CN110051001A - 一种用于改善记忆的膳食补充剂 - Google Patents
一种用于改善记忆的膳食补充剂 Download PDFInfo
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- CN110051001A CN110051001A CN201910343731.9A CN201910343731A CN110051001A CN 110051001 A CN110051001 A CN 110051001A CN 201910343731 A CN201910343731 A CN 201910343731A CN 110051001 A CN110051001 A CN 110051001A
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Abstract
一种用于改善记忆的膳食补充剂，其各成分及重量百分含量为：神经肽30‑50％、微量元素及维生素2‑5％、不饱和脂肪酸10‑20％和助剂25‑60％；其中，所述肽配制品中包含以下几种序列的肽：EVGEIE、QKQLQ、LERNAIKKVGK和TEPGLE，本发明的膳食补充剂易于吸收、无明显副作用，可以保持神经细胞的活性，使其免受衰老相关损伤；用于预防、改善或对抗与衰老过程相关的缺陷，激活与记忆力相关的大脑机制，改善长期记忆力和程序性记忆力以及学习表现、注意力，在衰老过程中保持健康的精神。
Description
技术领域
本发明属于膳食补充剂领域，具体涉及一种用于改善记忆的膳食补充剂。
背景技术
随着经济发展，人民的整体生活水平大大提高，人民的寿命普遍延长，同时加快了我国社会老龄化的发展速度。
我国早在1999年就已经步入老龄化社会，老龄人口逐年增加。2010年第六次全国人口普查结果显示，全国大于等于60岁的老年人口达1.78亿，占总人口数的13.26％。2050年，我国60岁以上老年人将达4.51亿。全球范围老龄人口的增加导致老年痴呆患者也随之增加，美国开展了2050年阿尔茨海默症患病人数的预测研究，2050年美国65岁以上老年人患阿尔茨海默病人数达到1380万，是2010年患病人数的3倍，可见患病人数增长迅速。
随着社会老龄化的发展，各种慢性躯体性疾病影响着老年人的生命健康的同时，精神卫生问题也成为老年人生命健康的一大威胁，其中，痴呆症在老年人精神疾患中最为普遍和严重。老年痴呆是继肿瘤、心脏病、脑血管疾病之后引起老年人死亡的第四大病因。它可以引起老年人发生持续时间较长的智能损害，表现为记忆、语言、思维、计算、定向力和情感障碍及人格改变，并出现社会活动能力和自身生活能力的减退。伴随病情加剧，病人变得呆滞、无欲、长时间卧床、生活无法自理，常因褥疮、骨折、肺炎、泌尿系统感染、营养不良等继发性躯体疾病或因衰竭而死亡。
老年痴呆患者的增加将导致疾病经济负担增加，美国国家脑研究基金会估计，美国每年用于痴呆的花费超过1130亿美元，直接花费超180亿美元，间接费用估计超过940亿美元。据预测，2050年我国老年痴呆的直接经济负担每年将达7453.21亿元。老年痴呆带来的沉重经济负担和社会负担，引起了科学家的关注和研究，成为全世界关注的社会、卫生、经济问题。
目前对于老年痴呆的治疗药物主要包括ACh EI、谷氨酸受体调节剂、钙拮抗剂、金属螯合剂、改善脑循环和脑代谢药物、疫苗和基因治疗药物等，其中还有其他如中医治疗药物、精神抑制剂、降胆固醇药、抗氧化剂等。
但是，观察这些药物治疗AD的作用效果，发现没有任何一种药物能够有效治疗，它们都只是在一定程度上缓解痴呆症状，延缓疾病进展。2018年初，辉瑞制药公司也宣布关于阿尔茨海默症的潜伏期、I期、II期的早期临床试验全部失败。
因此，迫切需要一种新形式的、有效的、非药物的补充剂供老年人食用。
发明内容
本发明的目的在于提供一种用于改善记忆的膳食补充剂，其易吸收、无明显副作用，可以保持神经细胞的活性，使其免受衰老相关损伤；用于预防、改善、对抗与衰老过程相关的缺陷，激活与记忆力相关的大脑机制，预防、对抗及改善认知功能下降，支持、维持和改善长期记忆力和程序性记忆力以及学习表现、注意力，在衰老过程中保持健康的精神。
