect of soybean oligopeptides on blood pressure and plasma angiotensin levels in rats. Methods: Soybean oligopeptides were prepared by enzymatic treatment, and its angiotensin Ⅰ converting enzyme(ACE) inhibitory activity was evaluated both in vitro and in vivo experiments. Rats were divided into six groups: normal control(Wistar-Kyoto(WKY) rats with normal blood pressure rats fed high-dose soybean oligopeptides), negative control(spontaneously hypertensive(SHR) rats), positive control(SHR rats fed captopril), low, medium and high-dose groups(SHR rats fed 0.90, 1.80, 4.50 g/kg soybean oligopeptides). The feeding experiment lasted for 30 days. The changes in blood pressure, heart rate, urinary protein content and plasma angiotensin Ⅱ concentration were determined during the experimental period. Results: The optimal conditions for enzymatic hydrolysis of soybean protein were found to be hydrolysis at 50 ℃ for 4 h with a mixture of alkaline protease and neutral protease at a dosage of 0.1% each. The percentage inhibition of ACE activity by soybean oligopeptides at 10 mg/mL prepared using the optimized conditions was up to 71.2%. After 30 days of feeding, there was no significant difference in blood pressure of WKY rats(P > 0.05), while the blood pressure of SHR rats had a decreasing trend in all dose groups of soybean oligopeptides. High-dose soybean oligopeptides significantly reduced blood pressure and mass concentration of angiotensin Ⅱ in SHR rats at the end of the fourth week(P < 0.05), but had no significant effect on rat heart rate or urinary protein content(P > 0.05). Conclusion: Compared with soybean protein,soybean oligopeptides had better inhibitory effect on ACE activity, and could reduce blood pressure and plasma angiotensin II levels in SHR rats but without significant effect on blood pressure in normal rats. Therefore, soybean oligopeptides can exert antihypertensive effect perhaps by inhibiting ACE activity.
Purpose: To investigate the effect of soybean oligopeptides on blood pressure and plasma angiotensin levels in rats. Methods: Soybean oligopeptides were prepared by enzymatic treatment, and its angiotensin Ⅰ converting enzyme(ACE) inhibitory activity was evaluated both in vitro and in vivo experiments. Rats were divided into six groups: normal control(Wistar-Kyoto(WKY) rats with normal blood pressure rats fed high-dose soybean oligopeptides), negative control(spontaneously hypertensive(SHR) rats), positive control(SHR rats fed captopril), low, medium and high-dose groups(SHR rats fed 0.90, 1.80, 4.50 g/kg soybean oligopeptides). The feeding experiment lasted for 30 days. The changes in blood pressure, heart rate, urinary protein content and plasma angiotensin Ⅱ concentration were determined during the experimental period. Results: The optimal conditions for enzymatic hydrolysis of soybean protein were found to be hydrolysis at 50 ℃ for 4 h with a mixture of alkaline protease and neutral protease at a dosage of 0.1% each. The percentage inhibition of ACE activity by soybean oligopeptides at 10 mg/mL prepared using the optimized conditions was up to 71.2%. After 30 days of feeding, there was no significant difference in blood pressure of WKY rats(P > 0.05), while the blood pressure of SHR rats had a decreasing trend in all dose groups of soybean oligopeptides. High-dose soybean oligopeptides significantly reduced blood pressure and mass concentration of angiotensin Ⅱ in SHR rats at the end of the fourth week(P < 0.05), but had no significant effect on rat heart rate or urinary protein content(P > 0.05). Conclusion: Compared with soybean protein,soybean oligopeptides had better inhibitory effect on ACE activity, and could reduce blood pressure and plasma angiotensin II levels in SHR rats but without significant effect on blood pressure in normal rats. Therefore, soybean oligopeptides can exert antihypertensive effect perhaps by inhibiting ACE activity.
引文
[1]韩飞,于婷婷,周孟良.酶法生产大豆蛋白A C E抑制肽的研究[J].食品科学,2008,29(11):369-374.DOI:10.3321/j.issn:1002-6630.2008.11.084.
