CN116514904A - 一种缓解结肠炎的活性肽vpp及其应用 - Google Patents
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Abstract
本发明公开了一种缓解结肠炎的活性肽VPP及其应用。本发明所提供的活性肽VPP对动物无毒副作用。采用本发明所提供的VPP能够显著降低DSS诱导结肠炎期间小鼠的DAI指数，减轻结肠的缩短。采用本发明所提供的VPP，能够缓解DSS诱导的小鼠结肠组织会出现的明显损伤，包括个别肠腺消失被增生的结缔组织取代，杯状细胞数量减少，周围伴有少量的淋巴细胞浸润。与模型组小鼠相比，VPP干预组小鼠的结肠组织中促炎细胞因子TNF‑α、IL‑6、IL‑8和IL‑1β的浓度显著降低。VPP干预使血清免疫球蛋白IgA、IgG的浓度降低。
Description
技术领域
本发明涉及结肠炎领域，具体涉及一种缓解结肠炎的活性肽VPP及其应用。
背景技术
众所周知，畜禽的健康发展是养殖业经济发展的关键，而肠道健康是影响畜禽健康的关键因素。炎性肠病是一种由肠道生态、遗传、环境等因素引起的肠道免疫疾病。近年来，炎症性肠病在全球范围内呈逐年增加趋势，已成为常见的消化道疾病。炎症反应产生的多种细胞因子和炎性介质会引起组织和肠上皮细胞损伤、肠黏膜屏障功能受损、肠黏膜通透性增加，为致病菌的移位、过度繁殖和转运创造有利条件，最终导致肠道菌群失衡。过度的肠道炎症会诱发肠癌，因此减轻肠道炎症和恢复肠道健康至关重要。
小分子肽Val-Pro-Pro(缬氨酸-脯氨酸-脯氨酸，VPP)是一种乳蛋白来源的生物活性肽。生物活性肽已被证实具有抗癌、降血压、降胆固醇、降血糖、抗氧化、免疫调节和抗菌等多种生理功能。生物活性肽可通过肠上皮细胞吸收而释放到循环血液中，从而使其到达靶器官或系统发挥作用。另外，被吸收的生物活性肽在进入血液循环之前被运输到肝脏进行代谢。生物活性肽由于具有公认的安全性高、成本低和易吸收等多种健康益处。近年来，生物活性肽的抗炎活性也受到越来越多国内外研究学者的关注，因此利用生物活性肽预防和治疗炎症性肠病具有重要意义。
尽管VPP的抗氧化、抑菌性、降血压等活性都有了一定的研究，但其抗炎的作用机制尚不清晰。
发明内容
本发明的目的在于克服现有技术的不足，提供了一种缓解结肠炎的活性肽VPP及其应用。
为实现上述目的，本发明所设计的技术方案如下：
本发明提供了一种缓解结肠炎的活性肽VPP，所述活性肽VPP的氨基酸序列为缬氨酸-脯氨酸-脯氨酸。
本发明还提供了一种上述活性肽VPP在制备预防或治疗结肠炎的药物或食物中的应用。
进一步地，所述结肠炎是DSS诱导的结肠炎。
再进一步地，所述DSS浓度为3％。
本发明还提供了一种上述活性肽VPP在制备治疗仔猪、羔羊或犊牛腹泻的药物中的应用。
本发明还提供了一种预防或治疗结肠炎的产品，所述产品中含有VPP的浓度为10mg/mL。
进一步地，所述产品为药物或食物或饲料。
本发明的有益效果：
(1)本发明所提供的活性肽VPP对动物无毒副作用。
(2)采用本发明所提供的VPP能够显著降低DSS诱导结肠炎期间小鼠的DAI指数，减轻结肠的缩短。
(3)采用本发明所提供的VPP能够缓解DSS诱导的小鼠结肠组织会出现的明显损伤，包括个别肠腺消失被增生的结缔组织取代，杯状细胞数量减少，周围伴有少量的淋巴细胞浸润。
(4)与模型组小鼠相比，VPP干预组小鼠的结肠组织中促炎细胞因子TNF-α、IL-6、IL-8和IL-1β的浓度显著降低。
(5)与模型组小鼠相比，VPP干预使血清免疫球蛋白IgA、IgG的浓度降低。
附图说明
图1为不同组别小鼠体重图、结肠形态图和结肠长度图；
图2为不同组别小鼠结肠组织HE染色图和肠组织损伤评分图；
图3为不同组别小鼠各细胞因子含量图；
具体实施方式
下面结合具体实施例对本发明作进一步的详细描述，以便本领域技术人员理解。
实施例1活性肽VPP合成
缓解结肠炎的活性肽VPP的氨基酸序列为缬氨酸-脯氨酸-脯氨酸，该活性肽VPP按序列由武汉桀升生物科技有限公司合成得到。
