CN110741074B - 具有促进毛发生长活性的弯曲乳杆菌wikim55和包含其的组合物 - Google Patents
具有促进毛发生长活性的弯曲乳杆菌wikim55和包含其的组合物 Download PDFInfo
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- CN110741074B CN110741074B CN201880036336.XA CN201880036336A CN110741074B CN 110741074 B CN110741074 B CN 110741074B CN 201880036336 A CN201880036336 A CN 201880036336A CN 110741074 B CN110741074 B CN 110741074B
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- lactobacillus curvatus
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Abstract
本公开涉及从泡菜分离的新型弯曲乳杆菌WIKIM55、及包含其的组合物。根据本公开的弯曲乳杆菌WIKIM55是具有促进毛发生长活性的益生菌,并且可以多样化地应用于人或动物中的肠道调节和促进毛发生长。
Description
技术领域
本公开涉及新型弯曲乳杆菌(Lactobacillus curvatus)菌株和包含其的组合物。
背景技术
毛发是一种身体器官,起到保护头部、保持头部温度等作用,并且决定外貌,从而引起了人们的极大兴趣。因此,对毛发脱落和毛发脱落治疗的兴趣正在增加,并且人们积极地进行对毛发脱落治疗的研究和开发。毛发在以下4个阶段的毛发周期内生长和脱落:毛发生长初期(生长期)、毛发生长中期(过渡期)、毛发生长终期(休止期)和毛发脱落期(脱落期)。毛发脱落是指由于各种原因,当毛发脱落超过毛发生长时引起的毛发脱落。当前已知的毛发脱落的原因包括遗传因素、血流问题、由雄性激素睾酮引起的内分泌失调等。此外,压力被认为是外部原因。
作为典型的毛发生长促进剂,使用了米诺地尔(6-氨基-1,2-二氢-1-羟基-2-亚氨基-4-苯氧基嘧啶,经皮搽剂)、非那雄胺(口服剂)等。已知米诺地尔可通过血管扩张促进营养供应,从而诱导毛发生长。非那雄胺具有通过抑制一种参与睾酮代谢的酶5α-还原酶的活性来抑制真皮乳头细胞死亡的作用机制。但是,这两种药物仅在使用过程中才有效,如果停止使用,效果会消失。因此,它们必须长时间使用。但是,由于米诺地尔具有黏腻感和皮肤刺激性的副作用,非那雄胺具有精力下降、性功能障碍等副作用,因此它们的长期使用存在许多限制。
在这种背景下,近来正在进行使用乳酸菌对皮肤和毛发健康的研究。但是,尚未发现直接影响毛发生长的乳酸菌,并且需要开发包含能够实现毛发健康和毛发生长的优异乳酸菌的组合物。
发明内容
技术问题
本公开内容旨在提供乳杆菌属中的新型乳酸菌,其具有促进毛发生长的优异活性。
技术方案
本公开的发明人致力于寻找一种乳酸菌菌株,其显示出促进毛发生长的作用,同时作为来自常规发酵食品的益生菌表现出优异的作用。因此,已经分离并鉴定了乳杆菌属的新型乳酸菌菌株,弯曲乳杆菌WIKIM55,并完成了本公开。
因此,本公开内容提供了用于促进毛发生长的药物组合物和食品组合物,其包含弯曲乳杆菌WIKIM55或其培养物作为活性成分;用于促进毛发生长的方法,该方法包括向有需要的对象施用治疗有效量的弯曲乳杆菌WIKIM55或其培养物;以及弯曲乳杆菌WIKIM55用于促进毛发生长的新用途。