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[3]吕育新,梁金钟.大豆低聚肽的研究进展[J].大豆科技,2013(2):37-39.DOI:10.3969/j.issn.1674-3547.2013.02.013.
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[5]郭玉华,李钰金,吴新颖.大豆多肽的应用进展及前景[J].粮食加工,2010,35(5):60-63.DOI:10.3969/j.issn.1007-6395.2010.05.018.
[6]王立博,陈复生.大豆活性肽生理保健功能研究进展[J].食品与机械,2016,32(2):198-201.DOI:10.13652/j.issn.1003-5788.2016.02.047.
[7]张连慧,贺寅,刘新旗.大豆肽制备研究进展及其在食品中的应用[J].食品工业科技,2012,33(24):423-425;429.DOI:10.13386/j.issn1002-0306.2012.24.063.
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[9]HANAFI M A,HASHIM S N,CHAY S Y,et al.High angiotensin-Iconverting enzyme(ACE)inhibitory activity of alcalase-digested green soybean hydrolysates[J].Food Research International,2018,106:589-597.DOI:10.1016/j.foodres.2018.01.030.
[10]JIMSHEENA V K,GOWDA L R.Angiotensin I-converting enzyme(ACE)inhibitory peptides derived from arachin by simulated gastric digestion[J].Food Chemistry,2011,125(2):561-569.DOI:10.1016/j.foodchem.2010.09.048.
[11]杨湘华,李江.食源性降血压肽构效关系与制备研究进展[J].食品研究与开发,2016,37(3):217-220.DOI:10.3969/j.issn.1005-6521.2016.03.054.
[12]刘新旗,涂丛慧,张连慧,等.大豆蛋白的营养保健功能研究现状[J].北京工商大学学报(自然科学版),2012,30(2):1-6.DOI:10.3969/j.issn.1671-1513.2012.02.001.
[13]崔和平,郭兴凤.食源性降血压肽制备研究进展[J].粮食与油脂,2013,26(2):5-8.DOI:10.3969/j.issn.1008-9578.2013.02.004.
[14]MAMILLA R K,MISHRA V K.Effect of germination on antioxidant and ACE inhibitory activities of legumes[J].LWT-Food Science and Technology,2017,75:51-58.DOI:10.1016/j.lwt.2016.08.036.
[15]SANJUKTA S,RAI A K.Production of bioactive peptides during soybean fermentation and their potential health benefits[J].Trends in Food Science&Technology,2016,50:1-10.DOI:10.1016/j.tifs.2016.01.010.
[16]KUBA M,TANA C,TAWATA S,et al.Production of angiotensin I-converting enzyme inhibitory peptides from soybean protein with Monascus purpureus acid proteinase[J].Process Biochemistry,2005,40(6):2191-2196.DOI:10.1016/j.procbio.2004.08.010.
[17]ALUKO R E.Structure and function of plant protein-derived antihypertensive peptides[J].Current Opinion in Food Science,2015,4:44-50.DOI:10.1016/j.cofs.2015.05.002.
[18]范远景,姬莹莹,张焱.大豆蛋白酶解肽的分子量分布及抑制ACE活性关系研究[J].食品科学,2007,28(10):57-61.DOI:10.3321/j.issn:1002-6630.2007.10.008.
[19]王升光,于帅,孟凡刚,等.酶法制备大豆肽的相对分子量分布及降压作用研究[J].食品工业科技,2018,39(1):46-51.DOI:10.13386/j.issn.1002-0306.2018.01.009.
[20]BOYE J I,ROUFIK S,PESTA N,et al.Angiotensin I-converting enzyme inhibitory properties and SDS-PAGE of red lentil protein hydrolysates[J].LWT-Food Science and Technology,2010,43(6):987-991.DOI:10.1016/j.lwt.2010.01.014.
[21]BOSCHIN G,SCIGLIUOLO G M,RESTA D,et al.ACE-inhibitory activity of enzymatic protein hydrolysates from lupin and other legumes[J].Food Chemistry,2014,145(7):34-40.DOI:10.1016/j.foodchem.2013.07.076.
