CN108471795A - 含有葡萄籽提取物、缬草提取物和红花籽提取物的用于预防和缓解女性绝经期症状的食品组合物 - Google Patents
含有葡萄籽提取物、缬草提取物和红花籽提取物的用于预防和缓解女性绝经期症状的食品组合物 Download PDFInfo
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- Non-Alcoholic Beverages (AREA)
Abstract
本发明涉及含有葡萄籽提取物、缬草提取物和红花籽提取物的用于预防和缓解女性绝经症状的食品组合物。本发明的含有葡萄籽提取物、缬草提取物和红花籽提取物的复合材料具有抑制破骨细胞分化、促进成骨细胞活性、增加雌激素受体表达以及增加IGF‑1和骨钙蛋白表达的优异效果,从而表现出预防和缓解女性绝经期症状的效果。另外,本发明可提供具有预防和缓解女性绝经期症状的优异效果并含有复合材料作为活性成分的食品组合物,所述复合材料含有无毒、无副作用且对人体无害的天然提取物。
Description
技术领域
本发明涉及含有葡萄籽提取物、缬草提取物和红花籽提取物的用于预防和缓解女性绝经期症状的食品组合物。
背景技术
随着女性年龄增长,卵巢功能劣化,从而导致雌激素分泌大幅减少。因此,女性对促卵泡激素(FSH)和促黄体生成激素(LH)的响应以及黄体酮(progesterone)产生降低。特别是,在她们50岁左右,女性经历绝经期的可能性很高,这是卵巢功能完全停止的时间。从绝经开始,绝经后激素平衡被破坏,导致各种症状。这被称为“更年期(climacterium)”。此外,由卵巢切除术和子宫切除术等引起的早期绝经也可能导致更年期症状。
也就是说,女性绝经期是一种内分泌综合征,意味着作为女性激素的雌激素由于卵巢功能老化而下降,并且生理和性功能减退或丧失。该更年期症状包括由血管变化引起的症状如面部潮红、心动过速、出汗或头痛,由肌肉骨骼变化引起的症状如肌痛、关节痛和背痛,由泌尿生殖器变化引起的症状如尿频或尿失禁,以及由于脑-神经系统变化引起的症状如导致记忆减退、抑郁症、注意力丧失和头晕。另外,出现诸如视力丧失以及皮肤和毛发变化的症状,并且出现由激素变化引起的骨质疏松症和对女性健康致命的疾病如心血管疾病。
近期的文明发展和科学进步导致了现代人类的预期寿命延长。由于2002年女性的平均预期寿命为80.4岁,40多岁后的30-40年间逐渐开始出现绝经期症状,因此需要特别注意女性的健康生活。
由于雌激素缺乏症是绝经期症状的主要原因,因此雌激素替代疗法主要用作改善女性绝经期症状的补救措施。然而,仅有35%至40%的具有绝经期症状的女性使用雌激素替代疗法,乃至她们不愿意继续使用这种方法(Ann.Intern.Med.1999.130:545-553)。雌激素替代疗法基于人工激素的施用,因此导致排斥响应和副作用。据报道,许多研究表明雌激素替代疗法的不良反应可能增加子宫出血、中风、心脏病发作、乳腺癌和子宫癌的风险(Obstet.Gynecol.1998.91:678-684,New Engl.J.Med.1995.332:1589-1593)。
由于这些问题,人们越来越希望用自然摄入食品的方法来代替雌激素疗法,所以需要开发用于治疗绝经期症状的药剂,这对于减轻更年期症状且不引起副作用是非常有效的,因此大量的研究正在寻找天然植物成分如植物雌激素作为合成雌激素的替代品。
发明内容
技术问题
因此,鉴于上述问题提出了本发明,并且本发明的目的是提供一种含有葡萄籽提取物、缬草提取物和红花籽提取物的用于预防和缓解女性绝经期症状的食品组合物。
技术方案
根据本发明,上述和其它目的可通过提供用于预防和缓解女性绝经期症状的食品组合物来实现,所述组合物包含葡萄籽提取物、缬草提取物和红花籽提取物的组合作为活性成分。
对于本发明的用于预防和缓解女性更年期症状的食品组合物,所述组合优选地含有10至40重量%的葡萄籽提取物、10至50重量%的缬草提取物和30至70重量%的红花籽提取物。当满足该范围时,食品组合物可以表现出改善雌激素受体活性、抑制破骨细胞分化、增强成骨细胞活性和改善IGF-1和骨钙蛋白表达的作用。此外,与单一提取物相比,葡萄籽提取物、缬草提取物和红花籽提取物的组合可以表现出预防和缓解女性绝经期症状的协同效应。
