CN109528704A - 一种增强免疫力的组合物及其在制备免疫增强药物和保健食品中的应用 - Google Patents
一种增强免疫力的组合物及其在制备免疫增强药物和保健食品中的应用 Download PDFInfo
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Abstract
本发明提供了一种增强免疫力的组合物，属于免疫调节药物技术领域，所述组合物包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.4～0.6)∶(0.8～1.2)∶(17～25)。所述组合物能够提高免疫抑制小鼠的脾脏指数和胸腺指数；提高白细胞总数，廓清指数K以及吞噬指数α；能够使血清中IL‑2、TNF‑α和IFN‑γ的含量水平趋近于正常水平，具有显著的增强免疫力的作用。
Description
技术领域
本发明属于免疫调节药物技术领域，尤其涉及一种增强免疫力的组合物及其在制备免疫增强药物和保健食品中的应用。
背景技术
大蒜(Allium sativum L.)是葱属植物大蒜的鳞茎，有着悠久的历史，是人类种植的最古老的具有药效的植物，也是药食两用植物，含有氨基酸类、糖类、苷类及含硫化合物等多种化学成分，其含量随品种、种植环境、采收和贮藏时间不同而异。大蒜化学成分的多样性是大蒜防病治病的基础，具有抗氧化、抗肿瘤、抗病原微生物、抗心肌缺血、降血脂、免疫调节等多种药理作用，主要用于抗炎、抗癌、心脑血管疾病和糖尿病，作为动物饲料添加剂可改变肌肉组织化学成分，影响血浆相关参数，有效降低死亡率。
现有研究多集中在大蒜水提物或乙醇提取物、大蒜辣素的降解产物大蒜素和大蒜油、提取的大蒜多糖等。Salman H(Bessler H，Bergman C M，Salmanroghani H，etal.Effect of a garlic derivative(alliin)on peripheral blood cell immuneresponses.[J].International Journal of Immunopharmacology，1999，21(9)：589.)通过离体实验表明，蒜氨酸可以促进美洲商陆诱导的外周血单核细胞增生，IL-1β和TNF-α产生，以及吞噬细胞吞噬能力的提高。王钦富(王钦富，陈玉良.口服大蒜匀浆对小鼠免疫功能的影响[J].大连大学学报，1993(3)：147-148.)等以大蒜匀浆喂饲小鼠1个月后，测得淋巴细胞百分率、白细胞总数和ANAE阳性细胞数都极显著高于对照组，结果表明大蒜能明显提高小鼠特异性细胞免疫功能。李锋(李锋，王玉.大蒜浸提液对小鼠细胞免疫功能影响研究[J].卫生研究，1995(4)：250-251.)等人研究认为：大蒜浸提液可使小鼠胸腺和脾脏重量及脏器系数增加，同时明显提高淋巴细胞转化率及T淋巴细胞酸性α-醋酸萘酯酶(ANAE)阳性率。岳丽(岳丽，王辉，黄雪松，等.大蒜多糖B对免疫抑制小鼠免疫活性的调节[J].暨南大学学报：医学版，2009，30(6)：601-605.)等的研究表明大蒜粗多糖可能通过影响巨噬细胞对抗原处理的能力以及T淋巴细胞的分化成熟来调节免疫抑制小鼠的机体免疫活性。
但现有研究中并不清楚发挥提高免疫功效的具体成分，尚没有一种成分明确的具有增强免疫力的组合物。
发明内容
有鉴于此，本发明的目的在于提供一种增强免疫力的组合物及其在制备免疫增强药物和保健食品中的应用，所述组合物能够提高免疫抑制小鼠的脾脏指数和胸腺指数；提高白细胞总数，廓清指数K以及吞噬指数α；能够使血清中IL-2、TNF-α和IFN-γ的含量水平趋近于正常水平，具有显著的增强免疫力的作用。
为了实现上述发明目的，本发明提供了以下技术方案：