为了达到上述目的，本发明提供给以下技术方案：
一种用于改善记忆的膳食补充剂，其各成分及重量百分含量为：神经肽30-50％、微量元素及维生素2-5％、不饱和脂肪酸10-20％和助剂25-60％；其中，所述神经肽中包含以下几种序列的肽：EVGEIE、QKQLQ、LERNAIKKVGK和TEPGLE，四者的重量比例为：EVGEIE：QKQLQ：LERNAIKKVGK：TEPGLE＝1-3：1-3：1-2：1-3。
进一步，所述不饱和脂肪酸为二十二碳六烯酸、亚油酸、亚麻酸、二十碳五烯酸(EPA)。
又，所述维生素选自维生素A、维生素B、维生素C、维生素D和维生素E中的一种或几种；所述的微量元素选自碘、锌、硒、铜、钼、铬、钴、铁、锰、硅、硼、钒和镍中的一种或几种。
优选地，所述助剂为防腐剂、调味剂、着色剂、添加剂、填充剂和赋形剂中的一种或几种。
进一步，所述防腐剂选自苯甲酸、苯甲酸钠、山梨酸、山梨酸钾或对羟基苯甲酸乙酯；调味剂选自柠檬酸或苹果酸；着色剂选自胭脂红或柠檬黄；添加剂选自丁基羟基茴香醚或二丁羟基甲苯；填充剂选自淀粉或琼脂；赋形剂选自卡拉胶或明胶。
本发明所述膳食补充剂的剂型为片剂、包衣片剂、胶囊、糊剂、咀嚼片或饮用溶液。
优选地，所述包衣片剂为肠包衣剂型。
线粒体应激反应强弱与衰老和记忆状态直接相关，而痛敏肽通过与痛敏肽受体结合可减弱线粒体应激反应，加速衰老及记忆减退，本发明的膳食补充剂中，添加的神经肽制品可以抑制痛敏肽及痛敏肽受体的结合，进而增强线粒体的应激反应，延缓衰老，改善记忆；添加的活性肽易吸收、功能强、无明显副作用，非常适合作为老年人的膳食补充剂。
本发明膳食补充剂中，包含了平衡量的具有神经保护活性的不饱和脂肪酸、微量元素及维生素；选择的二十二碳六烯酸(DHA)具有辅助脑细胞发育、抗衰老、改善血液循环、降血脂的作用，是人体所必需的多不饱和脂肪酸。
与现有技术相比，本发明具有如下有益效果：
本发明的膳食补充剂具有神经保护作用，其中的神经肽制品可减少神经细胞衰老，保持神经细胞活性，使其免受衰老相关损伤，预防、改善、对抗与衰老过程相关的缺陷；激活与记忆力相关的大脑机制，预防、对抗和/或改善认知功能下降。
本发明的膳食补充剂能够支持、维持和/或改善长期记忆力和程序性记忆力以及学习表现、注意力，并且用于在衰老过程中保持或支持健康的精神功能，改善老年小鼠的记忆功能，适于改善老年人记忆。
附图说明
图1为本发明实施例1中光镜下观察的神经球。
图2为本发明实施例1中不同剂量本发明膳食补充剂对27-羟基胆固醇(27HC)诱导神经细胞衰老的保护作用。
图3为本发明实施例6中两组不同训练日逃逸潜伏期比较。
图4为本发明实施例6中两组小鼠穿越第三象限和平台的次数比较。
图5为本发明实施例6中两组小鼠停留在第三象限和平台的时间比较。
具体实施方式
以下结合具体实施例对本发明作进一步说明。
购买无菌溶液，并且在层流橱中混合终溶液；在试验前对所有必需物品进行灭菌。
用于大批分析的培养基由DMEM(Dulbecco's Modified Eagle Medium)培养基与10g/L的B27和N2、0.01％庆大霉素组成，其中加入庆大霉素以防止培养基被支原体或其它有害微生物污染，每次试验都在无菌条件下的层流橱中制备新鲜的营养培养基。
实施例1一种改善记忆的膳食补充剂
本实施例的膳食补充剂的具体配方：
神经肽40％、微量元素及维生素5％、不饱和脂肪酸20％和助剂35％，其中，所述的神经肽由比例为1：1：1：1的EVGEIE、QKQLQ、LERNAIKKVGK和TEPGLE组成。
用于试验的细胞是小鼠脑皮层神经干细胞，取小鼠胚胎14-15天的胎鼠脑皮层组织，用无菌手术器械将其剪成黄豆大小，并用200目不锈钢筛网过滤后种植于培养瓶中，37℃，5％CO2培养，5-7天待原代克隆形成后，采用机械分离方法传代，经吸管吹打，200目不锈钢筛网过滤，800r/min离心10min，弃上清，加入新鲜的神经球培养基，用吸管反复小心吹打制成单细胞悬液，以1：2比例传代(图1)。