在更年期中,卵巢中的卵泡消耗导致雌激素分泌的减少和在周围组织中雄激素向雌激素转化的增加。其特征还在于由于下丘脑-垂体轴的负反馈机制而导致的,雌激素水平降低并且促卵泡激素和促黄体生成素分泌增加。雌激素信号传递通过雌激素受体介导,雌激素受体包括位于细胞核的雌激素受体-α(雌激素受体-α),雌激素受体-β和位于细胞膜的GPR30(G蛋白偶联受体30)。
雌激素受体的分布取决于组织,并且雌激素受体分布在各种组织中。因此,更年期中雌激素的减少在分布有雌激素受体的各种组织中引起症状。用于缓解女性更年期症状的大多数功能性成分包括具有与雌激素相似结构并因此与雌激素受体结合以提供雌激素活性的植物雌激素成分。
由此,为了提供预防和缓解女性绝经期症状的效果,需要开发能有效改善雌激素受体活性、无毒且含有大量植物雌激素的食品。
因此,作为研究来自无毒且无副作用的天然物质的有效改善雌激素受体表达的食品成分的研究结果,获得葡萄籽提取物、缬草提取物和红花籽提取物的最佳混合比,并且由10至40重量%的葡萄籽提取物、10至50重量%的缬草提取物和30至70重量%的红花籽提取物组成的组合高度有效地预防和缓解女性绝经期症状。
对于本发明的用于预防和缓解女性绝经期症状的食品组合物,葡萄籽提取物、缬草提取物和红花籽提取物优选地用水、C1至C4低级醇或其混合物作为提取溶剂进行提取,更优选地用水或70%乙醇或其混合物作为提取溶剂进行提取。
对于本发明的用于预防和缓解女性绝经期症状的食品组合物,葡萄籽提取物、缬草提取物和红花籽提取物的提取例如为热水提取、冷提取、回流提取或超声提取。
对于本发明的用于预防和缓解女性绝经期症状的食品组合物,提取物优选地包括通过提取然后过滤并在减压或真空下浓缩而获得的浓缩物。提取物优选为通过冷冻干燥、热风干燥或喷雾干燥获得的粉末。
对于本发明的用于预防和缓解女性绝经期症状的食品组合物,相对于食品组合物的总重量,所述组合优选以0.01至50重量%的量存在。当所述组合以小于0.01重量%的量存在时,预防和缓解女性绝经期症状的效果不足,而当所述组合以高于50重量%的量存在时,与使用量相比效果低,因此导致经济效率低。
本发明的用于预防和缓解女性绝经期症状的食品组合物优选地包括选自肉类、谷类、含咖啡因的饮料、普通饮料、巧克力、面包、点心、糖果、披萨、果冻、面条、口香糖、冰淇淋、维生素复合物和其他健康食品补充剂中的任一种,但不发明不必限于此。
本发明的用于预防和缓解女性绝经期症状的食品组合物优选地进一步包括常用于食品生产的载体、赋形剂和稀释剂。这些成分的实例包括乳糖、右旋糖、蔗糖、山梨醇、甘露醇、木糖醇、赤藓糖醇、麦芽糖醇、淀粉、阿拉伯胶、藻酸盐、明胶、磷酸钙、硅酸钙、纤维素、甲基纤维素、微晶纤维素、聚乙烯吡咯烷酮、水、蜂蜜、羟基苯甲酸甲酯、羟基苯甲酸丙酯、硬脂酸镁和矿物油。使用选自常用填料、增稠剂、粘合剂、润湿剂、崩解剂和表面活性剂中的一种或多种稀释剂或赋形剂制备制剂。
同时,用于口服施用的固体制剂包括选自片剂、丸剂、粉末、颗粒和胶囊中的一种或多种,并且这些固体制剂例如通过将选自淀粉、碳酸钙、蔗糖或乳糖中的一种或多种赋形剂与明胶混合而制备。另外,除了简单的赋形剂之外,可以使用诸如硬脂酸镁等润滑剂。
另外,用于口服施用的液体制剂包括选自悬浮液、液体、乳液和浆液中的一种或多种,并且除了作为简单稀释剂的水和液体石蜡之外,还可以包括各种赋形剂,例如湿润剂、甜味剂、香料和补充剂。
本发明的具有预防和缓解女性绝经期症状的效果的食品组合物可进一步包括添加剂,其例如包括选自营养素、维生素、矿物质(电解质)、合成调味剂和天然调味剂、着色剂和填料(奶酪,巧克力)、果胶酸及其盐、藻酸及其盐、有机酸、保护性胶体增稠剂、pH调节剂、稳定剂、防腐剂、甘油、醇和用于碳酸饮料中的碳酸盐中的一种或多种。此外,食品组合物可以包括用于制备天然果汁、果汁饮品和水果饮料的果肉。这些成分可单独或以其组合使用。相对于100重量份的所述组合,添加剂一般以0至50重量份的量存在,但本发明不限于此。
对于具有预防和缓解女性绝经期症状的效果的食品组合物,“女性绝经期症状”意指在绝经期女性中由激素产生减少引起的症状,并且更年期或绝经期症状例如包括选自面部潮红、出汗、睡眠期间出汗、皮肤干燥、阴道干燥、阴道萎缩、下尿道萎缩、性交疼痛、阴道炎、膀胱炎、排尿困难、紧急排尿、注意力丧失、短期记忆失调、焦虑、神经过敏、记忆力减退、性欲减退症、肌痛、关节痛和骨质疏松症中的一种或多种。
