CN1889853B - 包含谷氨酰胺的抗腹泻组合物 - Google Patents

包含谷氨酰胺的抗腹泻组合物 Download PDF

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CN1889853B
CN1889853B CN2004800360410A CN200480036041A CN1889853B CN 1889853 B CN1889853 B CN 1889853B CN 2004800360410 A CN2004800360410 A CN 2004800360410A CN 200480036041 A CN200480036041 A CN 200480036041A CN 1889853 B CN1889853 B CN 1889853B
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C·库
K·L·格罗斯
D·杰维尔
K·维德金德
S·兹克
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Abstract

本发明提供了在患有胃肠道炎症的哺乳动物中适合于哺乳动物口服摄取的组合物，其包括抗腹泻有效量的谷氨酰胺、发酵性纤维、抗氧化剂和ω-3脂肪酸。

Description

包含谷氨酰胺的抗腹泻组合物
发明背景
保持哺乳动物胃肠道的状态良好是非常令人期望的目标。特别的烦恼在于胃肠道的炎性状况。胃肠道炎症的一些征候包括急性或慢性腹泻、软便、便血、呕吐、营养物质消化和吸收差、体重减轻和食欲不振。已知疾病如胃炎、肠炎、结肠炎、炎症性肠病、溃疡、某些类型的癌症、和已知的以GI炎症作为其主要构成的其它病症。
我们发现某些物质的混合物可以带来GI炎症的基本征候如腹泻的改善。当对哺乳动物口服给用适当量的谷氨酰胺、发酵性纤维、抗氧化剂和ω(n)-3脂肪酸时，在胃肠道炎症得到改善的同时，排便频率以及排便的质量可以得到显著改善，特别是在伴侣宠物如猫中。
发明简述
根据本发明，提供了在患有胃肠道炎症的哺乳动物中适合于哺乳动物口服摄取的组合物，该组合物包括抗腹泻有效量的谷氨酰胺、发酵性纤维、抗氧化剂和ω-3脂肪酸的组合。
本发明的另一个方面是处理患有胃肠道炎症的哺乳动物中的腹泻的方法，包括对哺乳动物口服给用上述组合物。
发明详述
公知谷氨酰胺对于淋巴细胞的增殖是重要物质并且是肠细胞的重要营养物质。谷氨酰胺还是谷胱甘肽的前体，谷胱甘肽为体内的天然抗氧化剂。本文中公开的用于任何组分的所有重量％都基于哺乳动物的日常膳食。所有的数字都以干物质组成计算。
谷氨酰胺的量最低为约0.1、0.15或0.2重量％。最大量通常不超过约5、4或3重量％。
可以使用的纤维是具有适度发酵性的纤维、高度发酵性的纤维、或为两者的混合物。也可在不影响制剂效果的低水平下加入低发酵性或非发酵性的纤维。
我们已经表明，某些益生纤维组分当被狗和猫胃肠道中存在的细菌发酵时产生大量的丁酸脂和其它短链脂肪酸，其将酸化胃肠道并降低病原体的生长。产生大量丁酸酯的益生纤维包括但不限于甘露寡聚糖、胶质、寡木糖、牛蒡、甜菜浆、菊粉、半乳糖、其它木聚糖、果聚糖、右旋糖酐、β葡聚糖、抗性淀粉、得自胶类的多糖等，在膳食中应当以约0.5-20重量％的量存在，优选为约1-5重量％。胶类可包括由微生物产生的胶类如洁冷胶、黄原胶或由植物产生的胶类如阿拉伯树胶。混合物的配制优选基于高的丁酸酯生成和基于挥发性脂肪酸(VFA)生成的中等发酵能力和有机物质消失率，以帮助保持最佳的GI健康。组合物可以包括至少约10-60％的适度发酵性纤维和约20-40％的高度发酵性纤维。这些纤维应选择为使得这些纤维的丁酸酯生成量较高，为总VFA的约5-40％。适度发酵性纤维的定义为当由粪便细菌体外发酵24小时周期时有机物质消失率为约15到60％。也就是说，最初存在的总有机物质的约15到60％被粪便细菌发酵和被转换掉。高度发酵性纤维具有大于60％的消失率。
在组合物和方法中还可使用抗氧化剂。可以使用维生素E、C及其掺混物。可使用这些维生素的任何前体，诸如醋酸生育酚和抗坏血酸钠。维生素E最低为膳食的约0.1、0.2或0.4重量％，通常最大不超过膳食的约3、2或1重量％。维生素C最低为膳食的约0.1、0.2或0.4重量％，通常最大不超过膳食的约3、2或1重量％。
ω-3脂肪酸是公知的膳食组分，主要在油类和脂类中发现，特别是在鱼油诸如鲜鱼、鲑鱼等鱼油中。ω-3脂肪酸的基本组分为二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)和α-亚麻酸(ALA)。ω-3脂肪酸的量通常为最低约0.1、0.2或0.5重量％，通常最大不超过约3、2和1重量％。通常还存在于油类和脂类中的是ω-6脂肪酸。当存在时，ω-6脂肪酸对ω-3脂肪酸的比以重量计为约0.5∶1到6∶1，优选约2∶1到4∶1。
以下实施例说明使用本发明的组合物在处置哺乳动物的腹泻时实现的益处。哺乳动物患有或可患有胃肠道炎症(优选炎症性肠病)。
实施例1
在以下研究中，用2种食品喂养患有炎症性肠病(IBD)的12只猫，各自喂养2周。对6只猫喂养食品A2周，对6只猫喂养食品B2周，之后交叉喂养。每日监控粪便质量，并基于1-5等级进行评分，1分为过稀且多水，5分为坚硬且成形。评分如下。得自患有IBD的猫的粪便通常为1或2分。
粪便监控评分
1：多水
2：软，未成形
3：软，成形，湿润
4：坚硬，成形，干燥
5：坚硬，干燥小球