根据本公开的弯曲乳杆菌WIKIM55是源自泡菜(kimchi)的新型弯曲乳杆菌菌株。尽管根据本公开的弯曲乳杆菌WIKIM55是从泡菜分离的,但是其来源不限于此。
在本公开的一个实例中,发现从常规发酵食品中分离的显示出作为益生菌的优异效果的乳酸菌菌株在进行用于对微生物进行鉴定和分类的16S rDNA序列分析时具有SEQID NO:1的核酸序列。
具有SEQ ID NO:1的16S rDNA碱基序列的微生物被命名为弯曲乳杆菌WIKIM55,并于2017年4月7日保藏在韩国微生物保藏中心(保藏号:KCCM12011P)。
本公开的弯曲乳杆菌WIKIM55是革兰氏阳性细菌。它是一种杆状兼性厌氧菌,能够在有氧和缺氧条件下生长而不形成孢子。
本公开的弯曲乳杆菌WIKIM55是益生菌,并且具有普通乳酸菌的肠道调节和免疫增强作用。众所周知,乳杆菌属中的乳酸菌具有缓解肠道疾病和增强免疫力的作用。
在本公开中,“益生菌”被理解为是指向宿主提供健康益处的活微生物,其改善包括人类在内的动物的胃肠道中的肠道微生物环境。益生菌是具有益生菌活性的活微生物,当以单个或多个菌株的形式提供给人或动物时,可以对宿主的肠道菌群产生有益作用。
在以下实例中,证实了本公开的弯曲乳杆菌WIKIM55菌株表现出通过促进毛囊形成来促进毛发生长的作用。因此,根据本公开的弯曲乳杆菌WIKIM55可用于人类或动物中的各种用途,例如肠道调节、促进毛发生长、增强免疫力等。
在本公开中,“促进毛发生长”的作用是指促进毛发的生长并最终增加相对于全部毛发,处于毛发生长初期的毛发的比例。因此,该术语是指抑制由处于毛发生长初期的毛发比例降低所引起的毛发脱落,并且可以与“毛发脱落改善”、“毛发脱落预防”和“毛发脱落治疗”具有相同的含义。
因此,本公开的示例性实施方案提供了包含弯曲乳杆菌WIKIM55或其培养物的益生菌组合物。
本公开的组合物中所含的弯曲乳杆菌WIKIM55可以以活细胞或死细胞的形式存在,也可以以干燥或冻干的形式存在。适用于包含在各种组合物中的乳酸菌的形式及其配制方法是本领域技术人员众所周知的。
在一个示例性的实施方案中,组合物可以是用于经口施用的组合物,其包含作为活细胞存在的弯曲乳杆菌WIKIM55菌株。
在另一个示例性的实施方案中,组合物可以是用于外部施用于皮肤的组合物,其包含弯曲乳杆菌WIKIM55菌株的培养物或裂解物或使用该菌株的发酵产物。
在一个示例性的实施方案中,本公开提供了肠道调节组合物,其包含弯曲乳杆菌WIKIM55或其培养物。根据本公开的肠道调节组合物可以用于预防、治疗和改善包括人在内的动物的胃肠道疾病。具体地,动物包括农场动物,如牛、马和猪。“胃肠道疾病”包括影响胃的细菌感染和炎性肠病。例如,包括但不限于由病原菌(大肠杆菌(E.coli)、沙门氏菌(Salmonella)、梭状芽孢杆菌(Clostridium)等)引起的感染性腹泻、胃肠道炎症、炎性肠病、神经源性肠炎综合征、小肠细菌过度生长综合征、肠道痢疾性腹泻等。
具体地,根据本公开的肠道调节组合物可以经口施用。给药剂量可以根据胃肠道疾病的类型、疾病的严重程度、年龄、性别、种族、治疗方法或预防而变化。通常,每天可能要向成年人施用一千至一千亿个细菌。
本公开提供了用于增强免疫力的组合物,其包含弯曲乳杆菌WIKIM55或其培养物。众所周知,乳杆菌属中的乳酸菌具有肠道调节和增强免疫力的作用。
本公开提供了用于促进毛发生长的组合物,其包含弯曲乳杆菌WIKIM55或其培养物。
由于这些有益效果,根据本公开的弯曲乳杆菌WIKIM55可包含在药物、保健食品、食品、饲料、饲料添加剂、乳酸发酵剂或化妆品中。
在一个示例性的实施方案中,本公开提供了用于促进毛发生长的药物组合物,其包含弯曲乳杆菌WIKIM55或其培养物作为活性成分。