一种增强免疫力的组合物，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.4～0.6)∶(0.8～1.2)∶(17～25)。
优选的，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.45～0.55)∶(0.9～1.1)∶(19～23)。
优选的，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为0.5∶1.0∶(20～22)。
本发明还提供了所述的组合物在制备免疫增强药物中的应用。
优选的，所述免疫增强药物为口服制剂。
优选的，所述免疫增强的表现包括脾脏指数和胸腺指数的提高。
优选的，所述免疫增强的表现包括提高白细胞总数的提高、廓清指数K以及吞噬指数α的提高。
优选的，所述免疫增强的表现包括血清中IL-2、TNF-α和IFN-γ含量的提高。
本发明还提供了所述组合物在制备保健食品中的应用。
本发明的有益效果：本发明提供的增强免疫力的组合物，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.4～0.6)∶(0.8～1.2)∶(17～25)。本发明中所述蒜氨酸和大蒜多糖均具有增强免疫力的作用，所述大蒜凝集素作为一种增强剂，能够增强所述蒜氨酸和大蒜多糖的作用；将所述大蒜凝集素、蒜氨酸和大蒜多糖三者按照上述比例混合获得的组合物，能够提高免疫抑制小鼠的脾脏指数和胸腺指数；提高白细胞总数，廓清指数K以及吞噬指数α；并且能够使免疫抑制小鼠血清中IL-2、TNF-α和IFN-γ的含量水平趋近于正常水平，具有显著的免疫增强活性，有良好的开发应用潜力。
具体实施方式
本发明提供了一种增强免疫力的组合物，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.4～0.6)∶(0.8～1.2)∶(17～25)，优选为(0.45～0.55)∶(0.9～1.1)∶(19～23)，更优选为0.5∶1.0∶(20～22)。
在本发明中，所述增强免疫力的组合物包括大蒜多糖，本发明对所述大蒜多糖的来源没有特殊限定，采用市售大蒜多糖或自制大蒜多糖均可。本发明中，所述自制大蒜多糖的方法包括以下步骤：鲜蒜微波灭酶、打浆、分离得到上清液，采用离子交换树脂进行吸附后，收集水洗部分，浓缩干燥后即得。本发明中，所述大蒜多糖具有提高免疫抑制小鼠的脾脏指数和胸腺指数；提高白细胞总数，廓清指数K以及吞噬指数α的作用；并且能够使免疫抑制小鼠血清中IL-2、TNF-α和IFN-γ的含量水平趋近于正常水平。
在本发明中，所述增强免疫力的组合物包括蒜氨酸，本发明对所述蒜氨酸的来源没有特殊要求，采用市售蒜氨酸或自制蒜氨酸均可。所述自制蒜氨酸的方法参考中国专利ZL 03100420.2“从鲜蒜中提取蒜氨酸的生产工艺”。本发明中，所述蒜氨酸能够提高免疫抑制小鼠的吞噬指数α，提高TNF-α、IFN-γ含量水平。
在本发明中，所述增强免疫力的组合物包括大蒜凝集素，本发明中，所述大蒜凝集素为从大蒜中提取获得的凝集素，为甘露糖结合凝集素。在本发明具体实施过程中，所述大蒜凝集素为鲜蒜匀浆破碎后取上清液，过葡聚糖凝胶柱后收集相应大蒜凝集素组分。本发明中，所述大蒜凝集素作为增强剂，能够增强大蒜多糖和蒜氨酸的提高免疫力相关的作用。
本发明中所述组合物的制备方法为将上述大蒜凝集素、蒜氨酸和大蒜多糖按照比例混合。本发明中，所述大蒜凝集素、蒜氨酸和大蒜多糖按照上述比例混合后，三者之间能够发挥协同增效作用，使所述组合物能够显著提高机体免疫力。