同时,葡萄(Vitis vinifera L)是一种藤本果树,是世界上栽培最广泛的藤本植物,属于鼠李目(Rhamnales),葡萄科(Vitaceae),分为欧洲葡萄和美洲葡萄。欧洲葡萄含有大量的碳水化合物,包括作为主要糖的葡萄糖和少量其他糖。另外,欧洲葡萄含有少量的维生素B和C。其果汁含有酒石酸、苹果酸、柠檬酸、外消旋酸和水杨酸。葡萄具有健胃、利尿、滋补和止渴的功效。据报道,占葡萄重量3%至5%的葡萄籽在加工后含有作为副产物的生物活性物质,例如酚类化合物和儿茶素。特别是,近来有关儿茶素的活性氧清除的研究报道,其作为天然抗氧化剂使用的兴趣正在增加。
与此同时,被称为“Valeriana fauriei Briquet”的缬草(Valerianaofficinalis)是一种属于缬草科的多年生开花植物,并且产生高度约1至2米的深蓝色叶子和白色或粉红色花朵。缬草常见于欧洲和亚洲北部,据了解,其从古代起便已被用作药草,而它的根自公元前便已被使用。已知该植物的根和根茎提取物表现出各种效果,并且效果的代表性实例包括睡眠辅助、镇静作用、抗焦虑作用、抗惊厥作用和疼痛缓解。
同时,红花(Carthamus tinctoris)是属于菊科的一年生植物,在韩国,日本和中国主要栽培用于医学应用。据报道,红花籽由于大量脂肪尤其是亚油酸而表现出血液中胆固醇的降低。
有利效果
本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合具有抑制破骨细胞分化、增强成骨细胞活性、改善雌激素受体活性以及改善IGF-1和骨钙蛋白表达的效果,从而发挥预防和缓解女性绝经期症状的优异效果。另外,本发明提供具有预防和缓解女性绝经期症状的优异效果的食品组合物,其含有由无毒、无副作用且对人体无害的天然提取物组成的组合作为活性成分。
附图说明
根据以下结合附图的详细描述,将更清楚地理解本发明的上述和其他目的、特征和其他优点,其中:
图1是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对破骨细胞形成的影响的TRAP测量试验结果的图表(P<0.05);
图2是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对破骨细胞形成的影响的RANKL试验结果的图表(P<0.05);
图3是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对成骨细胞活性的影响的碱性磷酸酶(ALP)活性测量试验结果的图表(P<0.05);
图4是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对成骨细胞活性的影响的碱性磷酸酶(ALP)表达测量试验结果的图表(P<0.05);
图5是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对IGF-1基因表达的影响的试验结果的图表;
图6是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对雌激素受体表达的影响的试验结果的图表(P<0.05);并且
图7是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合对骨钙蛋白表达的影响的试验结果的图表(P<0.05)。
具体实施方式
在下文中,将参照以下实施例更详细地描述本发明。本发明的范围不限于以下实施例,并且包括与其等同的技术构思的变型。
[制备例1:葡萄籽提取物的制备]
将1kg葡萄籽干燥并研磨,将300g所得粉末置于气体收集瓶中,向其中加入1.8L的70%乙醇,并且在85℃重复进行两次3小时萃取,以获得葡萄籽提取物。然后,将提取物通过滤纸(Whatman No.1,England)在减压下过滤,在真空旋转蒸发器中于60℃下减压浓缩并喷雾干燥以制备葡萄籽提取物。
[制备例2:缬草提取物的制备]
将1kg缬草根干燥并研磨,将300g所得粉末置于气体收集瓶中,向其中加入1.5L的70%乙醇,并且在70℃重复进行三次6小时萃取,以获得缬草提取物。然后,将提取物通过滤纸(Whatman No.1,England)在减压下过滤,在真空旋转蒸发器中于60℃下减压浓缩并喷雾干燥以制备缬草提取物。
[制备例3:红花籽提取物的制备]
将1kg红花籽干燥并研磨,将300g所得粉末置于气体收集瓶中,向其中加入3L的70%乙醇,并且在90℃重复进行三次6小时萃取,以获得红花籽提取物。然后,将提取物通过滤纸(Whatman No.1,England)在减压下过滤,在真空旋转蒸发器中于60℃下减压浓缩并喷雾干燥以制备红花籽提取物。