表1表明膳食对患有慢性腹泻的猫的粪便质量的作用。表1表示获得1-5分的粪便百分数。第一罐装食品A包含3％的低发酵能力(低于15％)的纤维，罐装食品B包含1.5％的高发酵能力(高于约60％)的纤维，食品的营养含量如下所示。
  食品A   食品B
  低发酵性纤维食品   高发酵性纤维食品
  湿度   72.69   72.58
  蛋白质-凯氏法   8.24   7.94
  粗纤维   0.3   0.2
  酸解的粗脂肪   9.58   9.85
结果
结果表明，喂养含有高发酵性纤维源的食品B改善猫的粪便质量，从42％的粪便百分数(得分为1和2分)改善到仅仅15％的粪便百分数(得分为1和2分)。
表1
Figure B2004800360410D00041
＊4％的粪便不能用于等级评价
实施例2
表2表示实施例1中的相同猫用2种不同食品喂养进行研究的数据。两种食品都含有类似量的益生纤维和ω-3脂肪酸。食品C含有额外的谷氨酰胺和抗氧化剂，而食品D不含有额外的谷氨酰胺或抗氧化剂。半数猫喂养食品C2周，另外半数猫喂养食品D，然后所有猫有1周的冲洗期。然后交叉喂养另一种食品，再喂养2周。表2的结果表明，与当对猫喂养食品D(该膳食没有额外的谷氨酰胺和抗氧化剂)的粪便质量相比(得分为1和2的粪便百分数为7％)，当对猫喂养含有谷氨酰胺和高抗氧化剂的食品C时，粪便质量有显著改善(得分为1和2的粪便百分数为0％)。与食品A(得分为1和2的粪便百分数为42％)相比，食品C的粪便质量具有显著更好的结果，得分为1和2的粪便百分数为0％。食品C还显著好于食品B(得分为1和2的粪便百分数为15％)。食品C还显著好于食品D(得分为1和2的粪便百分数为7％)。食品C具有本发明所有的显著组分：谷氨酰胺、抗氧化剂、发酵性纤维和n-3脂肪酸。食品A、B和D都缺少这些组分中的至少一种。
食品的营养含量如下所示。
Figure B2004800360410D00051
表2
粪便百分数
Figure B2004800360410D00052
数据表明具有额外的谷氨酰胺和抗氧化剂的膳食在这些猫中持续维持粪便质量的改善。
实施例3
以下实验表明在上述实施例中使用的谷氨酰胺源具有生物利用度，并且能够激发免疫功能。谷氨酰胺是肠道的重要营养物质，因为其是肠细胞和淋巴细胞的重要能量来源。膳食中的大部分谷氨酰胺被肠细胞以及肠中的免疫细胞所吸收。
在一个实验中，试验了谷氨酰胺的来源，看看其是否具有生物利用度并且能够递送足够的谷氨酰胺到肠细胞。谷氨酰胺的来源是小麦水解产物，其富集30％的谷氨酰胺。在6只狗中进行剂量反应研究，看看在喂养膳食后在血浆中是否可检测到增加量的谷氨酰胺源(0、0.5、1.0、2％的谷氨酰胺含量)。
表3
在用增补有不同量的谷氨酰胺的食品喂养的动物中，饭后血浆谷氨酰胺的变化
  增补的谷氨酰胺％   饭后血浆谷氨酰胺相对于对照的变化％
  0.5％   3
  1.0％   10
  2.0％   15
数据表明对膳食中增加量的谷氨酰胺有增加的反应，特别是在饭后30分钟。由此可见，谷氨酰胺在被肠细胞摄取后可被血流获得。
在另一个实验中，检测了谷氨酰胺作为免疫调节剂的效力。随机将20只小猎犬分成4组，它们接受基本膳食或增补有1％、2％或者4％谷氨酰胺的基本膳食。在第1天和第16天，在动物停食后2小时抽取血样在肝素化试管中，样品用于免疫测量(T细胞增生试验)。
T细胞增殖试验。使用Nova Celltrak II(Beckman Coulter Corp.，FL)计数每个血样中的外周血液白细胞(PBL)。血液用补给性培养基稀释(1∶20)，将稀释后的血液涂板在96孔细胞培养物板中，一式三份，细胞培养物板具有在补给性培养基中稀释的以下促分裂素：伴刀豆球蛋白A(0.5μg/ml，2.5μg/ml，和10μg/ml)、PWM(1.5μg/ml、2.5μg/ml)、和PHA(0.5μg/ml、2.5μg/ml)。将板在包含7％CO2的湿润培养箱中在37℃培养72小时。细胞DNA为Ci/孔[在用13H收集之前脉冲标记18小时]胸苷。细胞DNA在玻璃纤维纸上使用细胞收集器(Skatron Instruments Inc.，VA)收集并使用1.5毫升闪烁鸡尾酒悬浮。[3H]胸苷吸收使用TriCarb 2100TR液体闪烁分析器(Packard BioScience Company，IL)被定量为每分钟计数(CPM)，计数归一化为CPM/10,000细胞，以说明PBL浓度的变化。
谷氨酰胺对淋巴细胞增殖的作用
伴刀豆球蛋白A(Con A)为多克隆T细胞促分裂素。在Con A促分裂素的存在下，全面分析表明膳食(无Con A剂量的效应)、或由Con A相互作用的膳食处置的显著作用。因此，在不同剂量Con A范围内压缩数据以表明随着膳食中谷氨酰胺的增补百分数的不同而变化的淋巴细胞的增殖反应。
表4：T细胞淋巴细胞响应Con A促分裂素的增殖
  食品   T细胞增殖(log<sub>10</sub>cpm)
  未增补谷氨酰胺   4.7
  增补1％的谷氨酰胺   5
  食品   T细胞增殖(log<sub>10</sub>cpm)
  增补2％的谷氨酰胺   4.8
  增补4％的谷氨酰胺   4.5
膳食处置有显著性主要效应(P＜0.01)。增补1％谷氨酰胺的膳食表现出最大的淋巴细胞增殖，其显著不同于对照组(p＜0.05)。供给1％和2％谷氨酰胺的狗表现出淋巴细胞增殖的类似增加。在这些组和其膳食中增补有4％谷氨酰胺的动物中的淋巴细胞的增殖反应之间具有显著性差异(P＜0.01)，由此表明增补1-2％的谷氨酰胺增强总的T淋巴细胞增殖。然而，4％谷氨酰胺在这方面没有附加的益处。

