CN116235948A - 一种抗糖化组合物及其应用 - Google Patents
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Abstract
本发明提供一种抗糖化组合物及其应用。该抗糖化组合物包括以下重量份的组分:含多酚的山竹提取物0.5‑5份、含茶多酚的绿茶提取物0.5‑5份、含多酚的线叶金雀花提取物1份。该组合物中包括含多酚的山竹提取物、含茶多酚的绿茶提取物、含多酚的线叶金雀花提取物,三者之间具有良好的协同增效作用,使得该组合物具有良好的抗糖化功效,还兼具抗氧化功效。利用该组合物制备得到的食品、保健品具有抗糖化和抗氧化功效。
Description
技术领域
本发明涉及植物提取物领域,具体涉及一种抗糖化组合物及其应用。
背景技术
皮肤衰老主要分为内源性衰老和外源性衰老两种形式。内源性衰老是由机体的遗传因素决定的,是自然发生的过程,难以预防,主要表现为皮肤干燥、松弛、细纹、下垂、皮肤变薄、皮下脂肪减少(引起眼窝和颊部凹陷等)。外源性衰老是由环境因素(紫外线、尼古丁、空气污染、微生物等)或者生活方式产生的皮肤积累性损害造成的。外源性衰老不具有普遍性,其在一定程度上可以预防,主要发生在皮肤暴露的部位,如面部、颈部、手背和前臂等,具体表现为较深的皱纹、老年斑和毛细血管扩张等。
非酶糖基化(Non-enzamatic glysation,NEG)是指蛋白质、脂类、核酸等生物大分子在非酶条件下,自发地与葡萄糖或其他还原单糖发生一连串反应,产生一种叫晚期糖化终产物(advanced glycation end products,AGEs)的物质的过程。非酶糖基化是外源性衰老的一个重要因素,其在生物体内广泛而缓慢地进行着。AGEs的形成是一个复杂的不可逆的多步骤过程,羧甲基赖氨酸(CML)和戊糖苷是皮肤中最普遍的AGEs。随着年龄的增长,AGEs在体内不断积累,能使相邻的蛋白质、糖类、脂类、核酸等物质发生交联,不仅影响这些物质的结构,也可造成其生物学性能的改变,在衰老进程中扮演着重要的角色。通常所说的“抗糖化”其实就是抗AGEs。
有研究发现AGEs与细胞外基质蛋白如胶原蛋白,弹性蛋白有关。胶原蛋白I(真皮中的主要结构蛋白之一)的非酶糖基化以多种方式损害其功能。据报道,AGEs修饰的胶原蛋白和弹性蛋白具有修饰的生物力学特性,从而导致弹性丧失和僵硬的增强,促进皱纹的出现。NEG还可以改变胶原蛋白与细胞的相互作用,从而影响其功能,例如迁移,分化和增殖。在光老化的皮肤中存在非酶糖基化的弹性蛋白,这表明紫外线辐射刺激了弹性蛋白的非酶糖基化。另外,非酶糖基化的细胞外基质蛋白似乎对基质金属蛋白酶的降解更有抵抗力,因此减慢了其被新合成和功能蛋白的去除和替代。
发明内容
本发明的目的在于提供一种抗糖化组合物及其应用,以抑制生物体内的非酶糖基化反应,进而延缓衰老进程。
根据本发明的第一个方面,提供一种抗糖化组合物,该抗糖化组合物包括以下重量份的组分:含多酚的山竹提取物0.5-5份、含茶多酚的绿茶提取物0.5-5份、含多酚的线叶金雀花提取物1份。
山竹的学名为山莽吉柿,俗称山竺、山竹子、倒捻子,属于藤黄科藤黄属,具有极高的营养价值,中医认为有清热降火、美容肌肤的功效。山竹中含有丰富的山酮素,具有抗自由基作用。目前所知的200多种山酮素,山竹中就有40多种。此外,山竹中含有的表儿茶素、多酚等能够清除过多的自由基,增强抗皱功效,延缓衰老。
绿茶中的茶多酚具有很强的抗氧化和生理活性,是一种高效的人体自由基清除剂,其清除自由基的能力大大高于其他同类物质;茶多酚还具有阻断脂质过氧化反应,清除活性酶的作用,其抗衰老效果显著强于维生素E。
线叶金雀花是南非赛德堡地区生长的一种草本针状灌木,其富含多种黄酮类物质和多酚,具有极强的抗氧化能力。此外,线叶金雀花中含有丰富的超氧化物歧化酶(SOD),能够促进胶原蛋白、弹性蛋白的合成,导致体内活性氧等氧化自由基减少,减缓氧化作用,下调基质金属蛋白酶的活性以预防紫外线对皮肤的损伤,同时有利于改善肤质,具有一定的美容效果,有助于延缓衰老。
