CN110538100A - 一种诃子醇提取物的应用 - Google Patents
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- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种诃子醇提取物的应用,具体为在制备抗皮肤衰老基因的表达上调剂中的应用。诃子醇提取物可以促进胞外基质合成和稳定基底膜结构完整,稳固皮肤结构的作用,诱导细胞修复相关的及抗衰老相关的基因表达,提高皮肤的结构的稳定和延缓衰老的功效;在特定的浓度范围下,可使抗皮肤衰老基因的表达量上调至1.37倍以上,最高可达2.2倍。该发现对于日后研发抗衰老相关的护肤品或者化妆品有重要意义。
Description
技术领域
本发明涉及植物提取物领域及日化领域,具体涉及一种诃子醇提取物的应用。
背景技术
皮肤覆盖于人体表面,与外界环境直接接触,除了能够保护机体免受外界各种因素的伤害,同时作为一面镜子反应机体的健康状态。皮肤由表皮、真皮、皮下组织构成,并含有皮肤附属器、丰富的神经、血管、淋巴管及肌肉。真皮和表皮的连接的细胞外基质区域又称为基底膜(Basement membrane,BM),不仅可以支撑表皮,而且具有多方面的功能,比如,真表皮连接处作为细胞的选择性通过屏障,通常可以阻止结缔组织细胞与基底膜的表皮细胞接触,但允许淋巴细胞、巨噬细胞和神经元突触穿越通过;此外,细胞的形态、细胞的极性、细胞代谢、质膜上蛋白质的分布、细胞存活、细胞的增殖分化、细胞迁移等许多生命现象,均与基底膜有着非常密切的关系。巢蛋白和基底膜聚糖是基底膜的重要组成部分,对维持基底膜的结构和功能必不可少,而基底膜的完整和健康对维持皮肤正常更新是非常必要的。而真皮由细胞和细胞外基质共同构成,成纤维细胞是真皮的主要细胞,其正常的增殖、分化维系着皮肤真皮结构、组成正常的结构和生理功能;细胞外基质是由细胞合成并分泌,围绕细胞相互作用的一类大分子物质,细胞的增殖分化、细胞迁移等许多生命现象,均与细胞外基质有着非常密切的关系。细胞外基质主要由四大类分子组成:胶原、弹性蛋白、糖蛋白及蛋白多糖。细胞外基质通过其分子间不同的精细巧妙缠结交织,构成各种各样的精美三维立体结构,赋予肌肤弹性、抗张强度,使肌肤看起来饱满、充满活力。
FGF7(Fibroblast growth factor 7)基因编码的成纤维细胞生长因子7促有丝分裂和细胞存活活性,并参与多种生物过程,包括胚胎发育,细胞生长,形态发生,组织修复,肿瘤生长和侵袭。这种蛋白质是一种有效的上皮细胞特异性生长因子,其促有丝分裂活性主要表现在角质形成细胞中,具有抗衰老的功效。
FBN1(Fibrillin 1)基因编码的原纤蛋白1是由真皮的成纤维细胞分泌到细胞外基质中的,可作为钙结合微原纤维的结构成分,为整个身体的弹性和非弹性结缔组织提供受力结构,同时介导一系列的细胞和基质的相互作用。是结缔组织中弹性纤维的重要成分,在弹性组织形成的过程中,原纤蛋白1为存放弹性蛋白原提供骨架,生长成熟弹性纤维的外围,皮肤弹性的下降是皮肤衰老的重要特征,原纤蛋白1的表达量与皮肤弹性息息相关,原纤蛋白1表达量的下降也是造成皮肤老化的关键因素。
CSF2(colony stimulating factor 2)基因编码的蛋白为集落刺激因子2,也称为粒细胞巨噬细胞集落刺激因子,是一种广谱的细胞因子,对早期的造血祖细胞具有促增殖的作用。主要作用于髓系祖细胞,通过与其高亲和力受体结合发挥生物学效应,使其迅速进入细胞周期向粒系和巨噬系分化,最后成为成熟细胞,并能延长成熟细胞的寿命,增强成熟细胞的功能,抵抗皮肤衰老,所以集落刺激因子2是一种重要的细胞长寿因子。
