CN111315353A - 包含粗齿绣球提取物的因紫外线引起的人皮肤细胞损伤改善用组合物 - Google Patents
包含粗齿绣球提取物的因紫外线引起的人皮肤细胞损伤改善用组合物 Download PDFInfo
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- CN111315353A CN111315353A CN201880070370.9A CN201880070370A CN111315353A CN 111315353 A CN111315353 A CN 111315353A CN 201880070370 A CN201880070370 A CN 201880070370A CN 111315353 A CN111315353 A CN 111315353A
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Abstract
本发明涉及包含粗齿绣球提取物作为有效成分的因紫外线引起的人表皮细胞及真皮细胞损伤改善用组合物。根据本发明,本发明的组合物不仅可恢复人表皮细胞及真皮细胞中的因UV‑B引起的细胞损伤,而且增加透明质酸的分泌,并抑制MMP‑1的分泌,由此可改善因UV‑B引起的细胞损伤。因此,作为安全的原材料的粗齿绣球提取物可有用地使用于食品组合物领域或化妆品领域。
Description
技术领域
本发明涉及因紫外线引起的人皮肤细胞损伤改善用组合物,更具体地涉及包含粗齿绣球提取物作为有效成分的由UV-B暴露而产生的人表皮细胞及真皮细胞损伤的组合物。
背景技术
皮肤老化根据因素可划分为内源性老化(Intrinsic aging)和外源性老化(Extrinsic aging)。据悉,内源性老化的原因在于,随着年龄而出现的皮肤表皮及真皮的生理功能变化,外源性老化的原因在于,大气污染、紫外线暴露、应激等环境中出现的皮肤的生理功能变化。这种老化的机制中,被UV诱导的氧化应激(Oxidative Stress)增加体内的自由基(Free Radical),分解胶原蛋白(Collagen)的基质金属蛋白酶-1(MMP-1,matrixmetalloproteinase-1)的活性增加及分解透明质酸(Hyaluronic acid)的透明质酸酶(Hyaluronidase)的活性增加,由此引起皮肤表皮及真皮损伤。
在日常生活中,我们经常暴露在阳光中,尤其因UV-B暴露而产生的细胞的损伤及皮肤癌诱发是日常生活中不可避免的因素。
由此,活跃进行有关抑制因UV暴露而引起的皮肤细胞损伤的机制研究,并活跃进行有关呈现生化功效、效果的化妆品及保健品、医药品的研究。通过研究水准的发达及技术力增加,具有实质性功效、效果的材料为主要研究对象。
植物来源材料在安全性方面上优秀,并且长期被利用,尤其,对于韩国而言,活跃开发出将在民间利用或者在中药配方中利用的植物及生药成分作为主要成分的功能性材料。
粗齿绣球(Hydrangea serrata)为虎耳草科的宽叶灌木,在食品医药品安全厅中公告的食品原材料中的叶子部位被明示为可食用部位。中药配方中,生药名为七叶胆,其自古以来使用于慢性支气管炎、止咳、祛痰、消炎解毒等的治疗。在韩国公开专利第2004-0063874号中揭示含有具有高血糖及高脂血症抑制活性的甘茶粗齿绣球提取物的组合物。
但是,目前未直接进行有关粗齿绣球抑制因紫外线引起的人表皮细胞及真皮细胞的机制研究。因此,本发明人直接执行有关上述提取物具有的因紫外线引起的抗氧化酶活性和人表皮细胞及真皮细胞损伤抑制的基础功效研究。
对此,本发明人为了克服上述现有技术的问题,锐意研究努力的结果,含有粗齿绣球(Hydrangea serrata)的提取物作为有效成分的人表皮细胞及真皮细胞中通过抗氧化酶活性的ROS去除以及抑制因UV-B引起的皮肤损伤,由此确认可呈现皮肤改善效果,从而完成本发明。
现有技术文献
专利文献
(专利文献1)KR10-2004-0063874 A
(专利文献2)KR10-2005-0056569 A
非专利文献
(非专利文献1)WOO,M.S.,et al.Comparison of skin elasticity testresults from the Ballistometer and Cutometer Skin Research and Technology,2014,20.4:422-428.
(非专利文献2)IMOKAWA,Genji.Mechanism of UVB-induced wrinkling of theskin:paracrine cytokine linkage between keratinocytes and fibroblasts leadingto the stimulation of elastase.In:Journal of Investigative DermatologySymposium Proceedings.Nature Publishing Group,2009.p.36-43.