当本公开的组合物用作药物组合物时,除了活性成分之外,可以使用药学上合适且生理上可接受的佐剂来制备本公开的药物组合物。可以使用赋形剂、崩解剂、甜味剂、黏合剂、包衣剂、溶胀剂、润滑剂、助流剂、调味剂等作为佐剂。
具体地,除了上述用于活性成分外,可以通过添加一种或多于一种药学上可接受的载剂来配制药物组合物,以施用该药物组合物。
药物组合物的制剂形式可以是颗粒剂、散剂、片剂、包衣片剂、胶囊剂、栓剂、溶液、糖浆剂、汁剂、混悬剂、乳剂、滴剂、注射液等。例如,为了配制成片剂或胶囊剂的形式,可以将活性成分与口服、无毒、药学上可接受的惰性载剂例如乙醇、甘油、水等组合。此外,如果希望或需要的话,混合物中还可以包含合适的黏合剂、润滑剂、崩解剂和着色剂。合适的黏合剂包括但不限于淀粉、明胶、天然糖例如葡萄糖或β-乳糖、玉米甜味剂、天然或合成树胶例如阿拉伯胶、黄芪胶或油酸钠、硬脂酸钠、硬脂酸镁、苯甲酸钠、乙酸钠、氯化钠等。崩解剂包括但不限于淀粉、甲基纤维素、琼脂、膨润土、黄原胶等。
如在配制成液体溶液的组合物中的可接受的药物载剂中,可以混合经灭菌且适用于活体的生理盐水、无菌水、林格氏溶液、缓冲盐水、白蛋白注射液、葡萄糖溶液、麦芽糖糊精溶液、甘油和乙醇中的一种或多于一种成分。如果需要,可以添加其他常规添加剂,例如抗氧化剂、缓冲剂、抑菌剂等。另外,可以另外添加稀释剂、分散剂、表面活性剂、黏合剂或润滑剂以配制成可注射制剂例如水溶液、混悬剂、乳剂、丸剂、胶囊剂、颗粒剂、或片剂等。
此外,可以使用Remington's Pharmaceutical Science,Mack PublishingCompany,Easton PA中公开的方法根据相应的疾病或成分进行配制。
另外,本公开提供了促进毛发生长的方法,该方法包括将治疗有效量的弯曲乳杆菌WIKIM55或其培养物施用于有需要的对象。
本文使用的术语“对象”是指作为治疗、观察或实验对象的哺乳动物。具体来说,对象指人类。
此外,本文所用的术语“治疗有效量”是指引起组织系统、动物或人类中的生物或医学反应的活性成分或药物组合物的量,其由研究者、兽医、医生或临床医师确定。其包括诱导减轻所治疗疾病或病症的症状的量。对于本领域技术人员显而易见的是,本公开的活性成分的治疗有效量和给药次数将根据所需效果而改变。因此,本领域技术人员可以容易地确定最佳的给药剂量,并且可以根据多种因素进行调整,该因素包括疾病的类型、疾病的严重程度、活性成分的含量和组合物中包含的其他成分、制剂的类型、患者的年龄、体重、总体健康状况、性别和饮食、给药时间、给药途径、组合物的排泄率、治疗周期和共用药物。具体地,在本公开的治疗方法中,弯曲乳杆菌WIKIM55或其培养物可以按照每天一次或几次对于成年人以0.01mg/kg至200mg/kg的剂量施用。
在本发明的治疗方法中,包含弯曲乳杆菌WIKIM55或其培养物作为活性成分的本公开的组合物可以按照常规方法通过经口、经直肠、静脉内、动脉内、腹膜内、肌内、胸骨内、透皮、局部、眼内或皮内途径施用。
在另一个示例性的实施方案中,本公开提供了用于促进毛发生长的食品组合物,其包含弯曲乳杆菌WIKIM55或其培养物作为活性成分。食物组合物可以是功能性食品、饮料、营养棒等的形式。
在本公开中,包含菌株作为活性成分的食品组合物可以包括饮料例如发酵乳等。因此,本公开提供了用于发酵的乳酸菌发酵剂,其包含弯曲乳杆菌WIKIM55或其培养物。
根据本公开的食品组合物可以按照与药物组合物相同的方式配制,并且可以用作功能性食品或添加到各种食品中。可以向其中添加本公开的组合物的食品包括例如饮料、酒精饮料、甜食、营养棒(diet bar)、乳制品、肉、巧克力、比萨、方便面、口香糖、冰淇淋、维生素复合物、健康补充剂等。