本发明还提供了所述的组合物在制备免疫增强药物中的应用。在本发明中，所述免疫增强药物优选为口服制剂，本发明对所述口服制剂的具体剂型没有特殊限定，采用本领域常规口服制剂的剂型即可。本发明中，所述免疫增强的表现包括脾脏指数和胸腺指数的提高、提高白细胞总数的提高、廓清指数K以及吞噬指数α的提高以及血清中IL-2、TNF-α和IFN-γ含量的提高。
本发明还提供了所述组合物在制备保健食品中的应用。本发明对所述保健食品的类型没有特殊限定，采用本领域常规保健食品的类型即可。本发明中，所述保健食品能够增强免疫力。
下面结合实施例对本发明提供的技术方案进行详细的说明，但是不能把它们理解为对本发明保护范围的限定。
实施例1
一种增强免疫力的组合物，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为0.5∶1.0∶20。将所述大蒜凝集素、蒜氨酸和大蒜多糖进行物理混合获得组合物。
实施例2
一种增强免疫力的组合物，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为0.6∶0.9∶22。将所述大蒜凝集素、蒜氨酸和大蒜多糖进行物理混合获得组合物。
实施例3
一种增强免疫力的组合物，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为0.4∶1.1∶25。将所述大蒜凝集素、蒜氨酸和大蒜多糖进行物理混合获得组合物。
实施例4
组合物对免疫抑制小鼠免疫器官的影响
192只小鼠随机平均分为8组，设置正常对照组、模型组、阳性药物组，组合物高剂量组、组合物低剂量组、组合物中剂量组、大蒜多糖组、大蒜多糖+蒜氨酸组。
正常对照组：不进行任何处理。
模型组：选用腹腔注射环磷酰胺(CTX)制造免疫抑制小鼠模型，具体的0.1ml/10g的剂量进行注射，使所述小鼠体内环磷酰胺的浓度达到40mg/Kg；连续7天注射阳性药物，饮食和照料正常进行。
阳性药物组：以左旋咪唑(LMS)作为阳性药物对照，注射方法与剂量和模型组一致，饮食和照料正常进行。
组合物组：大蒜凝集素、蒜氨酸和大蒜多糖的质量比为0.5∶1.0∶21，其中高中低三个剂量组以蒜氨酸量计。
组合物高剂量组：以60mg/kg的浓度灌胃30d；
组合物中剂量组：以17.5mg/kg的浓度灌胃30d；
组合物低剂量组：以5mg/kg的浓度灌胃30d；
大蒜多糖组：以164mg/kg的浓度灌胃30d；
大蒜多糖+蒜氨酸组(质量比1∶20)：以164mg/kg的浓度灌胃30d。
实验结果：由表1可知，模型组的脾脏指数和胸腺指数明显低于正常对照组，差异有统计学意义(P＜0.05)，造模成功。
与模型组比较，各给药组均可提高环磷酰胺降低的脾脏指数(P＜0.05或P＜0.01)。阳性药组、大蒜多糖组、组合物各剂量组均可提高环磷酰胺降低的胸腺指数(P＜0.05或P＜0.01)。可见各处理组对环磷酰胺所致的脾脏指数和胸腺指数都有一定的恢复功能。结果详见表1。
表1.各处理组对免疫抑制小鼠脾脏指数和胸腺指数的影响(x±s，n＝12)
注：与正常对照组比较，*P＜0.05，**P＜0.01；与CTX模型组比较，ΔP＜0.05，ΔΔP＜0.01。
2各处理组对免疫抑制小鼠外周血白细胞数的影响
由表2可知，模型组的白细胞数明显低于正常对照组，差异有统计学意义(P＜0.05)，造模成功。
与模型组比较，阳性药组、大蒜多糖组、组合物各剂量组均可提高环磷酰胺降低的白细胞数(P＜0.05或P＜0.01)，可见各处理组对环磷酰胺所致的外周血白细胞减少都有恢复功能，且具有正向免疫调节作用。
表2.各处理组对免疫抑制小鼠外周血白细胞数的影响(x±s，n＝12)
注：与正常对照组比较，*P＜0.05，**P＜0.01；与CTX模型组比较，ΔP＜0.05，ΔΔP＜0.01。
3各处理组对免疫抑制小鼠血清IL-6、TNF-α和IFN-γ含量的影响