发明内容
技术问题
因此,本发明的主要目的在于,提供含有粗齿绣球提取物作为有效成分的因紫外线引起的人表皮细胞及真皮细胞损伤改善用组合物。
技术方案
根据本发明的一实施方式,本发明提供含有粗齿绣球(Hydrangea serrata)的提取物作为有效成分的因紫外线引起的人皮肤细胞损伤预防及改善用组合物。
本发明提供因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,上述因紫外线引起的人皮肤细胞损伤为因UV-B暴露而引起的人表皮细胞及真皮细胞损伤。对于本发明的实验例而言,在人表皮细胞及真皮细胞中不仅恢复因UV-B而引起的细胞损伤,而且增加透明质酸的分泌,并抑制MMP-1的分泌,由此证明具有因UV-B而引起的细胞损伤恢复效果。
在本发明中,人皮肤细胞的“损伤”包括因紫外线引起的人皮肤细胞的凋亡、皮肤细胞DNA损伤、活性氧增加、脂质过氧化增加等,其症状可包括红斑、日光火伤、色素沉着、光老化、皮肤癌等。并且,损伤的“预防”是指抑制或延迟上述因紫外线引起的皮肤细胞的损伤的所有行为。并且,损伤的“改善”是指缓解上述因紫外线引起的皮肤细胞的损伤状态或者减少症状的程度的所有行为。
本发明的特征在于,上述提取物能够由为了提取天然植物而利用的任何溶剂进行提取,尤其由水、碳原子数1至4的乙醇或它们的混合溶剂进行提取。上述醇可使用选自由乙醇、甲醇、异丙醇及丁醇等组成的组中的任一种以上,优选地可使用乙醇,更优选地可使用发酵酒精。
本发明的特征在于,上述提取物,相对于组合物总重量,可包含为0.001至80.0重量百分比,优选地包含为5至20重量百分比。并且,在上述条件下可确认高收率和从UV-B中恢复的显著功效。
在本发明的一实施例中,上述紫外线可以为UV-B。
根据本发明的实验例,可知粗齿绣球乙醇提取物可改善皮肤被UV-B暴露而出现的损伤。(参照实验例1、2)。
本发明的特征在于,上述组合物具有选自由片剂、颗粒剂、丸剂、胶囊剂、液态型、啫喱或口香糖组成的组中的一种以上的剂型,其为口腔摄取用。
本发明的特征在于,上述组合物具有选自由柔软化妆水、营养化妆水、营养霜、保湿霜、祛斑霜、凝胶乳液或软膏组成的组中的一种以上的剂型,其为皮肤外用剂用。
包含本发明的粗齿绣球提取物作为有效成分的组合物,可使用为多种用途,例如可使用为化妆品组合物、药学组合物、保健食品组合物等。
本发明的化妆品组合物还可包含配合于一般皮肤化妆品的化妆品学上可允许的一种以上载体,作为常规的成分,例如可适当配合油分、水、表面活性剂、保湿剂、低级醇、增粘剂、螯合剂、色素、防腐剂、香料等,但是并不局限于此。
本发明的药学组合物或保健食品组合物还可包含药学组合物的制备中通常使用的适当的载体、赋形剂或稀释剂等。上述药学上可允许的载体、赋形剂或稀释剂可包含选自由乳糖、葡萄糖、蔗糖、硅酸钙、纤维素、甲基纤维素、微晶纤维素、聚乙烯吡咯烷酮、水、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁及矿物油等组成的组中的任一种以上,但是不局限于此。
本发明的药学组合物的药学有效量、有效给药量可根据药学组合物的制剂化方法、给药方式、给药时间和/或给药途径等而多样。并且,上述药学组合物的给药,根据要实现的反应的种类与程度、作为给药对象的个体的种类、年龄、体重、一般的健康状态、疾病的症状或程度、性别、饮食、排泄、同时或异时一起使用于相关个体的药物其他组合物的成分等包括在内的多种因素及医药领域中公知的类似因素而多样。在该技术领域中普通技术人员可容易确定而处方对所目的的治疗有效的给药量。本发明的药学组合物,一天可给药1次,可分几次给药。因此,上述给药量在任何方面上限定本发明的范围。本发明的药学组合物的优选给药量可以是1天1mg/kg至1000mg/kg。
发明的效果
如上所述,包含本发明的粗齿绣球提取物的组合物,促进超氧化物歧化酶(SOD,Superoxide dismutase)的活性,在皮肤细胞中抑制因UV产生的活性氧(ROS),并且,恢复因UV产生的细胞损伤,由此适合作为用于皮肤美容的组合物。并且,包含本发明的粗齿绣球提取物的组合物增加透明质酸的分泌,抑制MMP-1的分泌,由此可防止因紫外线引起的皮肤老化,并可维持皮肤弹性。粗齿绣球提取物作为安全的材料可有用地使用于食品组合物领域或化妆品领域。