例如,如果将本公开的食品组合物制备成饮料例如饮品(drink),则除了弯曲乳杆菌WIKIM55或其培养物之外,其还可以包含柠檬酸、果糖糖浆、糖、葡萄糖、乙酸、苹果酸、果汁、各种植物提取物等。
另外,本公开可以提供包含弯曲乳杆菌WIKIM55或其培养物的功能性食品。功能性食品是指通过将弯曲乳杆菌WIKIM55或其培养物添加到例如饮品、茶、口香糖、甜食等食品原料中而制成的食品,或者通过胶囊化、粉碎、悬浮等方法制成的食品,如果摄入它们,将带来特定的健康益处,但是与普通药物不同,它们具有长期使用不会产生副作用的优点。因此,如此获得的本公开的功能性食品非常有用,因为其可以常规地摄入。在这种保健食品中,弯曲乳杆菌WIKIM55或其培养物的添加量不能统一限定,因为其可能根据保健食品的类型而变化。该量可以在不损害食品的原始味道的范围内,并且相对于所添加到的食品,其可以为0.01重量%至50重量%,特别是0.1重量%至20重量%。对于颗粒、片剂或胶囊形式的食品,其添加量通常可以为0.1重量%至100重量%,特别是0.5重量%至80重量%。在一个示例性实施方案中,本公开的功能性食品可以是饮品的形式。
根据本公开的组合物可以用作饲料添加剂或饲料。
当用作饲料添加剂时,可以将组合物制成20%至90%的高浓度溶液、或粉末或颗粒。饲料添加剂还可以包含有机酸例如柠檬酸、富马酸、己二酸、乳酸、苹果酸等,磷酸盐例如磷酸钠、磷酸钾、酸性焦磷酸盐、多磷酸盐等,或天然抗氧化剂例如多酚、儿茶素、α-生育酚、迷迭香提取物、维生素C、绿茶提取物、甘草根提取物、壳聚糖、单宁酸、植酸等中的一种或多于一种。当用作饲料时,可以将组合物配制成普通饲料,并且组合物还可以包含常用的饲料成分。
饲料添加剂和饲料还可包含谷物,例如磨碎或压碎的小麦、燕麦、大麦、玉米和大米;植物蛋白饲料,例如以油菜、大豆和向日葵为主要成分的饲料;动物蛋白饲料,例如血粉、金属粉、骨粉和鱼粉;糖或奶,例如由各种奶粉和乳清粉组成的干燥成分。另外,还可以包含营养补充剂、消化吸收促进剂、生长促进剂等。
饲料添加剂可以单独施用于动物或在可食用载剂中与其他饲料添加剂组合施用于动物。此外,饲料添加剂可以作为追肥直接混入动物饲料中,或者可以与饲料分开作为经口剂型容易地施用于动物。当饲料添加剂与动物饲料分开施用时,可以将其与药学上可接受的可食用载剂组合制成速释或缓释制剂,这是本领域众所周知的。可食用载剂可以是固体或液体,例如玉米淀粉、乳糖、蔗糖、大豆片、花生油、橄榄油、芝麻油和丙二醇。如果使用固体载剂,则饲料添加剂可以是片剂、胶囊剂、粉剂、锭剂、糖衣片剂或追肥的形式。如果使用液体载剂,则饲料添加剂可以是软明胶胶囊剂、糖浆剂、混悬剂、乳剂或溶液的形式。
此外,饲料添加剂和饲料可以包含佐剂,例如防腐剂、稳定剂、润湿剂或乳化剂、增溶剂等。饲料添加剂可以通过喷雾或混合添加到动物饲料中。
本公开的饲料或饲料添加剂可以提供给许多动物,包括哺乳动物、家禽和鱼。
哺乳动物可以包括猪、牛、绵羊、山羊、实验啮齿动物和宠物(例如狗和猫)。家禽可以包括但不限于鸡、火鸡、鸭、鹅、野鸡、鹌鹑等,鱼可以包括但不限于鳟鱼等。
在一个示例性的实施方案中,饲料或饲料添加剂可用于促进宠物的毛发生长。宠物包括但不限于狗、猫、大鼠、兔等。
此外,当组合物用作化妆品组合物时,所述化妆品组合物可以配制成用于皮肤或头皮清洁剂的外用组合物,但不特别限于此。用于皮肤的外用组合物可以是例如柔肤水、滋养化妆水、按摩霜、滋养霜、面膜或凝胶,并且可以是用于透皮施用的制剂,例如化妆水、软膏、凝胶、乳霜、贴剂或喷雾剂。头皮清洁剂可以配制成例如喷发剂、生发剂、护发素、护发精华、护发乳、滋养发乳、洗发香波、洗发水、头发护理剂、发膏、发用滋养霜、发用保湿霜、发用按摩霜、发蜡、头发气雾剂、发膜、发用营养膜、洗发皂、洗发泡沫、染发剂、卷发剂、头发漂白剂、发胶、发釉或发用摩丝,但不限于此。在各个制剂的外用组合物中,根据外用组合物的剂型或用途,可以添加本领域技术人员容易选择的除弯曲乳杆菌WIKIM55菌株以外的成分。