从表3可以看出，模型组小鼠的IL-6、TNF-α、IFN-γ含量都明显低于正常对照组，差异有统计学意义(P＜0.05或P＜0.01)，造模成功。
与模型组比较，阳性药组、大蒜多糖组、组合物高中剂量组均可明显恢复环磷酰胺降低的IL-6含量水平，各给药组均可恢复环磷酰胺降低的TNF-α、IFN-γ含量水平。差异有统计学意义(P＜0.05或P＜0.01)。大蒜凝集素+蒜氨酸+大蒜多糖各剂量组之间差异不显著。
表3各处理组对免疫抑制小鼠血清IL-6、TNF-α和IFN-γ含量的影响(x±s，n＝12)
注：与正常对照组比较，*P＜0.05，**P＜0.01；与CTX模型组比较，ΔP＜0.05，ΔΔP＜0.01。
4化学成分(组)对免疫抑制小鼠巨噬细胞吞噬功能的影响
从表4以看出，模型组的廓清指数K和吞噬指数α均明显低于正常对照组，差异有统计学意义(P＜0.05)，造模成功。
与模型组比较，组合物中低剂量组和蒜氨酸+大蒜多糖组可明显恢复环磷酰胺降低的免疫抑制小鼠的吞噬指数α(P＜0.05或P＜0.01)。组合物各剂量组之间差异不显著(P＞0.05)。
表4各处理组对免疫抑制小鼠巨噬细胞吞噬功能的影响(x±s，n＝12)
注：与正常对照组比较，*P＜0.05，**P＜0.01；与CTX模型组比较，ΔP＜0.05，ΔΔP＜0.01。
本实验将小鼠连续30天灌胃给予大蒜多糖、组合物和蒜氨酸+大蒜多糖，结果显示，组合物中低剂量组和蒜氨酸+大蒜多糖组可明显恢复造模降低的免疫抑制小鼠的吞噬指数α；大蒜多糖组、组合物高中剂量组可恢复因造模降低的IL-6含量水平；各给药组均可恢复降低的TNF-α、IFN-γ含量水平。(P＜0.05)。各给药组的脾脏指数、胸腺指数、白细胞计数、吞噬指数、IL-6、IFN-γ、TNF-α含量水平恢复效果优于阳性药组。同时，仅有大蒜多糖单一成分的效果不如蒜氨酸+大蒜多糖协同作用效果好，而大蒜凝集素+蒜氨酸+大蒜多糖的协同效果又优于蒜氨酸+大蒜多糖组。
本发明所述组合物能够升高免疫抑制小鼠的免疫因子浓度，对免疫抑制小鼠的免疫功能具有正向调节作用，但无剂量依赖关系，在可提高机体免疫力水平的前提下，组合物的协同效果最为显著。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (9)
1.一种增强免疫力的组合物，其特征在于，包括大蒜凝集素、蒜氨酸和大蒜多糖，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.4～0.6)∶(0.8～1.2)∶(17～25)。
2.根据权利要求1所述的组合物，其特征在于，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为(0.45～0.55)∶(0.9～1.1)∶(19～23)。
3.根据权利要求1或2所述的组合物，其特征在于，所述大蒜凝集素、蒜氨酸和大蒜多糖的质量比为0.5∶1.0∶(20～22)。
4.权利要求1～3任意一项所述的组合物在制备免疫增强药物中的应用。
5.根据权利要求4所述的应用，其特征在于，所述免疫增强药物为口服制剂。
6.根据权利要求4所述的应用，其特征在于，所述免疫增强的表现包括脾脏指数和胸腺指数的提高。
7.根据权利要求4或6所述的应用，其特征在于，所述免疫增强的表现包括白细胞总数的提高、提高廓清指数K以及吞噬指数α的提高。
8.根据权利要求7所述的应用，其特征在于，所述免疫增强的表现包括血清中IL-2、TNF-α和IFN-γ含量的提高。
9.权利要求1～3任意一项所述的组合物在制备保健食品中的应用。
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