27HC能通过血脑屏障进入脑内，高浓度的27HC能够损害神经细胞，促进神经细胞衰老继而导致生物体的衰老，10μM27HC可导致约60％的神经细胞衰老。
将初始组织培养基从细胞中除去，实验组加入含有10μM27HC的新培养基，并与细胞一起培养12小时，对照组分别加入不同浓度的本实施例膳食补充剂(浓度分别为0.5mg/ml、1mg/ml、2mg/ml、4mg/ml)与含有10μM 27HC的培养基共同培养12小时，此后观察每组的衰老阳性细胞数，结果参见图2。
由图2可见，与不同浓度的本发明膳食补充剂培养12小时后，衰老阳性细胞数比例明显下降，在4mg的高剂量组中衰老阳性细胞约10％，这表明多数的神经细胞可以被挽救，所有的剂量都具有明显的神经保护作用，说明本发明的膳食补充剂可以保持神经细胞活性，保护它们免受衰老相关损伤。
实施例2一种改善记忆的膳食补充剂片剂的制备方法
本实施例的膳食补充剂包括：神经肽50％、微量元素及维生素5％、不饱和脂肪酸15％和助剂30％，所述的神经肽由比例为2：2：1：2的EVGEIE、QKQLQ、LERNAIKKVGK和TEPGLE组成。
用1.2mm筛网将上述混合物粉末50kg，筛选到不锈钢筒中，并用翻滚型混合器以6rpm的混合速度混合10分钟，用1.2nm筛网将1kg硬脂酸镁筛选到上述混合物的不锈钢筒中。随后，用翻滚型混合器以4rpm的混合速度将所有的颗粒混合5分钟得到膳食补充剂混合物。
用约6万片/小时的压片速度压片机压制得到的混合物，使用圆形、双凸面冲头，其直径为11mm。
片剂的特性为(所有性能测试方法按照中国药典进行)：
实施例3片剂包衣的制备：
对实施例2中的片剂制作包衣。
将以下成分用推进式混合器进行混合以形成薄膜包衣悬浮液：
17.5kg 醋钛纤维素(30％水性溶液)；
0.44kg 滑石粉；
0.621kg 二氧化钛；
1.059kg 柠檬酸三乙酯。
在转鼓式包衣机中用薄膜包衣悬浮液包衣片剂至终重量为420mg/膜片。包衣片可用蜂蜡或石蜡打光。以这种方式，得到具有以下参数的薄膜包衣的白色片剂(所有性能测试方法按照中国药典进行)：
平均重量： 420±15mg；
崩解： ＜30min；
脆性： ＜1％。
实施例4胶囊制剂的制备
将实施例2的粉末混合物装填到硬明胶胶囊中，用385mg的粉末混合物装填0号或1号硬明胶胶囊。从而获得具有以下参数的胶囊(所有性能测试方法按照中国药典进行)：
平均重量：460±28.8mg(1号)；
平均重量：480±28.8mg(0号)；
崩解：＜15min；
或者，相当量的颗粒可使用2号和3号硬明胶胶囊。
实施例5饮用溶液的制备
将含有神经肽30％、微量元素及维生素5％、不饱和脂肪酸10％和助剂55％，所述的神经肽由比例为3：3：1：3的EVGEIE、QKQLQ、LERNAIKKVGK和TEPGLE组成的混合物500g溶于盛有1500g水的玻璃烧瓶中，该烧瓶装有推进式混合器,形成几乎澄清的溶液，并加水稀释至5000ml，将其通过0.45μm的薄膜滤器进行过滤以进一步澄清。将该溶液装填于带有螺旋瓶盖的20ml玻璃瓶中制成引用溶液。
实施例6本发明膳食补充剂用于老年小鼠
1.实验方法
实验对象为18(±1)月龄的C57BL/6小鼠，将小鼠随机分配至实验组和对照组中，每组15只，实验组每天用本发明实施例1的膳食补充剂0.2mg灌胃1次，对照组每天用生理盐水灌胃1次，持续3个月，之后进行水迷宫实验、定位航行实验、空间探索实验。
1.1水迷宫实验
进行水迷宫实验时，应保持周边环境安静、池内水温适宜，并且于水迷宫所在之房间内的水池周围贴上特殊符号的贴纸作为空间参照的线索。
所采用的水迷宫装置为不锈钢圆形水池，其直径为116cm，高为60cm，人工分为四个象限，并于第三象限的中点、水面下0.5-1cm的位置放置一个直径10cm的圆形站台，水池在一个光亮的屋内，在池内加入白色无毒染料使水不透明。
1.2定位航行实验