[实施例1至4:组合的制备]
将制备例1至3中制备的葡萄籽提取物、缬草提取物和红花籽提取物以下表1中所示的不同重量比混合以制备组合。
表1
[试验例1:组合对抗酒石酸酸性磷酸酶(TRAP)活性——骨质疏松症指标的影响]
在本试验例中,鉴定了制备例1至3中制备的单个提取物和实施例1至4中制备的提取物组合对TRAP(抗酒石酸酸性磷酸酶)活性的影响。
抗酒石酸酸性磷酸酶(TRAP)是破骨细胞的指标。TRAP活性低意味着破骨细胞分化被抑制。
为了将MC3T3-E1细胞分化成破骨细胞,从7周龄的C57BL/6雌性小鼠中提取股骨骨髓细胞并与MC3T3-E1细胞在MEM(补充有10%FBS、100U/ml青霉素和100g/ml链霉素的最低必需培养基)中一起培养。
将MC3T3-E1细胞(1.5×104个细胞/孔)和骨髓细胞(5×105个细胞/孔)接种到24孔板上,并用10-8M 1,25(OH)2维生素D3、10-7M地塞米松和浓度为50μg/ml的样品(制备例1至3和实施例1至4)处理MEM培养基。每三天更换培养基,细胞培养七天。
培养七天后,将细胞用磷酸盐缓冲盐水(PBS)洗涤两次,通过用裂解缓冲液刮取收集,并以10秒的间隔超声处理4次以获得细胞裂解物。
本文使用的TRAP活性测量试剂盒是由Lifespan Biosciences生产的ELISA试剂盒,并且根据制造商的说明书在450nm下测量TRAP活性。
将未用1,25(OH)2维生素D3处理的组指定为一般组,将用1,25(OH)2维生素D3处理且未用样品进行处理的组指定为对照组。结果在下表2中示出。通过ANOVA分析获得试验组之间的显著差异,并且将p<0.05确定为显著差异。
表2
TRAP(相对于对照的%) | |
一般组 | 11±11.4d |
对照组 | 100.2±2.86a |
实施例1 | 81.6±4.51c |
实施例2 | 82.2±3.03c |
实施例3 | 89.6±4.28b |
实施例4 | 85.4±2.61bc |
制备例1 | 93.6±3.43ab |
制备例2 | 97.8±2.77a |
制备例3 | 94.6±2.61a |
试验结果显示了包含葡萄籽提取物、缬草提取物和红花籽提取物的组合表现出比单一提取物(葡萄籽提取物、缬草提取物和红花籽提取物)低的TRAP活性。另外,在所述组合中,实施例1(20重量%的葡萄籽提取物、30重量%的缬草提取物和50重量%的红花籽提取物)表现出最低的TRAP活性。
在以下试验例中,用发现具有最高效果的实施例1进行试验。
[试验例2:组合浓度对抗酒石酸酸性磷酸酶(TRAP)活性——骨质疏松症指标的影响]
在本试验例中,鉴定了组合浓度对TRAP活性的影响。
将MC3T3-E1细胞(1.5×104个细胞/孔)和骨髓细胞(5×105个细胞/孔)接种到24孔板上,并用10-8M 1,25(OH)2维生素D3、10-7M地塞米松以及0、20、50和100μg/ml的样品(实施例1)处理MEM培养基。每三天更换培养基,细胞培养七天。
培养七天后,将细胞用磷酸盐缓冲盐水(PBS)洗涤两次,通过用裂解缓冲液刮取收集,并以10秒的间隔超声处理4次以获得细胞裂解物。
本文使用的TRAP活性测量试剂盒是由Lifespan Biosciences生产的ELISA试剂盒,并且根据制造商的说明书在450nm下测量TRAP活性。将未用1,25(OH)2维生素D3处理的组指定为一般组,将未用组合物进行处理的组指定为对照组。
试验结果显示,当以50μg/ml和100μg/ml的浓度处理组合(实施例1)时,TRAP活性被浓度依赖性地显著抑制。
这些结果显示,根据组合的浓度,改善了抑制破骨细胞分化的效果(图1)。图1是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对破骨细胞形成的影响的TRAP测量试验结果的图表(P<0.05)。
[试验例3:组合对破骨细胞分化的影响的鉴定(RANKL表达的测量)]
在本试验例中,测量表达的RANKL(NF-κB配体的受体活化剂)的量以鉴定通过所述组合抑制破骨细胞分化的效果。
作为TNF家族的一部分,已知RANKL在破骨细胞分化中通过NF-κB和MAPK(丝裂原活化激酶)活化(J.Bone Miner Metab.2014.32:598-604),并充当破骨细胞分化的标记。