肝素结合表皮生长因子HBEGF(heparin binding EGF like growth factor)是结合并激活EGF受体生长因子的EGF家族的成员。HBEGF在病理生理过程中起关键作用,包括癌症。在人体皮肤的部分厚度损伤后,会产生表皮生长因子,促进细胞增殖,对皮肤伤口愈合有益,同时EGF是表皮生长因子,促进表皮细胞的增殖,维持皮肤年轻态。
TNC(tenascin C)基因编码的肌腱蛋白C,也称作细胞粘合素C(Tenascin-C,TNC)主要功能是加强结缔组织,在胚胎发育时期高表达,且被严格调控,而在正常成人组织中几乎不可检测到。成人组织TNC表达上调通常与炎症、损伤、肿瘤新生等疾病状态有关。TNC作为重要的非结构性细胞外基质蛋白,参与组织损伤修复过程,TNC表达水平的提高,可以反映出皮肤的增殖速率增加,可以帮组皮肤抵抗衰老。
LTBP1(latent transforming growth factor beta binding protein 1)基因编码的潜伏转化生长因子β结合蛋白1,属于潜伏的TGF-β结合蛋白(LTBP)家族。TGF-β的分泌和活化受其与潜伏相关蛋白和潜伏TGF-β结合蛋白的相关性调节。转化生长因子(transforming growth factor-β,TGF-β)是一类具有多效且功能复杂的细胞因子,参与细胞生长,组织修复和创面愈合,血管及瘢痕的生成及肿瘤的发生和转移。所以潜伏转化生长因子β结合蛋白1和皮肤伤口愈合,组织修复和抗衰老密切相关。
ITGB4(Intergrin-β4)基因编码的整合素β4亚基,整合素是细胞表面受体的主要家族,对细胞和细胞外基质的粘附起介导作用。整合素β4亚基与α6亚基结合,是基底膜中半桥粒的重要组成部分。整合素β4亚基定位在表皮基底层角质细胞中,是重要的细胞增殖能力标志,同时ITGB4基因表达上调能够促进整合素β4亚基的表达,进而促进基底膜结构的稳固性,强化真表皮的紧密连接,抵抗皮肤衰老。
因此,研究如何提高皮肤关键部位及功能相关的基因的表达,对于提高皮肤真表皮结构的稳定性,延缓皮肤衰老具有重要意义。
诃子,中药名,别名诃黎勒、诃黎、随风子。为使君子科植物诃子Terminaliachebula Retz.或绒毛诃子Terminalia chebula Retz.var.tomentella Kurt.的干燥成熟果实。分布于云南等地。乔木,高可达30米,径达1米,树皮灰黑色至灰色,粗裂而厚,枝无毛,皮孔细长,明显,白色或淡黄色;幼枝黄褐色,被绒毛。叶互生或近对生,叶片卵形或椭圆形至长椭圆形,长7-14厘米,宽4.5-8.5厘米,先端短尖,基部钝圆或楔形,偏斜,边全缘或微波状,两面无毛,密被细瘤点,侧脉6-10对;叶柄粗壮,长1.8-2.3厘米,稀达3厘米,距顶端1-5毫米处有2(-4)腺体。穗状花序腋生或顶生,有时又组成圆锥花序,长5.5-10厘米;花多数,两性,长约8毫米;花萼杯状,淡绿而带黄色,干时变淡黄色,长约3.5毫米,5齿裂,长约1毫米,三角形,先端短尖,外面无毛,内面被黄棕色的柔毛;雄蕊10枚,高出花萼之上;花药小,椭圆形;子房圆柱形,长约1毫米,被毛,干时变黑褐色;花柱长而粗,锥尖;胚珠2颗,长椭圆形。核果,坚硬,卵形或椭圆形,长2.4-4.5厘米,径1.9-2.3厘米,粗糙,青色,无毛,成熟时变黑褐色,通常有5条钝棱。花期5月,果期7-9月。药材性状为长圆形或卵圆形,长2-4cm,直径2-2.5cm。表面黄棕色或暗棕色,略具光泽,有5-6条纵棱线和不规则的皱纹,基部有圆形果梗痕。质坚实。果肉厚0.2-0.4cm,黄棕色或黄褐色。果核长1.5-2.5cm,直径1-1.5cm,浅黄色,粗糙,坚硬。种子狭长纺锤形,长约1cm,直径0.2-0.4cm,种皮黄棕色,子叶白色,相互重叠卷旋。气微,味酸涩后甜。
