CN111920878B - 一种抗衰老组合物及其应用 - Google Patents
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Abstract
本发明涉及医药学技术领域,尤其涉及一种抗衰老组合物及其应用。该抗衰老组合物的活性成分为烟酰胺单核苷酸和葡萄籽提取物的复配产物。该抗衰老组合物可以进一步提高烟酰胺单核苷酸的利用效率及转化效率,提高体内NAMPT含量,上调NAD+含量,具有抗衰老的优异效果;同时降低了现有抗衰老补充剂的成本。
Description
技术领域
本发明涉及医药学技术领域,尤其涉及一种抗衰老组合物及其应用。
背景技术
衰老是一种渐进的过程,直到最近才被认为伴随着不可避免的退行性改变,成为重要的研究领域之一。基于全球人口的统计数据表明,1900年出生的人的平均寿命是47岁,从那时起,科学和医学方面的改进和发现每十年可以使人类平均寿命增加2年左右,这也就是说现在出生的孩子预计可以活到100岁,这对人类社会及经济都有巨大影响,而对生物及医学的研究提出了新的要求[1]。安全且有效地干扰衰老进程的有害影响的治疗策略引起了广泛的关注,有一些潜在有效的研究项目,例如选择性消除衰老细胞或抑制衰老相关蛋白功能,已在临床前期研究中证明具有良好的治疗效果[2,3]。
衰老作为包括肿瘤、心血管疾病、糖尿病及神经退行性改变在内的诸多疾病的发生及发展中发挥中重要作用,也吸引着越来越多的科研人员的注意力。然而,衰老的进程是一个复杂的过程,什么驱动衰老过程?遗传学、端粒磨损或者有毒产物过载?不同的理论提出了不同的思路。尽管衰老的研究尚缺乏足够的共识,目前的研究结果提出,科研将九种衰老的细胞和分子表型认为是年龄相关功能障碍的根本原因,可以用于协助衰老相关的科学研究设计及潜在的治疗干预措施。这些标志包括:基因组不稳定性、端粒磨损、表观遗传改变、蛋白质稳态丧失、营养物质感知失调、线粒体功能障碍、细胞衰老、干细胞衰竭和细胞间通讯改变[4]。NAD+又叫辅酶Ⅰ,全称烟酰胺腺嘌呤二核苷酸,是细胞内DNA修复系统的重要原料,也是细胞核与线粒体间的关键联络因子,作为抗衰老研究领域最受关注的靶点得到广泛研究[5]。然而,NAD+作为大分子难以直接通过细胞膜被利用,因此,科学家们将目光投向了NAD+的前体物质β-烟酰胺单核苷酸(NMN)。NMN作为NAD+的前体,其功能也是通过NAD+来体现。近年来基于人体和动物表明,外源性补充NMN可有效地恢复体内辅酶I水平,延缓衰老并预防多种退行性病变[6]。此外,进一步的研究也证实了其在恶性肿瘤、肥胖、心血管损伤及糖尿病等疾病中具有潜在的保护作用[6]。然而,NMN在体内调节复杂,如何进一步提高其利用效率仍是目前研究的重点。
NAMPT又称为内脏脂肪素(visfatin)和前B细胞克隆增强因子(pre-B cellenhancing factor,PBEF),是NAD+补救合成途径中催化NMN合成为NAD+的限速酶,NAMPT含量下降在衰老进程中处于关键作用[7]。激发NAMPT活性可显著抑制动脉粥样硬化、骨骼肌衰老及神经退行性改变病程,是抗衰老领域极具潜力的方向[8]。
葡萄籽提取物(GSE)是一种天然植物化学物质,目前已被美国食品和药物管理局(FDA)公认为安全的化学物质[9]。葡萄籽是原花青素,主要由单体儿茶素和表儿茶素、没食子酸、聚合和低聚原花青素组成,这些原花青素被证明比维生素C、E和β-胡萝卜素更强大的自由基清除剂[10]。GSE对癌症和代谢细胞的影响已被大量研究,例如在哺乳动物中,GSE在体内对氧化应激和许多代谢紊乱(包括胰岛素抵抗)产生有益影响,这些代谢紊乱可能与调节血浆脂联素(如脂联素)有关[11,12]。此外,GSE也显著抑制产气荚膜菌活性进而广泛地应用于食品的防腐中[13]。最近的一项基于小鼠的动物实验表明,饮食中添加GSE可以提高血浆中NAMPT含量,然而对NAD+含量是否产生影响尚不清楚。
参考文献
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发明内容
本发明提供了一种抗衰老组合物及其应用,该抗衰老组合物可以进一步提高烟酰胺单核苷酸的利用效率,提高体内NAMPT含量,上调NAD+含量,具有抗衰老的优异效果;同时降低了现有抗衰老补充剂的成本,解决了现有技术中存在的问题。
本发明所采用的技术方案之一是:
一种抗衰老组合物,其活性成分包括以下重量份数的原料:
烟酰胺单核苷酸1份、葡萄籽提取物0.5-2份。
进一步的,所述的抗衰老组合物,其活性成分包括以下重量份数的原料:烟酰胺单核苷酸1份、葡萄籽提取物2份。
进一步的,所述的抗衰老组合物的活性成分包括以下重量份数的原料:烟酰胺单核苷酸1份、葡萄籽提取物2份。
进一步的,所述葡萄籽提取物为原花青素质量含量85%-100%的葡萄籽提取物。
进一步的,所述的抗衰老组合物,还包括填料,所述填料包括稳定剂和调味剂。
进一步的,所述填料包括维生素C、蓝莓、木糖醇、羟丙基纤维素、聚维酮、硬脂酸镁、二氧化钛的一种或多种。
进一步的,上述的抗衰老组合物的制备方法,采用将各活性成分混合、搅拌均匀得到。
本发明所采用的技术方案之二是:
上述抗衰老组合物在制备抗衰老保健品或抗衰老药品方面的应用。
进一步的,所述抗衰老组合物的服用剂量不低于300mg/Kg体重,优选服用剂量600mg/Kg体重。
进一步的,所述抗衰老组合物还包括用于改善制剂稳定性和口感的填料。
上述的抗衰老组合物在制备提升体内NAMPT、上调NAD+含量的保健品或药品方面的应用。
本发明的有益效果:
本发明的抗衰老组合物采用烟酰胺单核苷酸和葡萄籽提取物以特定比例的复配获得,可以进一步提升烟酰胺单核苷酸的利用效率,促进烟酰胺单核苷酸转化为NAD+,同时提高体内NAMPT含量,上调NAD+含量,具有显著的抗衰老的优异效果,而且降低了现有抗衰老补充剂的成本。
本发明以NMN及NAMPT活力为切入点,关注NMN转化为NAD+的关键反应,选取了相对成熟的天然提取物葡萄籽提取物,与NMN配合具有显著的协同增效作用,有望通过同时提高反应底物及酶活性的方式来降低消费者费用,并维持稳定且高效的抗衰老效果。
附图说明
图1为在衰老小鼠模型上使用GSE对大脑、肝脏及肌肉中NAMPT含量的影响;*:P<0.05;
图2为在衰老小鼠模型上使用NMN+GSE对大脑、肝脏及肌肉中NAD+含量的影响;*:P<0.05。
具体实施方式
为能清楚说明本方案的技术特点,下面通过具体实施方式,结合附图,对本发明进行详细阐述。
实施例1动物实验
1.材料:
实验动物:选择不同年龄的清洁级C57BL/6小鼠,5只1月龄,体重(21±2)g;20只16月龄,体重(38±5)g,购自上海斯莱克实验动物有限责任公司,饲养于上海同济大学动物中心清洁级环境,适应性饲养2周后进入实验。实验中对于实验动物的使用和操作均遵守国家技术委员会颁布的《实验动物管理条例》并符合实验动物护理与使用规范。
2.方法:
2.1动物分组及干预:按照开始实验时小鼠年龄,将所有小鼠分为以下几组:年轻小鼠(6周龄,5只)、老龄小鼠(18周龄,5只)、NMN干预老龄小鼠组(18周龄,5只):200mg/Kg体重、GSE干预老龄小鼠(18周龄,5只),GSE干预组每日口服GSE50 mg/kg体重、GSE100mg/kg体重、GSE200mg/kg体重或者GSE400mg/kg体重,干预时间2周,NMN+GSE干预老龄小鼠(18周龄,5只)NMN及GSE含量分别为:200mg/Kg NMN+50mg/KgGSE/、200mg/Kg NMN+100mg/Kg GSE、200mg/Kg NMN+200mg/Kg GSE及200mg/Kg NMN+400mg/Kg GSE。
2.2组织获取及蛋白提取:各组小鼠按10%水合氯醛3mL/kg腹腔麻醉,固定四肢和头部,剪开胸骨充分暴露心脏,立即用生理盐水心脏灌注,同时剪开右心耳放血,心脏冷生理盐水灌流待后迅速获取小鼠大脑、肝脏及肌肉组织。组织称重,切小块放入管中,配置含抑制剂的蛋白质抽提试剂(1mL抽提试剂中加入5μL蛋白酶抑制剂混合液,5μLPMSF和5ul磷酸酶混合液)。加入预冷的含抑制剂的蛋白质抽提试剂(250mg组织中加入1ml抽提试剂)。用匀浆器每次30秒低速匀浆,每次匀浆间隔冰浴1分钟,至组织完全裂解。裂解液于预冷的离心机中14000g离心15分钟。上清液立刻转移入新的离心管中保存待用。
2.3NAMPT含量检测:NAMPT含量通过ELISA试剂盒(RayBio Human Visfatin EIA–VIS–1,Norcross,GA,USA)进行检测,具体操作参考试剂盒说明书。
2.4NAD+含量:NAD+含量检测通过检测试剂盒完成(Abcam;Cambridge,MA,USA;catalog#:Ab65348),具体参照参考试剂盒说明书,按照如下公式进行NAD+含量计算:[NAD+]=[NADtotal]-[NADH]。