具体地,本公开的化妆品组合物包含化妆品可接受的介质或基质。其可以作为适用于局部应用的所有可能的制剂形式提供,例如溶液、凝胶、固体或无水膏剂、通过将油相分散在水相中而获得的乳液、混悬液、微乳液、微胶囊、微颗粒、离子(脂质体)和/或非离子囊泡分散液、乳霜、皮肤化妆水、粉末、软膏、喷雾剂或遮瑕膏。此外,也可以将其制成还包含压缩推进剂的泡沫或气溶胶组合物。另外,通过包含皮肤病学上可接受的介质或基质,可以将本公开的化妆品组合物制备成在皮肤病学领域中常用的可局部或全身应用的佐剂,并且这些组合物可以根据现有技术中常用的方法制备。
另外,本公开的化妆品组合物可以包含皮肤病学领域常用的佐剂,例如脂肪物质、有机溶剂、增溶剂、增稠剂、胶凝剂、软化剂、抗氧化剂、助悬剂、稳定剂、发泡剂、添味剂、表面活性剂、水、离子或非离子乳化剂、填充剂、金属离子螯合剂、螯合剂、防腐剂、维生素、阻滞剂、保湿剂、精油、染料、颜料、亲水或亲脂性活化剂、脂质囊泡或化妆品中常用的任何其他成分。这些佐剂以化妆品领域中常用的量掺入。
基于单一剂量,包含在根据本公开的组合物中的弯曲乳杆菌WIKIM55菌株的量可以为约106CFU/g至1012CFU/g,例如107CFU/g至1011CFU/g或108CFU/g至1010CFU/g。具体地,该菌株可以作为活细菌施用,并且可以在摄入之前被杀死或弱化。此外,当通过使用培养上清液等制备组合物时,可以另外通过热处理进行灭菌过程。实现最小效果所需的菌株的量及其日剂量可以根据患者身体或健康状况而变化。通常,日剂量可以为约106CFU/g至1012CFU/g,例如107CFU/g至1011CFU/g或108CFU/g至1010CFU/g。
通过下面详细描述的实施例,优点和特征以及实现它们的方法将变得显而易见。然而,本公开不限于以下阐述的实施例,而是可以以许多不同的形式来体现。提供实施例以使得本公开的公开是完整的,并且提供实施例以将本发明的范围充分告知本公开所属领域的普通技术人员。本公开的范围将仅由所附权利要求限定。
有益效果
因为根据本公开的弯曲乳杆菌WIKIM55表现出促进毛发生长的活性,所以其可以作为益生菌用于多种用途,例如人或动物中的肠道调节、增强免疫力、促进毛发生长等。此外,它可以有效地用作发酵的发酵剂。
附图说明
图1示出了用本公开的菌株饲喂30天的小鼠的毛发生长模式的肉眼观察结果。
图2示出了用本公开的菌株饲喂的小鼠的皮肤组织以及测量皮肤厚度的结果。
图3示出了用本公开的菌株饲喂的小鼠的皮下组织中毛囊数量的测量结果。
图4示出了观察用本公开的菌株饲喂20周的小鼠的毛发生长模式并鉴定毛发生长初期的结果。
图5示出了通过FACS分析确认用本公开的菌株饲喂的小鼠的毛囊干细胞分布的结果。
最佳实施方式
以下会通过实施例详细地描述本公开。然而,以下实施例仅出于说明目的,并且本公开的范围不受实施例限制。
[实施例]
实施例1:菌株的分离与鉴定
收集通过在MRS培养基中涂抹泡菜粗制液体提取物而获得的细菌单菌落,并在MRS液体培养基中培养。使用QIAamp DNA微量试剂盒(QIAgen,德国)提取DNA。使用1%琼脂糖凝胶确认提取的DNA。以提取的基因组DNA为模板进行PCR以扩增16S rDNA基因。PCR条件是30个以下步骤的循环:在95℃下变性1分钟,在45℃下退火1分钟和在72℃延伸1分钟30秒。获得的PCR产物的序列由Macrogen(韩国首尔)分析。通过美国国家生物技术信息中心(NCBI,www.ncbi.nlm.nih.gov)的基本局部比对搜索工具(BLAST)搜索引擎对16S rDNA序列进行相似性分析来进行细菌鉴定。