将MC3T3-E1细胞(1.5×104个细胞/孔)和骨髓细胞(5×105个细胞/孔)接种到24孔板上,并用10-8M 1,25(OH)2维生素D3、10-7M地塞米松以及0、20、50和100μg/ml的样品(实施例1)处理MEM培养基。每三天更换培养基,细胞培养七天。
培养七天后,除去细胞培养基,并根据制造商的说明书使用Quantikine M小鼠RANKL免疫测定试剂盒(R&D System,USA)进行试验。使用ELISA读取器在40分钟内于450nm处测量每个孔。将未用1,25(OH)2维生素D3处理的组指定为一般组,将未用组合物进行处理的组指定为对照组。
试验结果显示,当以50μg/ml和100μg/ml的浓度处理组合(实施例1)时,表达的RANKL的量被浓度依赖性地显著降低。
由这些结果可以确认,所述组合具有抑制破骨细胞分化的效果(图2)。图2是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对破骨细胞形成的影响的RANKL试验结果的图表(P<0.05)。
[试验例4:碱性磷酸酶(ALP)活性的测量]
成骨细胞在细胞膜上具有碱性磷酸酶(ALP),已知这种酶是成骨细胞的活性标记,其在细胞的外膜和钙化组织中以高浓度存在,并在钙化过程中调节转运、细胞分裂和分化(Int Rev Cytol.1995.265-358)。
因此,在本试验例中,测量碱性磷酸酶(ALP)的活性以鉴定组合对成骨细胞活性的影响。
为了测量ALP活性,将MC3T3-E1细胞系用MEM培养基、10mMβ-甘油磷酸酯和50μg/ml抗坏血酸处理6天,以使细胞分化为成骨细胞。将分化的细胞接种到24孔板上,用0、20、50和100μg/ml的所述组合(实施例1)处理,并培养3天。
培养后,将细胞用磷酸盐缓冲盐水(PBS)洗涤两次,通过用裂解缓冲液刮取收集,并以10秒的间隔进行超声处理4次。向每个孔中加入1ml反应混合物(碱性缓冲液:储备底物(stock substrate)为1:1)并在37℃温育30分钟。为了测量,根据Abcam方案进行试验。最后,向每个孔中加入12μL的1N NaOH,并使用ELISA读取器(VERSAMAXSL-20 MolecularDevices,Seoul,Korea)于450nm处测量吸光度。
试验结果显示,与未用所述组合处理的对照组相比,用不同浓度的所述组合处理的细胞表现出优异的ALP活性,特别是在以20、50和100μg/ml处理时浓度依赖性地表现出高ALP活性。
这些结果显示了所述组合具有促进成骨细胞活性的效果(图3)。图3是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对成骨细胞活性的影响的碱性磷酸酶(ALP)活性测量试验结果的图表(P<0.05)。
[试验例5:组合对碱性磷酸酶(ALP)、IGF-1、雌激素受体和骨钙蛋白的基因表达的影响]
IGF-1(胰岛素样生长因子-1)是一种与生长激素密切相关的生长因子,由通过生长激素刺激的脑垂体分泌。已知IGF-1的功能是促进软骨组织中软骨细胞的生长和分化并促进骨骼生长(Nat Med.2003.9:1033-1038)。
雌激素受体是一组蛋白质,其存在于各组织的核或其细胞中,其接受雌激素并进行雌激素信号传导。因此,雌激素在更年期中减少,并且分布有雌激素受体的各种组织中出现症状。通过测量雌激素受体的基因表达程度来评估组合的雌激素活性。
骨钙蛋白由成骨细胞合成并用作骨形成的指标。也就是说,血清中骨钙蛋白的浓度直接反映成骨细胞的活性。
因此,在本试验例中,进行实时PCR以鉴定组合对碱性磷酸酶(ALP)、IGF-1、雌激素受体和骨钙蛋白的基因表达的影响。
将MC3T3-E1细胞系用MEM培养基、10mMβ-甘油磷酸酯和50μg/ml抗坏血酸处理6天,以使细胞分化为成骨细胞。将分化的细胞接种到24孔板上,用0、20、50和100μg/ml的所述组合(实施例1)处理,并培养3天。
用0.25%胰蛋白酶-EDTA收集培养3天的细胞,根据制造商的说明书使用RNeasy提取试剂盒(QIAGEN,USA)提取总RNA,以便合成cDNA,将使用iScriptTM选择cDNA合成试剂盒(BIO-RAD,USA)向5μg的总RNA添加4μL 5×iScript选择反应混合物、2μL低聚(dT)引物组、5μL RNA样品和8μL无核酸酶水,最后,向其中加入1μL iScript逆转录酶,并通过用移液管上下摇动来充分混合混合物。
