CN114106092B - 一种具有ace抑制作用的活性多肽及其应用 - Google Patents
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Abstract
本发明公开了一种具有ACE抑制作用的活性多肽，其氨基酸序列为WGAP。本发明一种具有ACE抑制作用的活性多肽对ACE的活性具有持续稳定的抑制作用，并具有良好的降血压效果，因此可作为活性成分用于食品、保健品以及降血压药物中，具有良好的潜力和应用前景。
Description
技术领域
本发明属于生物医药技术领域，具体的说，涉及一种具有ACE抑制作用的活性多肽及其应用。
背景技术
高血压是最常见的心血管疾病之一，也是导致脑卒中、冠心病、心力衰竭等疾病的重要危险因素。根据2020年世界卫生组织的统计，全世界约有11.3亿人患有高血压，已经严重威胁人类的健康。血管紧张素转换酶（Angiotensin-I converting enzyme, ACE）是一种二肽羧基金属蛋白酶，在肾素-血管紧张素***（Renin-Angiotensin System, RAS）和激肽释放酶-激肽***（Kallikrein-Kinin System, KKS）中起着重要的调控作用。ACE抑制剂可通过抑制ACE活性，阻止血管紧张素I转化为血管紧张素II，维持血管紧张素缓激肽的活性来降低血压。目前，常用于治疗高血压的ACE抑制剂有卡托普利、依那普利、福辛普利、拉米普利等。虽效果明显，但长期服用副作用较大，包括咳嗽、味觉变形、皮疹、肾脏功能下降以及患肺癌风险增加等。因此，研究重点已转向寻找对健康有益且无副作用的天然ACE抑制剂。
食品源活性肽具有易溶解性、无致敏性和良好的ADMET（Absorption, Distribution, Metabolism, Excretion and Toxicity）特性，因而越来越受到人们的关注。近年来，具有多种功能的食品源生物肽被发现，例如ACE抑制肽，抗炎肽，二肽基肽酶IV抑制肽和抗氧化肽等。目前，国内外学者已发现大量动植物中含有抑制ACE的生物活性肽，尤其是在各种常食用动物当中，如鸡、鸭、鱼、猪、牛等。
新西兰白兔作为我国饲养最多的食用兔类，是一种高营养价值的功能性肉类食品。兔肉富含蛋白质、矿物质、维生素、卵磷脂和多不饱和脂肪酸，可以为消费者提供生物活性物质，经常食用可以对控制心血管疾病和其他慢性疾病起到关键作用。目前，国内外对兔肉的研究主要聚焦于营养价值，尚无有关兔肉源ACE抑制肽及其作用机制的研究报道。
发明内容
为解决以上技术问题，本发明的目的在于提供一种来源于兔肉、对 ACE的活性具有持续稳定的抑制作用、具有良好降血压效果的 ACE 抑制肽。
一、一种具有ACE抑制作用的活性多肽，其关键在于：其氨基酸序列为Trp-Gly-Ala-Pro（WGAP）。
二、一种上述具有ACE抑制作用的活性多肽在制备ACE抑制药物中的应用。
三、一种上述的具有ACE抑制作用的活性多肽在制备降血压药物中的应用。
四、一种具有ACE抑制作用的药物，其关键在于：其活性成分含有上述的具有ACE抑制作用的活性多肽。
五、一种具有降血压作用的药物，其关键在于：其活性成分含有上述的具有ACE抑制作用的活性多肽。
有益效果：
本发明一种具有ACE抑制作用的活性多肽，对ACE的活性具有持续稳定的抑制作用，并具有良好的降血压效果，因此可作为活性成分用于食品、保健品以及降血压药物中，具有良好的潜力和应用前景。
附图说明
图1为馏分F-4的反相高效液相色谱图；
图2为不同浓度的活性多肽在体外胃肠消化前后的ACE抑制活性；
图3为单次口服生理盐水、卡托普利（20 mg/kg）和WGAP（25、50和100 mg/kg）对高血压大鼠收缩压影响的线型图（数据为平均值±标准差（n = 6）；×表示P<0.05卡托普利组与生理盐水组，××表示P<0.01卡托普利组与生理盐水组，*表示P<0.05 25 mg/kg WGAP组与生理盐水组，**表示P<0.01 25 mg/kg WGAP组与生理盐水组，#表示P<0.05 50 mg/kgWGAP组与生理盐水组，##表示P<0.01 50 mg/kg WGAP组与生理盐水组，△表示P<0.05 100mg/kg WGAP组与生理盐水组，△△表示P<0.01 100 mg/kg WGAP组与生理盐水组）。