诃子出自《本草图经》,为清凉解毒中药,主治久泻、久痢、脱肛、喘咳痰嗽、久咳失音。明朝的《本草经疏》有如下记载“诃黎勒,其味苦涩,其气温而无毒。苦所以泄,涩所以收,温所以通,惟敛故能主冷气,心腹胀满;惟温故下食。甄权用以止水道,萧炳用以止肠僻久泄,苏颂用以疗肠风泻血、带下,朱震亨用以实大肠,无非苦涩收敛,治标之功也。”明末医书《药品化义》记载“诃子味苦而带酸涩,能降能收,兼得其善,盖金空则鸣,肺气为火邪郁遏,以致吼喘咳嗽,或至声哑,用此降火敛肺,则肺窍无壅塞,声音清亮矣。取其涩可去脱。若久泻久痢,则实邪去而元气脱,用此同健脾之药,固涩大肠,泻痢自止。”诃子是最常用的藏药。在藏药学经典著作《晶珠本草》里,诃子被称为“藏药之王”。蒙医经典论著《金光注释集》中,有如下记载:诃子其根部表苦味,其尖部表辛味,其中间的肉质部显酸味,其外皮则为涩味,而靠近其核的肉则呈甘味。而在其他蒙医著作中对诃子有如下的阐述,尖部之辛主治“赫依”、“巴达干”诸病,近核的肉质部其甘味可治“希拉”病,全部之味则可治多种因素引起之综合征。
发明内容
本发明要解决的技术问题是针对现有技术中缺乏用于提高皮肤抗衰老能力的物质的问题,提供一种诃子醇提取物的应用。本发明的诃子醇提取物可以提高抗皮肤衰老基因的表达,进而起到延缓人皮肤衰老的作用。
发明人付出创造性劳动后意外发现诃子具有提高基底膜相关基因和抗皮肤衰老基因表达的能力,稳固皮肤结构,调控真皮中胶原合成,适用于具有延缓衰老功效的化妆品中。
本发明主要通过以下技术手段解决上述技术问题:
本发明提供一种诃子醇提取物在制备抗皮肤衰老基因的表达上调剂中的应用。
其中,所述的抗皮肤衰老基因不仅仅局限于与皮肤衰老直接相关的基因,例如参与皮肤组织损伤修复的基因,还涉及与皮肤衰老间接相关的基因,例如提高其表达可以促进基底膜结构的稳固性并强化真表皮的紧密连接进而抵抗皮肤衰老的基底膜相关基因。所述的抗皮肤衰老基因优选FBN1基因、TNC基因、HBEGF基因、CSF2基因、FGF7基因、ITGB4基因和LTBP1基因中的一种或多种。
所述的诃子醇提取物可为本领域常规的诃子醇提取物,优选诃子C1~C5醇提取物;更优选优选诃子乙醇提取物,所述的乙醇优选70-95%乙醇。
所述的诃子醇提取物较佳地通过下述方法制得,所述的方法包括:
(1)粉碎诃子得粉末;所述的诃子较佳地为诃子全草;
(2)加入乙醇进行提取,过滤,得滤液;所述的提取的温度较佳地为室温,所述的室温为20-30℃;
(3)蒸发滤液,即得;所述蒸发的程度为蒸发至干。
为了能够更加有效地提取诃子中的有效成分:步骤(1)中所述粉末的目数优选20~60目;步骤(2)中所述的乙醇优选70%乙醇,所述乙醇添加量较佳地为所述粉碎后的诃子的体积的8-12倍,更佳地为10倍;步骤(2)中所述的提取的次数优选1次以上,更优选2次;其中所述的提取较佳地为超声提取,所述超声的频率较佳地为400-550W,更佳地为500W;所述超声的时间为25-40min,较佳地为30min。
步骤(2)中所述的过滤的方法较佳地为离心,所述离心的速度较佳地为6000-10000rpm,更佳地为8000rpm,所述离心的时间较佳地为8-12分钟,更佳地为10分钟。
在本发明一较佳实施例当中,所述的诃子醇提取物通过下述方法制得,所述的方法包括:
(1)将干燥的诃子全草粉碎,得粉末,所述粉末的目数为20目;
(2)加入10倍于所述粉末的体积的70%乙醇于室温下进行500W超声提取2次,每次30min,离心过滤后将滤液合并;所述离心的速度为8000rpm,所述离心的时间为10分钟;
(3)滤液旋转蒸发至干,即得。
利用上述方法制备得到的诃子醇提取物在所述的抗皮肤衰老基因的表达上调剂中的浓度较佳地为8~120μg/ml,更佳地为10~100μg/ml;其中,在针对HBEGF、CSF2和/或ITGB4基因的表达上调剂中的浓度较佳地为80~120μg/ml;更佳地为100μg/ml;在针对FBN1、TNC、FGF7和/或LTBP1基因的表达上调剂中的浓度较佳地为8~12μg/ml,更佳地为10μg/ml。