具体实施方式
以下,通过实施例更详细地说明本发明。由于这些实施例仅用于例示本发明,故而本发明的范围不得被解释为局限于这些实施例。
实施例1:利用乙醇的提取物的制备
本发明的组合物中所含的粗齿绣球提取物通过如下过程制备。首先,购买在济州岛内自生的1年生粗齿绣球,用蒸馏水洗净之后,在没有阳光的阴凉的地方完全干燥至无重量变化为止。之后,利用均质机粉碎完全干燥的干燥物,获得粉碎物之后,以7:3的重量比混合乙醇:蒸馏水,准备提取溶剂各400g之后,添加上述粉碎物50g。此时,粉碎物和提取溶剂的混合比率的重量比为1:8。接着,将与上述粉碎物混合的提取溶剂放入到振荡恒温水槽中,在50℃下搅拌提取2小时。通过提取获得的物质,在常温下使用沃特曼(No 2.)提取溶剂滤纸,去除不溶性物质。之后,在具有冷却冷凝器的蒸馏装置中进行减压浓缩,由此完全去除溶剂。完全干燥粗齿绣球提取物之后,测定提取收率的结果显示20%的收率。
实验例1:粗齿绣球乙醇提取物的细胞毒性评价
测定上述实施例1中获得的提取物是否对皮肤细胞具有细胞毒性。在本实验中,作为角质细胞使用HaCaT角质细胞(HaCaT Keratinocyte),作为真皮细胞使用HS68成纤维细胞(HS68 fibroblast)。为了确认是否有细胞毒性,将1.0X104细胞数的各个细胞分注于96-孔板上,粘附24小时,进行稳定化之后,以0、5、25、125ppm(μg/ml)的浓度处理粗齿绣球乙醇提取物。经过24小时之后,通过MTT试验,测定细胞存活程度。MTT试验是测定从被存活的细胞的线粒体还原的MTT形成的甲臜的量的方法。用5mg/ml MTT溶液50μl处理各个细胞,培养4小时之后,完全去除溶液,在540nm的吸光度下测定由二甲基亚砜(DMSO)溶解的板。
表1
如上述表1的结果所示,按不同浓度处理粗齿绣球提取物,执行细胞毒性试验,其结果确认真皮细胞及表皮细胞没有细胞毒性。
实验例2:粗齿绣球乙醇提取物的因UV-B引起的细胞损伤恢复评价
针对上述实施例1中获得的提取物,测定是否可恢复暴露在UV-B的皮肤细胞的损伤。在本实验中,作为角质细胞使用HaCaT Keratinocyte,作为真皮细胞使用HS68fibroblast。为了确认是否有细胞恢复功效,将1.0X104细胞数的各个细胞分注于96-孔板上,粘附24h,进行稳定化之后,以15mJ/cm2照射UV-B。之后,更换为以0、5、25、125ppm(μg/ml)的浓度处理粗齿绣球乙醇提取物的培养基,通过MTT试验测定细胞存活程度。MTT试验是测定从被存活的细胞的线粒体还原的MTT形成的甲臜的量的方法。更详细地,用5mg/ml MTT溶液50μl处理各个细胞,培养4小时之后,完全去除溶液,在540nm的吸光度下测定由DMSO溶解的板。
表2
P值(P-value):未用通过t-test的提取物处理的组(0μg/ml)和用提取物处理的组之间的统计处理结果如上述表2的结果所示,15mJ/cm2的UV-B处理时表皮细胞及真皮细胞中分别出现80%、76%的细胞损伤。对此,按不同浓度,用粗齿绣球提取物处理,执行因UV引起的皮肤细胞损伤恢复实验的结果,统计学上显著地可确认(P-value<0.05)细胞的损伤得到恢复。当考虑到本发明的组合物为提取物时,可确认浓度依赖性地较高的恢复功效。
实验例3:透明质酸(Hyaluronic Acid)含量分析
针对上述实施例1中获得的提取物,测定在真皮细胞HS68 fibroblast中是否可增加透明质酸。为了确认根据物质处理是否增加透明质酸,将1.0X105细胞数的各个细胞分注于24-孔板上,粘附24h,进行稳定化之后,替换为以0、5、25、125ppm(μg/ml)的浓度处理粗齿绣球乙醇提取物的培养基,培养24小时。之后,用物质处理的上清液中,使用透明质酸加酶联免疫吸附测定试剂盒(Hyaluronic Acid PLUS ELISA Kit)(TE 1018-2,TECO医务组(TECO medical Group,美国)测定透明质酸的分泌程度。
表3
P-value:未用通过t-test的提取物处理的组(0μg/ml)和用提取物处理的组之间的统计结果如上述表2的结果所示,按不同浓度,用粗齿绣球提取物处理,执行透明质酸实验的结果,在真皮细胞中处理25、125μg/ml的组中统计学上显著地确认(P-value<0.05)透明质酸的分泌促进效果。