具体
下面结合实施例和附图对本发明作进一步说明。
实施例
将兔肉剁碎，用蒸馏水将浓度配至50mg/mL，并使用组织匀浆器匀浆2 min。匀浆液在沸水浴中灭活10 min。冷却后，使用五种不同的酶（碱性蛋白酶：50℃，pH7.0；胃蛋白酶：37℃，pH2.0；胰蛋白酶：37℃，pH8.0；复合蛋白酶：50℃，pH7.0；菠萝蛋白酶：55℃，pH7.0）在最佳条件下水解2、3、4、5或6小时，酶与兔肉的质量比为1:10。在水解过程中，反应混合物的pH值通过加入0.5N NaOH或0.5N HCl来保持。然后将混合物煮沸10分钟以终止反应，迅速冷却到室温，并调整到pH7.0。然后用TGL-20离心机在4℃条件下以10,000 r/min离心20 min。用滤纸过滤上清液，在冻干机中冻干，得到兔肉蛋白水解物（RMPH）。测量每个RMPH的ACE抑制活性，将活性最高的RMPH储存在-20℃以备进一步研究。随后采用超滤法、Sephadex G-15凝胶过滤层析（GFC）、高效液相色谱（RP-HPLC）对酶解物中功效组分进行筛选，最终找出F4-4具有最强ACE抑制活性（最终结果如图1）；然后采用液相色谱-串联质谱（HPLC-MS/MS）对功效肽组分进行鉴定，确定多肽氨基酸序列为Trp-Gly-Ala-Pro，即WGAP。
二、ACE抑制活性检测测定WGAP对ACE活性的抑制作用。在37℃下，将20µL的WGAP溶液与ACE溶液（20µL，25mU/mL）在50mM硼酸盐钠缓冲液（含0.3M NaCl，pH8.3）中预孵化10分钟。然后将马脲酰组氨酰亮氨酸 (HHL)溶液（20 µL，5 mM）加入到反应混合物中，并在37℃下孵育30min。加入100 µL的1 M HCl终止反应。再加入乙酸乙酯 (1 mL)，摇动后，并在10,000 r/min和4°C条件下离心10 min。取上层，在60℃下真空蒸发至干。将残余物溶于100 µL一级水，在228 nm处测量吸光度。ACE的抑制活性根据公式计算：
ACE 抑制活性(%) = (A对照– A样品)/(A对照– A空白)
其中A对照是没有抑制剂时的吸光度，A样品是反应混合物的吸光度，A空白是缓冲液的吸光度。IC50值被定义为对ACE活性造成50%抑制的抑制剂的浓度。
结果显示，多肽WGAP表现出的ACE抑制IC50为140.70±4.51 μM。
三、体外的消化稳定性将WGAP溶解在0.1 M KCl中，并用1 M HCl调节到pH值2.0。加入胃蛋白酶（E/S 2%，w/w），在37 ℃水浴中消化多肽2 h。通过在100℃下加热10 min停止反应，用2 M NaOH快速调节pH值至8.0。然后加入胰蛋白酶（E/S 2%，w/w），在37℃孵育3小时。再次在100℃下灭活10分钟。冷却到室温后，将水解液调整到pH7.0，并以10,000 r/min离心10分钟。最后，对上清液的ACE抑制活性进行检测，并与消化前的肽进行比较。结果如图2所示，用胃蛋白酶和胰蛋白酶处理后，WGAP的ACE抑制活性没有明显变化（p>0.05）。这些结果表明，WGAP对胃肠道消化有抵抗力，可在体内表现出降血压的作用。
四、体内的抗高血压作用雄性SD大鼠（10周，体重250±20 g）由中国重庆医科大学实验动物中心提供，在标准条件下（12小时光照/黑暗周期，22±1℃）饲养，自由采食和饮水。调理一周后，大鼠腹腔注射15mg/kg L-N-硝基精氨酸，每天一次，持续四周，以在实验开始前建立高血压模型。随后，将大鼠随机分为5个治疗组，每组6只：阳性对照组、阴性对照组、高、中、低剂量组。阳性对照组的大鼠口服卡托普利（20 mg/kg），阴性对照组的大鼠口服生理盐水。高、中、低剂量组的大鼠分别以100、50和25 mg/kg体重的剂量口服WGAP。使用无创血压测量分析***在给药后0、2、4、6、8、10和12小时测量收缩压（SBP）。