[22]WU Jianping,ALUKO R E,MUIR A D.Improved method for direct high-performance liquid chromatography assay of angiotensinconverting enzyme-catalyzed reactions[J].Journal of Chromatography A,2002,950(1/2):125-130.DOI:10.1016/S0021-9673(02)00052-3.
[23]KIVIM?KI A S,SILTARI A,EHLERS P I,et al.Lingonberry juice lowers blood pressure of spontaneously hypertensive rats(SHR)[J].Journal of Functional Foods,2013,5(3):1432-1440.DOI:10.1016/j.jff.2013.05.012.
[24]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,2010:1535.
[25]李晓红,宋福印.丹参多糖对SHR高血压模型大鼠血压和COX-2基因表达的影响[J].北京中医药,2017,36(8):706-709.DOI:10.16025/j.1674-1307.2017.08.012.
[26]李超民,罗薇,李慧,等.人参皂苷Rg2对SHR大鼠血压及肾脏MMP2、MMP9表达的影响[J].心脏杂志,2017,29(1):44-47.DOI:10.13191/j.chj.2017.0012.
[27]邓九零,陈亮,寿旗扬,等.益气活血方对糖尿病GK大鼠肾脏保护作用的研究[J].中国中药杂志,2018,43(11):2372-2377.DOI:10.19540/j.cnki.cjcmm.20180327.004.
[28]DE ALMEIDA C L B,CECHINEL-FILHO V,BOEING T,et al.Prolonged diuretic and saluretic effect of nothofagin isolated from Leandra dasytricha(A.Gray)Cogn.leaves in normotensive and hypertensive rats:role of antioxidant system and renal protection[J].Chemico-Biological Interactions,2018,279:227-233.DOI:10.1016/j.cbi.2017.11.021.
[29]CAIN M D,COGHLAN J P,CATT K J.Measurement of angiotensin II in blood by radioimmunoassay[J].Clinica Chimica Acta,1972,39(1):21-34.DOI:10.1016/0009-8981(72)90295-1.
[30]杨雯晴,蒋海强,李运伦,等.卡托普利干预自发性高血压大鼠血清代谢组学研究[J].中国药理学通报,2016,32(7):998-1003.DOI:10.3969/j.issn.1001-1978.2016.07.022.
[31]WU Jianping,DING Xiaolin.Hypotensive and physiological effect of angiotensin converting enzyme inhibitory peptides derived from soy protein on spontaneously hypertensive rats[J].Journal of Agricultural and Food Chemistry,2001,49(1):501-506.DOI:10.1021/jf000695n.
[32]卢静,赵春燕,张学新,等.大豆肽对大鼠血压及血浆血管紧张素的影响[J].营养学报,2008,30(5):508-511.DOI:10.13325/j.cnki.acta.nutr.sin.2008.05.020.
[33]孙宁玲,霍勇,黄峻.中国高血压患者心率现状调查[J].中华高血压杂志,2015,23(10):934-939.DOI:10.16439/j.cnki.1673-7245.2015.10.013.
[34]JAHANDIDEH F,CHAKRABARTI S,MAJUMDER K,et al.Egg white protein hydrolysate reduces blood pressure,improves vascular relaxation and modifies aortic angiotensin II receptors expression in spontaneously hypertensive rats[J].Journal of Functional Foods,2016,27:667-673.DOI:10.1016/j.jff.2016.10.019.
[35]于海荣,曲正阳,王一帆,等.决明子蒽醌苷对高血压大鼠血压、尿白蛋白及β2-微球蛋白的影响[J].中国老年学杂志,2015,35(21):6041-6042.DOI:10.3969/j.issn.1005-9202.2015.21.016.
[36]王晓玲,张先杰,黄霜枝,等.葡萄籽原花青素对自发性高血压大鼠肾损伤的保护作用[J].中药药理与临床,2017,33(5):43-47.DOI:10.13412/j.cnki.zyyl.2017.05.012.
[37]孟月,饶欢,陶莎,等.豆源降血压肽制备及活性评价方法的研究进展[J].粮食与油脂,2016,29(7):5-9.DOI:10.3969/j.issn.1008-9578.2016.07.002.