作为用于鉴定微生物的16S rDNA碱基序列分析的结果,发现在该实施例中分离的菌株具有SEQ ID NO:1的核酸序列。
本公开的微生物被命名为弯曲乳杆菌WIKIM55,并于2017年4月7日保藏在韩国微生物保藏中心(保藏号:KCCM12011P)。
SEQ ID NO:1.弯曲乳杆菌WIKIM55的16S rDNA序列
实施例2:确认弯曲乳杆菌WIKIM55促进毛发生长的作用
将实施例1中分离的弯曲乳杆菌WIKIM55菌株在MRS培养基中于30℃培养24小时,然后将培养的细菌在8000rpm下离心5分钟,然后用PBS洗涤以除去残留的培养基成分。然后,使用PBS将细菌的数量定量至1×1010CFU/mL,并且使用探头每周五次将0.2mL(2×109CFU)经口施用至测试动物。将无菌PBS施用于阴性对照组和阳性对照组。
1)感官评估
将毛发生长终期之前的6周龄小鼠(C57/BL6)在繁殖室中放置一周,然后使用动物用脱毛器去除背部的毛发。然后,经口施用弯曲乳杆菌WIKIM55 30天后,通过临床视觉评估来评估毛发生长模式。
结果如图1所示,与饲喂PBS的对照组(PBS)相比,饲喂弯曲乳杆菌WIKIM55菌株的组(WIKIM55)表现出更快的毛发生长。
2)皮肤厚度的测量
毛发在整个毛发周期中重复生长和脱落。众所周知,在毛发生长的生长初期,皮肤层变厚并且毛囊主要存在于皮肤的皮下层。因此,在去除小鼠毛发后30天检查皮肤组织和皮肤厚度(图2)。
结果,PBS处理组的皮肤厚度为393.28μm,而用弯曲乳杆菌WIKIM55饲喂的组的皮肤厚度为670.95μm。还证实大多数毛囊存在于皮下层中。由此证实,用弯曲乳杆菌WIKIM55饲喂的组的大多数毛囊处于生长初期。
3)皮下组织中毛囊数量的测量
在去除小鼠毛发后30天,测量皮下组织中毛囊的数量。
结果如图3所示,用弯曲乳杆菌WIKIM55饲喂的组的毛囊数量显著大于PBS处理的组。
实施例3:确认弯曲乳杆菌WIKIM55促进毛发生长的作用
将实施例1中分离的弯曲乳杆菌WIKIM55菌株在MRS培养基中于30℃培养24小时,然后将培养的细菌在8000rpm下离心5分钟,然后用PBS洗涤以除去残留的培养基成分。然后,使用PBS将细菌的数量定量至1×1010CFU/mL,并且使用探头每周五次将0.2mL(2×109CFU)经口施用至测试动物。将无菌PBS施用于阴性对照组和阳性对照组。
1)感官评估
将毛发生长终期之前的6周龄小鼠(C57/BL6)在繁殖室中放置一周,然后使用动物用脱毛器去除背部的毛发。经口施用弯曲乳杆菌WIKIM55 20周后,通过临床视觉评估来评估毛发生长模式。
如图4所示,饲喂PBS的对照组(PBS)在休止期约7周后处于毛发生长初期,而饲喂弯曲乳杆菌WIKIM55菌株的组(WIKIM55)在休止期约2周至3周后转变为毛发生长初期,从而导致更快的毛发生长。
2)确认毛囊干细胞的分布
为了比较对照组(PBS)和用弯曲乳杆菌WIKIM55饲喂20周(WIKIM55)的小鼠的毛囊干细胞的分布,切下背部的皮肤组织。从切下的组织中去除皮下脂肪,然后用磷酸盐缓冲盐水(PBS)洗涤后,将组织置于RPMI1640培养基中,该培养基包含4mg/mL的胶原蛋白酶IV、2mg/mL的透明质酸酶和100U/mL的DNA酶I,并且使用gentleMACS分离器(Miltenyibiotec,德国)从皮肤组织中分离毛囊干细胞。进行FACS(荧光激活细胞分选)分析以研究毛囊干细胞的分布。验证的抗原显示出CD45-CD34+CD49f+的免疫学特征。用含有2%FBS的PBS溶液洗涤从皮肤组织分离的1×105个细胞,并使其在室温下与针对相应抗原的抗体反应。通过流式细胞术确认抗原的表达。