所述的抗皮肤衰老基因的表达上调剂适用于化妆品或者护肤品中。
在符合本领域常识的基础上,上述各优选条件,可任意组合,即得本发明各较佳实例。
本发明所用试剂和原料均市售可得。
本发明的积极进步效果在于:
诃子醇提取物可以促进胞外基质合成和稳定基底膜结构完整,稳固皮肤结构的作用,诱导细胞修复相关的及抗衰老相关的基因表达,提高皮肤的结构的稳定和延缓衰老的功效;在特定的浓度范围下,可使抗皮肤衰老基因的表达量上调至1.37倍以上,最高可达2.2倍。该发现对于日后研发抗衰老相关的护肤品或者化妆品有重要意义。
附图说明
图1为100μg/mL诃子醇提取物促进FBN1基因表达结果示意图。
图2为100μg/mL诃子醇提取物促进TNC基因表达结果示意图。
图3为10μg/mL诃子醇提取物促进HBEGF基因表达结果示意图。
图4为10μg/mL诃子醇提取物促进CSF2基因表达结果示意图。
图5为100μg/mL诃子醇提取物促进FGF7基因表达结果示意图。
图6为10μg/mL诃子醇提取物促进ITGB4基因表达结果示意图。
图7为100μg/mL诃子醇提取物促进LTBP1基因表达结果示意图。
具体实施方式
下面通过实施例的方式进一步说明本发明,但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法,按照常规方法和条件,或按照商品说明书选择。
实施例1诃子醇提取物的生产
取干燥的诃子30g,使用小型药材粉碎机(屹立4两中药粉碎机)粉碎至20目左右,粉碎,于室温下以10倍体积的70%乙醇500W超声提取2次,每次30min,离心(8000rpm,10min)滤液合并,滤液旋转蒸发至干,得到诃子醇提取物约16.9g。
实施例2皮肤模型的制备
从皮肤组织中分别提取人原代成纤维细胞(NHDF)及人原代角质细胞(NHEK)(提取方法参考文献:伍津津,朱堂友.皮肤组织工程[M].北京:人民军医出版社,2009:72-103)。将人原代成纤维细胞复苏,复苏过程为:将冷冻管从液氮中取出,迅速投入37℃水浴融化,离心机1000rpm/min离心10min。将复苏所得细胞使用DMEM培养基在37℃的二氧化碳培养箱中培养一周后,进行消化(将细胞从培养箱中取出,吸弃原培养基,加PBS15mL润洗细胞一次,加8mL 0.05%胰蛋白酶后,于37℃的二氧化碳培养箱中孵育8min,取出,超净台中加10mL含FBS的DMEM培养基,移液枪取出细胞于50mL离心管中,离心机中1000rpm,离心10min后弃上清,细胞待用)后与胶原混合进行真皮阶段培养,于37℃的二氧化碳培养箱中培养3~4天,然后将使用饲养层培养的人原代角质细胞,消化接种(5×104个细胞)到真皮阶段的皮肤模型表面,依次经过表皮浸没培养及气液相培养阶段,培养至18天得到可用于检测的3D皮肤模型(制备皮肤模型的具体步骤详见ANNIE F.BLACK,Ph.D.CHARBEL BOUEZ,al.Optimization and Characterization of an Engineered Human SkinEquivalent.TISSUE ENGINEERING Volume 11,Number 5/6,2005.Mary Ann Liebert,Inc.)。
在人原代角质细胞接种到真皮阶段的皮肤模型表面培养至形成3D皮肤模型的第3天至第18天的过程中,根据所使用培养基的不同进行分组,形成空白对照(NT)组、加入诃子醇提取物(100μg/mL)的阳性对照组和加入10μg/mL诃子醇提取物(10μg/mL)的阳性对照组。
其中,诃子醇提取物(100μg/mL),指所得诃子醇提取物在单位体积(mL)培养基中的量为100μg,约0.01wt%。