2.5统计方法:数据以均数±标准误(mean±SEM)表示,采用SPSS 20.0for Window软件作单因素方差分析(one-way ANOVA)检验差异显著性,多个样本均数之问比较采用Student-Newman-Keuls(SNK)检验,P<0.05为有统计学意义。
3.结果
3.1NMN+GSE显著提高衰老小鼠中NAMPT含量
与6周龄的年轻小鼠相比,18周龄小鼠的大脑、肝脏及肌肉中的NAMPT含量显著下降,分别为年轻小鼠的0.23±0.09、0.25±0.09及0.21±0.1倍(P<0.05)。
单独补充NMN对NAMPT含量无显著影响,而18周龄对照小鼠相比,口服50mg/Kg GSE对各组织中NAMPT含量无显著影响,而100mg/Kg GSE、200mg/Kg及GSE400mg/Kg GSE显著提高了大脑、肝脏及肌肉NAMPT含量(100mg/Kg GSE组与年轻小鼠的比值分别为0.68±0.14、0.73±0.19及0.76±0.12,P<0.05;200mg/Kg GSE组与年轻小鼠的比值分别为0.89±0.13、0.85±0.16及0.87±0.17,P<0.05;400mg/Kg GSE组与年轻小鼠的比值分别为0.89±0.17、0.9±0.16及0.88±0.16,P<0.05)(图1)。
3.2NMN+GSE显著提高衰老小鼠中NAD+含量
如图2所示,与6周龄的年轻小鼠相比,18周龄小鼠的大脑、肝脏及肌肉中的NAD+含量显著下降,分别为年轻小0.23±0.09、0.25±0.09及0.21±0.1的倍(P<0.05)。
单独补充GSE对NAD+含量无显著影响,而18周龄对照小鼠相比,口服NMN显著提高了大脑、肝脏及肌肉NAD+含量(各组与年轻小鼠的比值分别为0.56±0.11、0.55±0.13及0.62±0.19,P<0.05)。
进一步的研究显示,与单纯NMN 200mg/kg干预组相比,NMN 200mg/kg+GSE50mg/kg组合未能显著提高各组织中NAD+含量,而NMN 200mg/kg+GSE 100-400mg/kg组合中均能显著提高各组织的NAD+含量,与NMN与GSE配比2:1至于1:2各组合取得了相近的保护效果(P>0.05)。
4.结论:
NMN+GSE显著提高衰老小鼠中NAMPT及NAD+含量。
综上:本发明通过体体内实验的证据证明,NMN+GSE特定比例的联合使用可以同时显著提高NAMPT以及NAD+含量。两种关键的活性成分均是已上市的成分,故有相当的安全性。而与传统的抗氧化剂为主的抗衰老药物相比,添加的NMN显著的靶向NAD+形成这一关键通路进而发挥调控左右。相对于单纯NMN补充剂,本产品含有的GSE成分可以显著提高NAMPT含量,进而促进NMN发挥保护作用,在控制费用的情况下具有潜在的更强的抗衰老效果。
上述具体实施方式不能作为对本发明保护范围的限制,对于本技术领域的技术人员来说,对本发明实施方式所做出的任何替代改进或变换均落在本发明的保护范围内。
本发明未详述之处,均为本技术领域技术人员的公知技术。
Claims (8)
1.一种抗衰老组合物,其特征在于,其活性成分由以下重量份数的原料制成:
烟酰胺单核苷酸1份、葡萄籽提取物0.5-2份;
所述葡萄籽提取物为原花青素质量含量85%~100%的葡萄籽提取物。
2.根据权利要求1所述的抗衰老组合物,其特征在于,其活性成分由以下重量份数的原料制成:烟酰胺单核苷酸1份、葡萄籽提取物2份。
3.根据权利要求1或2所述的抗衰老组合物,其特征在于,还包括填料,所述填料包括稳定剂和调味剂。
4.根据权利要求3所述的抗衰老组合物,其特征在于,所述填料包括维生素C、蓝莓、木糖醇、羟丙基纤维素、聚维酮、硬脂酸镁、二氧化钛的一种或多种。
5.如权利要求1或2所述的抗衰老组合物在制备抗衰老保健品或抗衰老药品方面的应用。
6.根据权利要求5所述的应用,其特征在于,所述抗衰老组合物的服用剂量不低于300mg/Kg体重。
7.根据权利要求5所述的应用,其特征在于,所述抗衰老组合物还包括用于改善制剂稳定性和口感的填料。
8.如权利要求1或2所述的抗衰老组合物在制备提升体内NAMPT、上调NAD+含量的保健品或药品方面的应用。
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