结果如图3所示，阴性对照组（0.9%生理盐水）在给药后12小时内没有表现出SBP的下降，而阳性对照组（卡托普利，20 mg/kg）则表现出明显的下降（P< 0.01）。卡托普利给药6小时后，SBP下降了41.79±2.34 mmHg（P＜0.01）。然而，WGAP处理组（25、50和100 mg/kg）仅在4小时后就表现出SBP的下降（P＜0.01），分别为23.45±1.63、32.85±1.94和42.66±2.87 mmHg（图3），表明WGAP的吸收比卡托普利好。此外，SBP的变化程度是呈WGAP剂量依赖性的。12 h后，大鼠的SBP在高剂量WGAP组中明显降低了13.80±1.24mmHg（P＜0.01），在卡托普利组中降低了9.28±0.76 mmHg（P＜0.05），但与阴性对照组相比，中剂量和低剂量WGAP组没有明显差异（P＞0.05）。
因此，WGAP在高血压大鼠口服后能显著降低SBP，是一种潜在的天然ACE抑制肽。
最后需要说明的是，上述描述仅仅为本发明的优选实施例，本领域的普通技术人员在本发明的启示下，在不违背本发明宗旨及权利要求的前提下，可以做出多种类似的表示，这样的变换均落入本发明的保护范围之内。
Claims (5)
1.一种具有ACE抑制作用的活性多肽，其特征在于：其氨基酸序列为WGAP。
2.一种权利要求1所述的具有ACE抑制作用的活性多肽在制备ACE抑制药物中的应用。
3.一种权利要求1所述的具有ACE抑制作用的活性多肽在制备降血压药物中的应用。
4.一种具有ACE抑制作用的药物，其特征在于：其活性成分含有权利要求1所述的具有ACE抑制作用的活性多肽。
5.一种具有降血压作用的药物，其特征在于：其活性成分含有权利要求1所述的具有ACE抑制作用的活性多肽。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004067519A (ja) * | 2002-08-01 | 2004-03-04 | Nippon Meat Packers Inc | アンジオテンシン変換酵素阻害ペプチド |
| CN101570568A (zh) * | 2009-06-15 | 2009-11-04 | 东北农业大学 | 一种发酵乳中的ace抑制肽及其制备方法 |
| CN104945502A (zh) * | 2015-06-30 | 2015-09-30 | 石狮海星食品有限公司 | Ace抑制五肽 |
| CN108129561A (zh) * | 2017-12-06 | 2018-06-08 | 渤海大学 | 一种ace抑制肽 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004067519A (ja) * | 2002-08-01 | 2004-03-04 | Nippon Meat Packers Inc | アンジオテンシン変換酵素阻害ペプチド |
| CN101570568A (zh) * | 2009-06-15 | 2009-11-04 | 东北农业大学 | 一种发酵乳中的ace抑制肽及其制备方法 |
| CN104945502A (zh) * | 2015-06-30 | 2015-09-30 | 石狮海星食品有限公司 | Ace抑制五肽 |
| CN108129561A (zh) * | 2017-12-06 | 2018-06-08 | 渤海大学 | 一种ace抑制肽 |
Non-Patent Citations (1)
| Title |
|---|
| Characterization and antioxidant activity determination of neutral and acidic polysaccharides from panax ginseng C.A. meyer;Hyung Min Kim et al;《Molecules》;第25卷(第4期);全文 * |
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