实验例4:MMP-1的含量分析
针对上述实施例1中获得的提取物,测定在表皮细胞HaCaTKeratinocyte中是否可增加MMP-1。为了确认根据物质处理是否减少MMP-1,将1.0X105细胞数的各个细胞分注于24-孔板上,粘附24h,进行稳定化之后,替换为以0、5、25、125ppm(μg/ml)的浓度处理粗齿绣球乙醇提取物的培养基,培养24小时。之后,用物质处理的上清液中,使用人基质金属蛋白酶-1酶联免疫吸附测定试剂盒(MMP-1Human ELISA Kit)(ab100603,艾博抗(abcam),美国)测定MMP-1的分泌程度。
表4
P-value:未用t-test的提取物处理的组(0μg/ml)和用提取物处理的组之间的统计结果如上述表4的结果所示,按不同浓度,用粗齿绣球提取物处理,执行MMP-1实验的结果,在表皮细胞中统计学上显著地确认(P-value<0.05)MMP-1的分泌抑制效果。
剂型例1:片剂的制备
针对上述实施例1中获得的提取物,根据常规的片剂制备方法,混合下列表5的成分并进行压片,制备片剂。
表5
原料名 | 单位重量(mg) |
实施例1的提取物 | 50 |
玉米淀粉 | 100 |
乳糖 | 100 |
硬脂酸 | 2 |
剂型例2:胶囊剂的制备
针对上述实施例1中获得的提取物,根据常规的胶囊剂制备方法,混合下列表6的成分,填充在明胶胶囊中,制备胶囊剂。
表6
原料名 | 单位重量(mg) |
实施例1的提取物 | 50 |
玉米淀粉 | 100 |
乳糖 | 100 |
硬脂酸 | 2 |
剂型例3:液剂的制备
针对上述实施例1中获得的提取物,根据适合于嗜好的饮料制备方法,混合下列表7的成分,填充在瓶或袋中,制备液剂。
表7
剂型例4:啫喱的制备
针对上述实施例1中获得的提取物,根据适合于嗜好的啫喱制备方法,混合下列表8的成分,填充在三面布中,制备啫喱。
表8
原料名 | 单位重量(g) |
实施例1的提取物 | 2.0000 |
食用凝胶 | 0.3600 |
卡拉胶 | 0.0600 |
乳酸钙 | 0.1000 |
柠檬酸钠 | 0.0600 |
复合黄芩提取物 | 0.0200 |
酶改质甜菊糖 | 0.0440 |
低聚果糖液 | 5.0000 |
红葡萄浓缩液 | 2.4000 |
纯化水 | 13.9560 |
剂型例5:营养霜的制备
针对上述实施例1中获得的提取物,根据常规方法,以下列表9的组成,制备营养霜。
表9
上述的组成比,通常混合合适的成分,组成为剂型例,但是根据需要,还可任意变更其配合比及原料也无妨。
由于本发明的提取物在所有剂型例试验条件下稳定,因此在剂型的稳定性上无问题。
Claims (6)
1.一种因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,包含粗齿绣球的提取物作为有效成分。
2.根据权利要求1所述的因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,所述因紫外线引起的人皮肤细胞损伤为由UV-B暴露而产生的人表皮细胞及真皮细胞损伤。
3.根据权利要求1所述的因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,所述提取物由水、碳原子数1至4的醇或它们的混合溶剂进行提取。
4.根据权利要求1所述的因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,所述提取物相对于组合物总重量,包含为0.001至80重量百分比。
5.根据权利要求1所述的因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,所述组合物具有选自由片剂、颗粒剂、丸剂、胶囊剂、液态型、啫喱或口香糖组成的组中的一个以上的剂型,其为口腔摄取用。
6.根据权利要求1所述的因紫外线引起的人皮肤细胞损伤预防及改善用组合物,其特征在于,所述组合物具有选自由柔软化妆水、营养化妆水、营养霜、保湿霜、祛斑霜、凝胶乳液或软膏组成的组中的一个以上的剂型,其为皮肤外用剂。
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