实施例2VPP对结肠炎小鼠症状的缓解作用步骤如下：
(1)C57BL/6小鼠(16-18g)30只，随机分为3组，3组分别命名为：空白对照组(Control)、造模组(DSS)、VPP干预组(DSS+VPP)；采用每组10只。
(2)对空白对照组(Control)、造模组(DSS)、VPP干预组(DSS+VPP)进行处理；其中，VPP干预组(DSS+VPP)的处理方法为：实验第1-7天，每天给VPP干预组(DSS+VPP)灌胃VPP10mg/mL 200μL，自由饮用蒸馏水；实验第8-14天，每天给VPP干预组(DSS+VPP)灌胃VPP 200μL和3％DSS溶液200μL，自由饮用3％DSS；
造模组(DSS)的处理方法为：实验第1-7天，每天灌胃200μL生理盐水，自由饮用蒸馏水；实验第8-14天，每天给DSS组灌胃3％DSS溶液200μL,自由饮用3％DSS；
空白对照组(Control)的处理方法为：实验过程中，每天给对照组灌胃生理盐水200μL，自由饮用蒸馏水。
造模期间，检测各组小鼠体重变化。
造模结束小鼠休养三天后，分别将空白对照组(Control)、造模组(DSS)、VPP干预组(DSS+VPP)处死，取整段结肠(盲肠末端至肛门)，测量结肠长度(检测结果见图)并且对结肠外观进行观察。取远端结肠组织1cm进行固定，脱水，包埋，HE染色，观察不同组别小鼠结肠组织HE染色，并评分。
造模结束时，DSS组小鼠的体重显著低于空白对照组(Control)和VPP干预组(DSS+VPP)。
正常组小鼠结肠长度为7.2cm，DSS造模组小鼠的结肠长度仅为4.5cm，而VPP干预组(DSS+VPP)干预显著增加了结肠长度，达到6.5cm(检测结果见图1B和C)。
由图2A可以看出VPP干预组(DSS+VPP)的结肠组织结构与正常组相似，肠组织各层结构清晰；黏膜层上皮完整，上皮细胞形态正常,未见明显的炎性改变。而DSS诱导的小鼠结肠组织出现明显的炎症损伤，包括个别肠腺消失被增生的结缔组织取代，杯状细胞数量轻微减少。VPP干预组(DSS+VPP)的结肠组织损伤评分显著低于DSS组(图2B)。
因此，本发明的VPP可缓解急性结肠炎症状和减轻结肠组织损伤。
实施例3VPP对结肠炎小鼠的免疫调节作用
具体实施方式同实施例1中的步骤(1)-(2)；
将实施例1中的步骤(2)得到的造模结束后小鼠处死，取结肠组织按照结肠组织生化指标测定方法检测结肠组织血清中的生化指标，生化指标包括TNF-α的含量、IL-6的含量、IL-8的含量、IL-1β的含量和血清IgA、IgG的含量。
由图3可知，DSS处理增加了促炎细胞因子TNF-α、IL-6、IL-8和IL-1β的浓度。而VPP处理可以显著降低TNF-α和IL-6、IL-8和IL-1β的浓度。此外，VPP干预使血清免疫球蛋白IgA和IgG的浓度降低。因此，VPP干预后显著改善了肠道的免疫反应，缓解DSS引起的过度免疫系统激活。
其它未详细说明的部分均为现有技术。尽管上述实施例对本发明做出了详尽的描述，但它仅仅是本发明一部分实施例，而不是全部实施例，人们还可以根据本实施例在不经创造性前提下获得其他实施例，这些实施例都属于本发明保护范围。
Claims (7)
1.一种缓解结肠炎的活性肽VPP，其特征在于：所述活性肽VPP的氨基酸序列为缬氨酸-脯氨酸-脯氨酸。
2.一种权利要求1所述活性肽VPP在制备预防或治疗结肠炎的药物或食物中的应用。
3.根据权利要求2所述的应用，其特征在于：所述结肠炎是DSS诱导的结肠炎。
4.根据权利要求3所述的应用，其特征在于：所述DSS浓度为3％。
5.一种权利要求1所述活性肽VPP在制备治疗仔猪、羔羊或犊牛腹泻的药物中的应用。
6.一种预防或治疗结肠炎的产品，其特征在于：所述产品中含有VPP的浓度为10mg/mL。
7.根据权利要求6所述的产品，其特征在于：所述产品为药物或食物或饲料。
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