从图5中可以看出,证实了与对照组(PBS,未处理)相比,在用WIKIM55饲喂的组的皮肤组织中分布有更多的表现出CD34+CD49f+免疫学特征的毛囊干细胞。由于毛囊干细胞参与了新毛发的生长,因此认为WIKIM55的摄入将通过产生毛囊干细胞来促进毛发的生长。
序列表
<110> 韩国食品研究院
<120> 具有促进毛发生长活性的弯曲乳芽孢杆菌WIKIM55和包含其的组合物
<130> X18R10C0070
<150> KR 10-2017-0068238
<151> 2017-06-01
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1449
<212> DNA
<213> 弯曲乳芽孢杆菌WIKIM55(Lactobacillus curvatus WIKIM55)
<400> 1
acatgcaagt cgaacgcact ctcgttagat tgaagaagct tgcttctgat tgataacatt 60
tgagtgagtg gcggacgggt gagtaacacg tgggtaacct gccctaaagt gggggataac 120
atttggaaac agatgctaat accgcataaa acctagcacc gcatggtgca aggttgaaag 180
atggtttcgg ctatcacttt aggatggacc cgcggtgcat tagttagttg gtgaggtaaa 240
ggctcaccaa gaccgtgatg catagccgac ctgagagggt aatcggccac actgggactg 300
agacacggcc cagactccta cgggaggcag cagtagggaa tcttccacaa tggacgaaag 360
tctgatggag caacgccgcg tgagtgaaga aggttttcgg atcgtaaaac tctgttgttg 420
gagaagaacg tatttgatag taactgatca ggtagtgacg gtatccaacc agaaagccac 480
ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt ccggatttat 540
tgggcgtaaa gcgagcgcag gcggtttctt aagtctgatg tgaaagcctt cggctcaacc 600
gaagaagtgc atcggaaact gggaaacttg agtgcagaag aggacagtgg aactccatgt 660
gtagcggtga aatgcgtaga tatatggaag aacaccagtg gcgaaggcgg ctgtctggtc 720
tgtaactgac gctgaggctc gaaagcatgg gtagcaaaca ggattagata ccctggtagt 780
ccatgccgta aacgatgagt gctaggtgtt ggagggtttc cgcccttcag tgccgcagct 840
aacgcattaa gcactccgcc tggggagtac gaccgcaagg ttgaaactca aaggaattga 900
cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagcaacgcg aagaacctta 960