Claims (14)

1.在患有胃肠道炎症的哺乳动物中适合于哺乳动物口服摄取的组合物，其包括抗腹泻有效量的谷氨酰胺、发酵性纤维、抗氧化剂和ω-3脂肪酸。
2.权利要求1的组合物，其中哺乳动物为狗或猫。
3.权利要求2的组合物，其中组合物以狗或猫的膳食形式被给用。
4.权利要求3的组合物，其中谷氨酰胺为膳食的约0.1到约5重量％。
5.权利要求3的组合物，其中发酵性纤维为膳食的约0.5到约20重量％。
6.权利要求3的组合物，其中抗氧化剂为膳食的约0.1到约3重量％。
7.权利要求3的组合物，其中ω-3脂肪酸为膳食的约0.1到约3重量％。
8.权利要求4的组合物，其中发酵性纤维为膳食的约0.5到约20重量％。
9.权利要求4的组合物，其中抗氧化剂为膳食的约0.1到约3重量％。
10.权利要求4的组合物，其中ω-3脂肪酸为膳食的约0.1到约3重量％。
11.权利要求8的组合物，其中抗氧化剂为膳食的约0.1到约3重量％。
12.权利要求8的组合物，其中ω-3脂肪酸为膳食的约0.1到约3重量％。
13.权利要求11的组合物，其中ω-3脂肪酸为膳食的约0.1到约3重量％。
14.权利要求1的组合物在制备用于处置患有胃肠道炎症的哺乳动物中的腹泻的制品中的用途。
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