[2]EBERT A W,CHANG C H,YAN M R,et al.Nutritional composition of mungbean and soybean sprouts compared to their adult growth stage[J].Food Chemistry,2017,237:15-22.DOI:10.3321/j.issn:1002-6630.2008.11.084.
[3]吕育新,梁金钟.大豆低聚肽的研究进展[J].大豆科技,2013(2):37-39.DOI:10.3969/j.issn.1674-3547.2013.02.013.
[4]国家质量监督检验检疫总局.大豆肽粉:GB/T 22492-2008[S].北京:中国标准出版社,2009:1-3.
[5]郭玉华,李钰金,吴新颖.大豆多肽的应用进展及前景[J].粮食加工,2010,35(5):60-63.DOI:10.3969/j.issn.1007-6395.2010.05.018.
[6]王立博,陈复生.大豆活性肽生理保健功能研究进展[J].食品与机械,2016,32(2):198-201.DOI:10.13652/j.issn.1003-5788.2016.02.047.
[7]张连慧,贺寅,刘新旗.大豆肽制备研究进展及其在食品中的应用[J].食品工业科技,2012,33(24):423-425;429.DOI:10.13386/j.issn1002-0306.2012.24.063.
[8]张连慧,贺寅,刘新旗.大豆肽的研究进展及其发展前景[J].食品工业科技,2012,33(22):406-408;413.DOI:10.13386/j.issn1002-0306.2012.22.093.
[9]HANAFI M A,HASHIM S N,CHAY S Y,et al.High angiotensin-Iconverting enzyme(ACE)inhibitory activity of alcalase-digested green soybean hydrolysates[J].Food Research International,2018,106:589-597.DOI:10.1016/j.foodres.2018.01.030.
[10]JIMSHEENA V K,GOWDA L R.Angiotensin I-converting enzyme(ACE)inhibitory peptides derived from arachin by simulated gastric digestion[J].Food Chemistry,2011,125(2):561-569.DOI:10.1016/j.foodchem.2010.09.048.
[11]杨湘华,李江.食源性降血压肽构效关系与制备研究进展[J].食品研究与开发,2016,37(3):217-220.DOI:10.3969/j.issn.1005-6521.2016.03.054.
[12]刘新旗,涂丛慧,张连慧,等.大豆蛋白的营养保健功能研究现状[J].北京工商大学学报(自然科学版),2012,30(2):1-6.DOI:10.3969/j.issn.1671-1513.2012.02.001.
[13]崔和平,郭兴凤.食源性降血压肽制备研究进展[J].粮食与油脂,2013,26(2):5-8.DOI:10.3969/j.issn.1008-9578.2013.02.004.
[14]MAMILLA R K,MISHRA V K.Effect of germination on antioxidant and ACE inhibitory activities of legumes[J].LWT-Food Science and Technology,2017,75:51-58.DOI:10.1016/j.lwt.2016.08.036.
[15]SANJUKTA S,RAI A K.Production of bioactive peptides during soybean fermentation and their potential health benefits[J].Trends in Food Science&Technology,2016,50:1-10.DOI:10.1016/j.tifs.2016.01.010.
[16]KUBA M,TANA C,TAWATA S,et al.Production of angiotensin I-converting enzyme inhibitory peptides from soybean protein with Monascus purpureus acid proteinase[J].Process Biochemistry,2005,40(6):2191-2196.DOI:10.1016/j.procbio.2004.08.010.
[17]ALUKO R E.Structure and function of plant protein-derived antihypertensive peptides[J].Current Opinion in Food Science,2015,4:44-50.DOI:10.1016/j.cofs.2015.05.002.
[18]范远景,姬莹莹,张焱.大豆蛋白酶解肽的分子量分布及抑制ACE活性关系研究[J].食品科学,2007,28(10):57-61.DOI:10.3321/j.issn:1002-6630.2007.10.008.
[19]王升光,于帅,孟凡刚,等.酶法制备大豆肽的相对分子量分布及降压作用研究[J].食品工业科技,2018,39(1):46-51.DOI:10.13386/j.issn.1002-0306.2018.01.009.