本发明提供的抗糖化组合物中含有含多酚的山竹提取物、含茶多酚的绿茶提取物、含多酚的线叶金雀花提取物,这三种提取物之间具有良好的协同增效作用,把这三种提取物进行组合,同时通过控制各组分的含量,发现该组合物具有显著的抗糖化功效,同时还兼具抗氧化作用。
优选地,上述抗糖化组合物包括以下重量份的组分:含多酚的山竹提取物1-2.5份、含茶多酚的绿茶提取物1-2.5份、含多酚的线叶金雀花提取物1份。
优选地,含多酚的山竹提取物中多酚含量≥0.01%,含茶多酚的绿茶提取物中茶多酚含量≥50%,含多酚的线叶金雀花提取物中多酚含量≥15%。
优选地,上述含多酚的山竹提取物由以下步骤制备得到:将山竹加入溶剂进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到含多酚的山竹提取物。
优选地,上述含茶多酚的绿茶提取物由以下步骤制备得到:将绿茶加入溶剂进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到含茶多酚的绿茶提取物。
优选地,上述含多酚的线叶金雀花提取物由以下步骤制备得到:将线叶金雀花加入溶剂进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到含多酚的线叶金雀花提取物。
优选地,上述溶剂均为水。
根据本发明的第三个方面,提供抗糖化组合物在制备抗糖化食品、抗糖化保健品中的应用。
根据本发明的第四个方面,提供一种抗糖化食品,包括上述抗糖化组合物,抗糖化组合物在抗糖化食品中的重量占比为0.01-25%。
优选地,上述抗糖化食品还包括食品领域可接受的辅料或添加剂,辅料或添加剂包括甜味剂、酸调节剂、填充剂、矫味剂、着色剂、抗氧化剂、增稠剂、稳定剂、乳化剂、抗结剂、助流剂、润滑剂中的至少一种。
优选地,抗糖化食品包括饮品、果冻、固体饮料、压片糖果、软糖。
根据本发明的第五个方面,提供一种抗糖化保健品,该抗糖化保健品包括上述抗糖化组合物,抗糖化组合物在抗糖化保健品中的重量占比为0.01-25%。
优选地,上述抗糖化保健品还包括保健品领域可接受的辅料或添加剂,辅料或添加剂包括甜味剂、酸调节剂、填充剂、矫味剂、着色剂、抗氧化剂、增稠剂、稳定剂、乳化剂、抗结剂、助流剂、润滑剂中的至少一种。
优选地,抗糖化保健品包括口服液、胶囊、片剂、粉剂。
利用上述抗糖化组合物制备抗糖化食品、抗糖化保健品,使得该食品、保健品具有优异的抗糖化功效,同时具有很强的抗氧化作用,
本发明的有益效果为:
1.本发明利用含多酚的山竹提取物、含茶多酚的绿茶提取物、含多酚的线叶金雀花提取物制备抗糖化组合物,这三种提取物之间具有协同增效作用,使得该组合物具有良好的抗糖化功效,还兼具抗氧化效果。
2.将上述抗糖化组合物应用于抗糖化食品、抗糖化保健品的制备中,使其具有抗糖化以及抗氧化功效。
具体实施方式
下面结合具体实施方式对本发明提供的技术方案中的技术特征作进一步清楚、完整的描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1-5和对比例1-6
一种抗糖化组合物,其制备方法包括以下步骤:
(1)将山竹加入水中进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到含多酚的山竹提取物;
(2)将绿茶加入水中进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到含茶多酚的绿茶提取物;
(3)将线叶金雀花加入水中进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到含多酚的线叶金雀花提取物;
(4)将步骤(1)得到的含多酚的山竹提取物0.5-5份、步骤(2)得到的含茶多酚的绿茶提取物0.5-5份、步骤(3)得到的含多酚的线叶金雀花提取物1份混合均匀,得到抗糖化组合物(表1)。
表1抗糖化组合物组分
测试例
1.实验构建方式
(1)非酶糖基化产物抑制实验
本实验所采用的参试对象为实施例1-5所制得的抗糖化组合物。采用荧光分析法测定AGEs的生成量,对组合物的抗糖化作用进行评价。组合物对非酶糖基化产物的抑制强度用抑制率表示。具体实验步骤如下:
①分别用磷酸盐缓冲液(PBS缓冲液,50mmol/L,pH7.4,含有0.04%Proclin200)配制30mg/mL的BSA溶液(牛血清白蛋白溶液)、1.5mol/L的果糖溶液、不同质量浓度的实验样品;
②向5mL EP管中加入1mL果糖溶液和1mL不同浓度的样品溶液作为样品组,向5mLEP管中加入1mL果糖溶液和1mL PBS缓冲液作为阴性对照组,密封好EP管,在37℃下孵育2h,向样品组和阴性对照组中分别加入1mL BSA溶液,在37℃下孵育6d。
③AGEs荧光产物的测定:从每组转移200μL混合液至96孔板中,以PBS缓冲液作为空白对照用酶标仪在激发波长为370nm、发射波长420nm条件下测定荧光强度。样品对AGEs生成的抑制率(%)计算公式如下:AGEs抑制率(%)=[1-(F样品/F阴性对照)]×100%,其中,式中的F样品表示样品组测得的荧光强度,F阴性对照表示阴性对照组测得的荧光强度。
表2抗糖化组合物中各组分浓度
利用上述方法对不同浓度的A组分(含多酚的山竹提取物+含茶多酚的绿茶提取物)、B组分(含多酚的线叶金雀花提取物)单独使用时对AGEs生成的抑制率进行测定(各组分的浓度如表2所示),并计算半数抑制浓度IC50。此外,分析含多酚的山竹提取物、含茶多酚的绿茶提取物、含多酚的线叶金雀花提取物这三种组分之间的协同作用(各组分的浓度比值如表3所示)。其中,IC50add=IC50A/(P1+R×P2),式中:P1为A组分的在组合物中占的比例,P2为B组分在组合物中占的比例,P2=1-P1;R=IC50A/IC50B。将复配实验测量值IC50mix与IC50add进行比较,如果IC50mix<IC50add,则表示具有协同效应,同时计算相互作用指数(γ),用于评价协同作用程度,γ=IC50Amix/IC50A+IC50Bmix/IC50B,式中:IC50Amix、IC50Bmix分别为复配后A、B组分的IC50值,IC50A、IC50B分别为A、B组分单独作用时的IC50值。γ<1,表明A、B组分之间存在协同效应,γ值越小,表明协同作用越强。
表3抗糖化组合物中各组分的浓度和比值
注:实施例1-5中所制得的抗糖化组合物实际上与上表中的抗糖化组合物等同,即上表中的抗糖化组合物中A组分与B组分比值是由实施例1-5中抗糖化组合物中各组分的重量份数之间的比例换算而来的。
(2)抗氧化功能检测
DPPH又称1,1-二苯基-2-三硝基苯肼,是一种很稳定的以氮为中心的自由基,它的稳定性主要来自共振稳定作用的3个苯环的空间障碍,使夹在中间的氮原子上不成对的电子不能发挥其应有的电子成对作用。DPPH的无水乙醇溶液呈紫色,在517nm波长处有最大吸收峰,其吸光度与浓度呈线性关系。向其中加入自由基清除剂时,可以结合或替代DPPH,使自由基的数量减少,吸光度变小,溶液颜色变浅,借此可以评价清除自由基的能力,即通过在517nm波长处检测样品清除DPPH的效果,来计算抗氧化能力。
检测方法为:在10mL比色管中依次加入4.0mL DPPH(0.02mg/mL)的无水乙醇溶液和2.0mL实施例1-5和对比例1-6的组合物无水乙醇稀释液(100μg/mL),再加入无水乙醇至刻度,混匀立即用1cm比色皿在517nm波长处测吸光值,记为Ai,然后在室温避光保存30min后测吸光值,记为Aj,设置只加DPPH的乙醇溶液作为对照试验,其吸光值记为Ac。按照下式计算自由基清除率(K):K(%)=[1-(Ai-Aj)/Ac]*100%。自由基清除率越高,证明该组合物的抗氧化能力越强,其抗衰老效果越好。
2.实验结果
(1)非酶糖基化产物抑制结果
表4不同浓度的A组分、B组分单独使用时对AGEs的生成的抑制率
不同浓度的A组分(含多酚的山竹提取物+含茶多酚的绿茶提取物)、B组分(含多酚的线叶金雀花提取物)单独使用时对AGEs生成的抑制率结果如表4所示。由表4可知,随着浓度的增加,AGEs抑制率大体上呈现上升的趋势,A组分的半数抑制浓度IC50A=23.57μg/mL,B组分的半数抑制浓度IC50B=53.06μg/mL。
表5 AGEs抑制率和组分间的相互作用