诃子醇提取物(10μg/mL),指所得诃子醇提取物在单位体积(mL)培养基中的量为10μg,约0.001wt%。
实施例3RNA的提取
剪取3D皮肤模型组织约2*3mm,放入2mL EP管中;加入1mL Trizol,加入2颗钢珠,用组织破碎仪匀浆,50Hz超声5min后;9000rpm、4℃条件下离心1min,将上清液转移到新的1.5mL EP管中;加入200μL氯仿,轻轻颠倒数次混匀,室温放置5min;9000rpm、4℃条件下离心15min;移转上层水相(约400μL)于新1.5mL EP管中,加入等体积的异丙醇,混匀,室温静置10min;9000rpm、4℃条件下离心10min;弃上清液后,沉淀用预冷的70%无水乙醇洗1次,8000rpm、4℃条件下离心5min;弃上清液后,沉淀空气干燥5~10分钟后,溶于30μL DEPC(焦碳酸二乙酯)水中;分光光度计测定RNA浓度。
实施例4基因表达分析及结果判定
根据QuantiGene R 2.0Plex Assay试剂盒进行RNA的纯化和基因定量分析,使用luminex平台对基因表达进行分析,结果详见图1~7。
结果判定:
在测定基因表达的试验中,如果相对于未添加提取物的实验组,基因的表达量明显升高,可推断诃子可以促进胞外基质合成和稳定基底膜结构完整,稳固皮肤结构的作用,诱导细胞修复相关的及抗衰老相关的基因表达,提高皮肤的结构的稳定和延缓衰老的功效。
1、诃子醇提取物能够提高胞外基质相关基因的表达
由图1可知,添加诃子醇提取物的阳性组(100μg/mL),与胞外基质相关的FBN1基因表达上调1.37倍,较空白对照提高37%,该基因编码合成的原纤蛋白1,是结缔组织中弹性纤维的重要成分,在弹性组织形成的过程中,原纤蛋白1为存放弹性蛋白原提供骨架,生长成熟弹性纤维的外围,增强和支持皮肤组织结构的稳固性。
2、诃子醇提取物能够提高抗衰老相关基因的表达
由图2可知,添加诃子醇提取物的阳性组(100μg/mL),与胞外基质相关的TNC基因表达上调1.85倍,较空白对照提高85%,该基因编码合成的肌腱蛋白C作为重要的非结构性细胞外基质蛋白,参与组织损伤修复和纤维化过程。以上结果证明该提取物可以修护皮肤损伤,抵抗皮肤衰老。
由图3可知,添加诃子醇提取物的阳性组(10μg/mL),在皮肤模型中与促进细胞增殖及皮肤伤口愈合的HBEGF基因表达上调2.2倍,较空白对照提高120%,说明该提取物有提高细胞修复能力促进创伤修复的作用。
由图4可知,添加诃子醇提取物的阳性组(10μg/mL),抗衰老基因CSF2基因表达上调2.07倍,较空白对照提高107%,该基因编码合成的集落刺激因子2,作用于髓系祖细胞,使其迅速进入细胞周期向粒系和巨噬系分化,最后成为成熟细胞,并能延长成熟细胞的寿命,增强成熟细胞的功能,可以延缓皮肤衰老。
由图5可知,添加诃子醇提取物的阳性组(100μg/mL),在皮肤模型中与抗衰来相关的FGF7基因表达上调1.84倍,较空白对照提高84%,该基因编码合成的成纤维细胞生长因子7促有丝分裂和细胞存活活性,其促有丝分裂活性主要表现在角质形成细胞中,具有抗衰老的功效。
3、诃子醇提取物能够提高基底膜相关基因表达
由图6可知,添加诃子醇提取物的阳性组(100μg/mL),在皮肤模型中与基底膜相关的ITGB4基因表达上调1.44倍,较空白对照提高44%,该基因编码合成的整合素β4亚基定位在表皮基底层角质细胞中,是重要的细胞增殖能力标志,同时ITGB4基因表达上调能够促进整合素β4亚基的表达,进而促进基底膜结构的稳固性,强化真表皮的紧密连接,抵抗皮肤衰老。
4、诃子醇提取物能够提高其他皮肤相关基因表达