前5天进行定位航行实验，每天于固定的时间随机从4个象限将小鼠放入水中，观察其游泳和找寻目标平台的情况，共1分钟，记录小鼠从入水到找到平台的时长，此为逃避潜伏期。若小鼠在进入水池后超过2分钟仍未能顺利找到圆形站台并登台，则将其引导到圆形站台休息30秒，使其熟悉隐蔽站台的位置；潜伏期记为最高分120秒，记录小鼠寻找固定平台所需时间和速度。每天训练1次，持续5天。记录每次训练的游泳速度和逃避潜伏期。以视频采集仪器记录并分析实验小鼠的逃避潜伏期和游泳路径。
1.3空间探索实验
定位航行实验进行完毕后，开始空间探索实验，撤除第三象限的圆形站台后，将所有实验小鼠由四个象限分别放入圆形水池内，观察并记录其1分钟内于水池中的全程游泳路径。以应用软件分析其在隐蔽站台所在象限(第三象限)的停留时间，以及穿过隐蔽站台位置的穿台次数等相关参数，进行相关统计学分析。
2.结果
2.1不同处理方式对老年小鼠逃逸潜伏期及游泳速度的影响
水迷宫定向航行实验显示，随着训练次数的增加，两组小鼠寻找平台的潜伏期时间均呈现明显缩短趋势，以本发明膳食补充剂灌胃组缩短较为明显(参见图3)。
此外，实验还测量了两组小鼠的游泳速度，发现两组小鼠的游泳速度没有明显差异，因此，不会影响两组的逃逸潜伏期。
训练就是让小鼠学习并且记住平台位置的过程，从结果来看，随着训练次数的增加，两组小鼠寻找平台的潜伏期都缩短，这说明两组小鼠都在学习后一定程度上记住了平台的位置。
另外，经过对比，以本发明膳食补充剂灌胃组小鼠寻找平台潜伏期时间缩短的更多，也就是本发明膳食补充剂灌胃后小鼠学习记忆能力更强，所以本发明膳食补充剂能够改善学习记忆。
2.2不同处理方式对老年小鼠穿越平台次数及停留时间的影响
在水迷宫空间探索实验中，与对照组比较，本发明膳食补充剂灌胃实验组小鼠穿越第三象限和原平台次数分别增加了100％和200％(图4)，在第三象限和原平台位置停留时间分别增加了87％和120％(图5)。
也就是说，在定速巡航试验的时候平台是放在第三象限的，而经过5天的训练后撤去了平台进行空间探索试验，这时候小鼠并不知道平台已经撤走，它习惯性的依靠空间记忆去寻找第三象限里的平台，所以穿越第三象限和原平台次数多、在第三象限和原平台位置停留时间长的就是记忆力好的小鼠，通过结果可知，正是本发明膳食补充剂灌胃组小鼠记忆力较对照组小鼠记忆力好，说明本发明膳食补充剂能够改善老年小鼠的记忆功能。
Claims (7)
1.一种用于改善记忆的膳食补充剂，其各成分及重量百分含量为：神经肽30-50％、微量元素及维生素2-5％、不饱和脂肪酸10-20％和助剂25-60％；其中，所述神经肽中包含以下几种序列的肽：EVGEIE、QKQLQ、LERNAIKKVGK和TEPGLE，四者的重量比例为：EVGEIE：QKQLQ：LERNAIKKVGK：TEPGLE＝1-3：1-3：1-2：1-3。
2.根据权利要求1所述用于改善记忆的膳食补充剂，其特征在于，所述不饱和脂肪酸为二十二碳六烯酸、亚油酸、亚麻酸或二十碳五烯酸。
3.根据权利要求1所述用于改善记忆的膳食补充剂，其特征在于，所述维生素选自维生素A、维生素B、维生素C、维生素D和维生素E中的一种或几种；所述微量元素选自碘、锌、硒、铜、钼、铬、钴、铁、锰、硅、硼、钒和镍中的一种或几种。
4.根据权利要求1所述用于改善记忆的膳食补充剂，其特征在于，所述助剂为防腐剂、调味剂、着色剂、添加剂、填充剂和赋形剂中的一种或几种。
5.根据权利要求1所述用于改善记忆的膳食补充剂，其特征在于，所述防腐剂选自苯甲酸、苯甲酸钠、山梨酸、山梨酸钾或对羟基苯甲酸乙酯；调味剂选自柠檬酸或苹果酸；着色剂选自胭脂红或柠檬黄；添加剂选自丁基羟基茴香醚或二丁羟基甲苯；填充剂选自淀粉或琼脂；赋形剂选自卡拉胶或明胶。
6.根据权利要求1所述用于改善记忆的膳食补充剂，其特征在于，所述膳食补充剂的剂型为片剂、包衣片剂、胶囊、糊剂、咀嚼片或饮用溶液。
7.根据权利要求6所述用于改善记忆的膳食补充剂，其特征在于，所述包衣片剂为肠包衣剂型。
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