将反应在42℃进行60分钟,然后在85℃进行5分钟,然后将合成的cDNA用于PCR反应。对于实时PCR使用一步实时PCR系统(Applied Biosystems,USA),并根据iQ SYBR GreenSupermix(BIO-RAD,USA)的方法进行反应。
用于PCR的混合物的最终组成为2μL(10至100ng)的cDNA,10μL的2×iQ SYBRGreen Supermix,1μL(250nM)的各正向和反向引物,以及7μL的H2O。在95℃热启动10分钟后,通过在95℃下15秒、在55℃下15秒和在72℃下30秒循环40次进行PCR,最后是在95℃下15秒、在60℃下1分钟和在95℃下15秒的整理工序。用于反应的引物在下表3中示出。
表3
碱性磷酸酶(ALP)表达的测量结果显示,与对照组相比,以50μg/ml和100μg/ml的组合处理的细胞显示出碱性磷酸酶(ALP)的浓度依赖性显著增加(图4)。图4是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对成骨细胞活性的影响的碱性磷酸酶(ALP)表达测量试验结果的图表(P<0.05)。
IGF-1表达的测量结果显示,与对照组相比,以50μg/ml和100μg/ml的组合处理的细胞显示出IGF-1基因表达的浓度依赖性显著增加(图5)。图5是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对IGF-1基因表达的影响的试验结果的图表。
雌激素受体表达的测量结果显示,与对照组相比,以50μg/ml和100μg/ml的组合处理的细胞显示出雌激素受体表达的浓度依赖性显著增加(图6)。图6是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对雌激素受体表达的影响的雌激素受体表达测量试验结果的图表(P<0.05)。
骨钙蛋白表达的测量结果显示,与对照组相比,以20μg/ml、50μg/ml和100μg/ml的组合处理的细胞显示出骨钙蛋白表达的浓度依赖性显著增加(图7)。图7是示出鉴定本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)对骨钙蛋白表达的影响的试验结果的图表(P<0.05)。
由这些结果可以确认,所述组合具有增强成骨细胞活性和优异的雌激素活性的优异效果。
[实施例5:具有预防和缓解女性绝经期症状的效果的食品组合物的制备]
在本实施例中,制备了具有预防和缓解女性绝经期症状的效果的食品组合物。
(1)斋饭(Sunsik)(韩国传统谷物粉末混合物)的制备
将糙米、大麦、糯米和薏米糊化(α-转化),干燥,分散并使用研磨机制成粒度为60目的粉末。将黑豆、黑芝麻和紫苏通过熟知的方法蒸熟,干燥,分散并研磨以制备粒度为60目的粉末。然后,将30重量%的糙米、15重量%的薏米、20重量%的大麦、9重量%的糯米、7重量%的紫苏、8重量%的黑豆、7重量%的黑芝麻、3重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)、0.5重量%的灵芝(lacquer top)和0.5重量%的地黄混合以制备斋饭。
(2)口香糖的制备
将20重量%的胶基、76.9重量%的糖、1重量%的调味剂、2重量%的水和0.1重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)混合以通过常规方法制备口香糖。
(3)糖果的制备
将60重量%的糖、39.8重量%的淀粉糖浆、0.1重量%的调味剂和0.1重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)混合以通过常规方法制备糖果。
(4)饼干的制备
将25.59重量%的一等低筋面粉、22.22重量%的一等中筋面粉、4.80重量%的精制糖、0.73重量%的食盐、0.78重量%的葡萄糖、11.78重量%的棕榈起酥油、1.54重量%的铵、0.17重量%的碳酸氢钠、0.16重量%的亚硫酸氢钠、1.45重量%的米粉、0.0001重量%的维生素B1、0.0001重量%的维生素B2、0.04重量%的牛奶调味料、20.6998重量%的水、1.16重量%的全脂牛奶、0.29重量%的改性奶粉、0.03重量%的磷酸钙、0.29重量%的喷雾盐、7.27重量%的喷雾牛奶和1重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)混合以通过常规方法制备饼干。