[20]BOYE J I,ROUFIK S,PESTA N,et al.Angiotensin I-converting enzyme inhibitory properties and SDS-PAGE of red lentil protein hydrolysates[J].LWT-Food Science and Technology,2010,43(6):987-991.DOI:10.1016/j.lwt.2010.01.014.
[21]BOSCHIN G,SCIGLIUOLO G M,RESTA D,et al.ACE-inhibitory activity of enzymatic protein hydrolysates from lupin and other legumes[J].Food Chemistry,2014,145(7):34-40.DOI:10.1016/j.foodchem.2013.07.076.
[22]WU Jianping,ALUKO R E,MUIR A D.Improved method for direct high-performance liquid chromatography assay of angiotensinconverting enzyme-catalyzed reactions[J].Journal of Chromatography A,2002,950(1/2):125-130.DOI:10.1016/S0021-9673(02)00052-3.
[23]KIVIM?KI A S,SILTARI A,EHLERS P I,et al.Lingonberry juice lowers blood pressure of spontaneously hypertensive rats(SHR)[J].Journal of Functional Foods,2013,5(3):1432-1440.DOI:10.1016/j.jff.2013.05.012.
[24]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,2010:1535.
[25]李晓红,宋福印.丹参多糖对SHR高血压模型大鼠血压和COX-2基因表达的影响[J].北京中医药,2017,36(8):706-709.DOI:10.16025/j.1674-1307.2017.08.012.
[26]李超民,罗薇,李慧,等.人参皂苷Rg2对SHR大鼠血压及肾脏MMP2、MMP9表达的影响[J].心脏杂志,2017,29(1):44-47.DOI:10.13191/j.chj.2017.0012.
[27]邓九零,陈亮,寿旗扬,等.益气活血方对糖尿病GK大鼠肾脏保护作用的研究[J].中国中药杂志,2018,43(11):2372-2377.DOI:10.19540/j.cnki.cjcmm.20180327.004.
[28]DE ALMEIDA C L B,CECHINEL-FILHO V,BOEING T,et al.Prolonged diuretic and saluretic effect of nothofagin isolated from Leandra dasytricha(A.Gray)Cogn.leaves in normotensive and hypertensive rats:role of antioxidant system and renal protection[J].Chemico-Biological Interactions,2018,279:227-233.DOI:10.1016/j.cbi.2017.11.021.
[29]CAIN M D,COGHLAN J P,CATT K J.Measurement of angiotensin II in blood by radioimmunoassay[J].Clinica Chimica Acta,1972,39(1):21-34.DOI:10.1016/0009-8981(72)90295-1.
[30]杨雯晴,蒋海强,李运伦,等.卡托普利干预自发性高血压大鼠血清代谢组学研究[J].中国药理学通报,2016,32(7):998-1003.DOI:10.3969/j.issn.1001-1978.2016.07.022.
[31]WU Jianping,DING Xiaolin.Hypotensive and physiological effect of angiotensin converting enzyme inhibitory peptides derived from soy protein on spontaneously hypertensive rats[J].Journal of Agricultural and Food Chemistry,2001,49(1):501-506.DOI:10.1021/jf000695n.
[32]卢静,赵春燕,张学新,等.大豆肽对大鼠血压及血浆血管紧张素的影响[J].营养学报,2008,30(5):508-511.DOI:10.13325/j.cnki.acta.nutr.sin.2008.05.020.
[33]孙宁玲,霍勇,黄峻.中国高血压患者心率现状调查[J].中华高血压杂志,2015,23(10):934-939.DOI:10.16439/j.cnki.1673-7245.2015.10.013.
[34]JAHANDIDEH F,CHAKRABARTI S,MAJUMDER K,et al.Egg white protein hydrolysate reduces blood pressure,improves vascular relaxation and modifies aortic angiotensin II receptors expression in spontaneously hypertensive rats[J].Journal of Functional Foods,2016,27:667-673.DOI:10.1016/j.jff.2016.10.019.
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