由图7可知,添加诃子醇提取物的阳性组(100μg/mL),在皮肤模型中与皮肤其他相关基因ITGB4基因表达上调1.52倍,较空白对照提高52%,该基因编码合成的潜伏转化生长因子β结合蛋白1,属于潜伏的TGF-β(转化生长因子)结合蛋白(LTBP)家族,转化生长因子参与细胞生长,组织修复和创面愈合,血管及瘢痕的生成及肿瘤的发生和转移。所以潜伏转化生长因子β结合蛋白1和皮肤伤口愈合,组织修复和抗衰老密切相关。
Claims (10)
1.一种诃子(Terminalia chebula Retz.)醇提取物在制备抗皮肤衰老基因的表达上调剂中的应用。
2.如权利要求1所述的应用,其特征在于,所述的抗皮肤衰老基因选自FBN1基因、TNC基因、HBEGF基因、CSF2基因、FGF7基因、ITGB4基因和LTBP1基因中的一种或多种。
3.如权利要求1所述的应用,其特征在于,所述的诃子醇提取物为诃子C1~C5醇提取物;优选诃子乙醇提取物,所述的乙醇优选70-95%乙醇;
较佳地,所述的诃子醇提取物通过下述方法制得,所述的方法包括:
(1)粉碎诃子得粉末;所述的诃子较佳地为诃子全草;
(2)加入乙醇进行提取,过滤,得滤液;所述的提取的温度较佳地为室温,所述的室温为20-30℃;
(3)蒸发滤液,即得;所述蒸发的程度为蒸发至干。
4.如权利要求3所述的应用,其特征在于,步骤(1)中所述粉末的目数为20~60目。
5.如权利要求3所述的应用,其特征在于,步骤(2)中所述的乙醇为70%乙醇,所述乙醇添加量为所述粉碎后的诃子的体积的8-12倍,较佳地为10倍;
和/或,步骤(2)中所述的提取的次数为1次以上,较佳地为2次。
6.如权利要求3所述的应用,其特征在于,步骤(2)中所述的提取为超声提取,所述超声的频率较佳地为400-550W,更佳地为500W;
和/或,所述超声的时间为25-40min,较佳地为30min。
7.如权利要求3所述的应用,其特征在于,步骤(2)中所述的过滤的方法为离心,所述离心的速度较佳地为6000-10000rpm,更佳地为8000rpm,所述离心的时间较佳地为8-12分钟,更佳地为10分钟。
8.如权利要求3所述的应用,其特征在于,所述的诃子醇提取物通过下述方法制得,所述的方法包括:
(1)将干燥的诃子全草粉碎,得粉末,所述粉末的目数为20目;
(2)加入10倍于所述粉末的体积的70%乙醇于室温下进行500W超声提取2次,每次30min,离心过滤后将滤液合并;所述离心的速度为8000rpm,所述离心的时间为10分钟;
(3)滤液旋转蒸发至干,即得。
9.如权利要求8所述的应用,其特征在于,所述的诃子醇提取物在所述的抗皮肤衰老基因的表达上调剂中的浓度为8~120μg/ml,较佳地为10~100μg/ml;其中,在针对HBEGF、CSF2和/或ITGB4基因的表达上调剂中的浓度较佳地为80~120μg/ml;更佳地为100μg/ml;在针对FBN1、TNC、FGF7和/或LTBP1基因的表达上调剂中的浓度较佳地为8~12μg/ml,更佳地为10μg/ml。
10.如权利要求1~9任一项所述的应用,其特征在于,所述的抗皮肤衰老基因的表达上调剂适用于化妆品或者护肤品中。
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WO1998027956A1 (fr) * | 1996-12-20 | 1998-07-02 | Parfums Christian Dior | Utilisation d'un extrait de terminalia catappa |
KR20030086776A (ko) * | 2002-05-07 | 2003-11-12 | 주식회사한국신약 | 세포에 대한 노화 억제 작용을 하는 가자 추출물 및 이를함유하는 화장품 |
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