(5)保健饮料的制备
将0.26重量%的蜂蜜、0.0002重量%的硫辛酸酰胺、0.0004重量%的烟酰胺、0.0001重量%的核黄素磷酸钠、0.0001重量%的盐酸吡哆醇、0.001重量%的肌醇、0.002重量%的乳清酸、98.7362重量%的水和1重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)混合以通过常规方法制备保健饮料。
(6)香肠的制备
将65.18重量%的猪肉、25重量%的鸡肉、3.5重量%的淀粉、1.7重量%的大豆蛋白、1.62重量%的食盐、0.5重量%的葡萄糖和1.5重量%的甘油与1重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)混合以通过常规方法制备香肠。
(7)健康补充食品的制备
将50重量%的本发明的包含葡萄籽提取物、缬草提取物和红花籽提取物的组合(实施例1)与16重量%的瓜尔胶水解物、0.01重量%的维生素B盐酸盐、0.01重量%的维生素B6盐酸盐、0.23重量%的DL-甲硫氨酸、0.7重量%的硬脂酸镁、31.2重量%的乳糖和1.85重量%的玉米淀粉混合以通过常规方法制备片剂型健康补充食品。
Claims (4)
1.一种预防和缓解女性绝经期症状的食品组合物,其包含葡萄籽提取物、缬草提取物和红花籽提取物的组合作为活性成分。
2.如权利要求1所述的食品组合物,其中所述组合是包含10至40重量%的所述葡萄籽提取物、10至50重量%的所述缬草提取物和30至70重量%的所述红花籽提取物的组合。
3.如权利要求1所述的食品组合物,其中所述葡萄籽提取物、所述缬草提取物和所述红花籽提取物利用水、C1至C4低级醇或其混合物作为提取溶剂进行提取。
4.如权利要求1所述的食品组合物,其中相对于所述食品组合物的总重量,所述组合以0.01至50重量%的量存在。
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2016
- 2016-01-12 KR KR1020160003956A patent/KR101653678B1/ko active IP Right Grant
- 2016-03-07 CN CN201680078067.4A patent/CN108471795A/zh active Pending
- 2016-03-07 WO PCT/KR2016/002224 patent/WO2017122868A1/ko active Application Filing
- 2016-03-07 JP JP2018535896A patent/JP6655189B2/ja not_active Expired - Fee Related
- 2016-03-07 EP EP16885180.6A patent/EP3403511A4/en not_active Withdrawn
- 2016-03-07 US US16/068,213 patent/US20190022163A1/en not_active Abandoned
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Also Published As
Publication number | Publication date |
---|---|
JP6655189B2 (ja) | 2020-02-26 |
KR101653678B1 (ko) | 2016-09-02 |
US20190022163A1 (en) | 2019-01-24 |
JP2019505208A (ja) | 2019-02-28 |
WO2017122868A1 (ko) | 2017-07-20 |
EP3403511A1 (en) | 2018-11-21 |
EP3403511A4 (en) | 2019-03-27 |
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