ccaggtcttg acatcctttg accactctag agatagagct ttcccttcgg ggacaaagtg 1020
acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080
gagcgcaacc cttattacta gttgccagca tttagttggg cactctagtg agactgccgg 1140
tgacaaaccg gaggaaggtg gggacgacgt caaatcatca tgccccttat gacctgggct 1200
acacacgtgc tacaatggat ggtacaacga gtcgcgagac cgcgaggttt agctaatctc 1260
ttaaaaccat tctcagttcg gattgtaggc tgcaactcgc ctacatgaag ccggaatcgc 1320
tagtaatcgc ggatcagcat gccgcggtga atacgttccc gggccttgta cacaccgccc 1380
gtcacaccat gaagagtttg taacacccaa agccgggtga ggtaaccttc gggagccagc 1440
cgtctaagg 1449
Claims (7)
1.一种保藏号为KCCM12011P的弯曲乳杆菌(Lactobacillus curvatus)WIKIM55。
2.一种用于促进毛发生长的药物组合物,其包含保藏号为KCCM12011P的弯曲乳杆菌WIKIM55。
3.根据权利要求2所述的药物组合物,其中组合物是经口施用的药物组合物。
4.一种用于增强免疫力和肠道调节的组合物,其包含保藏号为KCCM12011P的弯曲乳杆菌WIKIM55。
5.一种用于发酵的乳酸菌发酵剂,其包含保藏号为KCCM12011P的弯曲乳杆菌WIKIM55。
6.一种用于促进毛发生长的化妆品组合物,其包含保藏号为KCCM12011P的弯曲乳杆菌WIKIM55。
7.保藏号为KCCM12011P的弯曲乳杆菌WIKIM55菌株用于促进毛发生长的用途。
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AU2018276725A1 (en) | 2019-12-19 |
EP3636744B1 (en) | 2024-02-21 |
WO2018221956A2 (ko) | 2018-12-06 |
EP3636744C0 (en) | 2024-02-21 |
JP6861853B2 (ja) | 2021-04-21 |
JP2020521475A (ja) | 2020-07-27 |
AU2018276725B2 (en) | 2021-01-07 |
US20200178565A1 (en) | 2020-06-11 |
CN110741074A (zh) | 2020-01-31 |
CA3065083A1 (en) | 2018-12-06 |
EP3636744A2 (en) | 2020-04-15 |
CA3065083C (en) | 2022-09-13 |
US11559064B2 (en) | 2023-01-24 |
KR101801764B1 (ko) | 2017-11-27 |
EP3636744A4 (en) | 2021-03-03 |
WO2018221956A3 (ko) | 2019-03-28 |
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