抗糖化组合物中的含多酚的山竹提取物、含茶多酚的绿茶提取物和含多酚的线叶金雀花提取物之间的协同作用结果如表5所示。由表5可知,A组分(含多酚的山竹提取物和含茶多酚的绿茶提取物)与B组分的组合在设置的比例范围中均表现出IC50mix<IC50add,表明A组分和B组分之间存在协同效应,并且通过比较不同比例下IC50mix的大小可知,IC50值越小,表明对AGEs的抑制能力越强,抗糖化效果越强。此外,通过比较不同比例下的γ值分析协同作用,各比例下的γ值均<1,表明各比例下均存在协同效应,并且,γ值越小,表明协同效果越好。
上述结果说明本发明提供的抗糖化组合物能够有效抑制AGEs的生成,并且其中的含茶多酚的绿茶提取物、含多酚的山竹提取物、含多酚的线叶金雀花提取物之间具有良好的协同抗糖化作用。
(2)抗氧化功能
由表6的自由基清除率可知,同时含有含多酚的山竹提取物、含茶多酚的绿茶提取物和含多酚的线叶金雀花提取物的抗糖化组合物的自由基清除率在90%以上,并且实施例1-5所得到的组合物的自由基清除率显著高于对比例1-6所得到的组合物。由此可知,本发明提供的抗糖化组合物能够有效地清除自由基,具有较强的抗氧化作用,抗衰老效果好。
表6自由基清除率
样品 | K(%) | 样品 | K(%) |
实施例1 | 91.2 | 对比例1 | 72.1 |
实施例2 | 92.3 | 对比例2 | 73.5 |
实施例3 | 95.7 | 对比例3 | 74.3 |
实施例4 | 93.4 | 对比例4 | 81.6 |
实施例5 | 93.9 | 对比例5 | 83.5 |
对比例6 | 85.1 |
以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照上述实施例对本发明进行了详细的说明,所属领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,但这些修改或替换均在本发明的保护范围之内。
Claims (10)
1.一种抗糖化组合物,其特征在于,包括以下重量份的组分:含多酚的山竹提取物0.5-5份、含茶多酚的绿茶提取物0.5-5份、含多酚的线叶金雀花提取物1份。
2.如权利要求1所述抗糖化组合物,其特征在于,包括以下重量份的组分:含多酚的山竹提取物1-2.5份、含茶多酚的绿茶提取物1-2.5份、含多酚的线叶金雀花提取物1份。
3.如权利要求1所述抗糖化组合物,其特征在于:所述含多酚的山竹提取物中多酚含量≥0.01%,所述含茶多酚的绿茶提取物中茶多酚含量≥50%,所述含多酚的线叶金雀花提取物中多酚含量≥15%。
4.如权利要求1所述抗糖化组合物,其特征在于:所述含多酚的山竹提取物由以下步骤制备得到:将山竹加入溶剂进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到所述含多酚的山竹提取物;
所述含茶多酚的绿茶提取物由以下步骤制备得到:将绿茶加入溶剂进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到所述含茶多酚的绿茶提取物;
所述含多酚的线叶金雀花提取物由以下步骤制备得到:将线叶金雀花加入溶剂进行提取,得到混合液,过滤,弃去滤渣,浓缩,干燥,得到所述含多酚的线叶金雀花提取物。
5.如权利要求1-4任意一项所述抗糖化组合物的制备方法,其特征在于,包括以下步骤:将所述含多酚的山竹提取物、所述含茶多酚的绿茶提取物、所述含多酚的线叶金雀花提取物混合均匀,得到所述抗糖化组合物。
6.如权利要求1-4任意一项所述抗糖化组合物在制备抗糖化食品、抗糖化保健品中的应用。
7.一种抗糖化食品,其特征在于:包括如权利要求1-4任意一项所述抗糖化组合物,所述抗糖化组合物在所述抗糖化食品中的重量占比为0.01-25%。
8.如权利要求7所述抗糖化食品,其特征在于:所述抗糖化食品包括饮品、果冻、固体饮料、压片糖果、软糖。
9.一种抗糖化保健品,其特征在于:包括如权利要求1-4任意一项所述抗糖化组合物,所述抗糖化组合物在所述抗糖化保健品中的重量占比为0.01-25%。
10.如权利要求9所述抗糖化保健品,其特征在于:所述抗糖化保健品包括口服液、胶囊、片剂、粉剂。
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