CN112806576B - 抑制阴道炎病原菌的组合物、阴道清洁组合物以及其用途 - Google Patents
抑制阴道炎病原菌的组合物、阴道清洁组合物以及其用途 Download PDFInfo
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- CN112806576B CN112806576B CN201911038378.XA CN201911038378A CN112806576B CN 112806576 B CN112806576 B CN 112806576B CN 201911038378 A CN201911038378 A CN 201911038378A CN 112806576 B CN112806576 B CN 112806576B
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Abstract
本发明提供了一种抑制阴道炎病原菌的组合物、阴道清洁组合物以及其用途。具体地,本发明提供一种用以抑制阴道炎病原菌的组合物,其包含乳酸菌发酵物,其是于包含牛奶、奶粉及酪蛋白至少其中之一的培养基中以唾液乳酸杆菌AP‑32菌株、鼠李糖乳酸杆菌F‑1菌株、鼠李糖乳酸杆菌GL‑165菌株、动物双歧杆菌乳亚种CP‑9菌株、两歧双歧杆菌Bf‑688菌株、短双歧杆菌Bv‑889菌株或以上乳酸菌菌株的组合与嗜热链球菌共同进行发酵而得。上述乳酸菌菌株发酵物可抑制阴道炎病原菌生长,并以食品组合物、医药及阴道清洁组合物的形式存在。
Description
技术领域
本发明是有关一种组合物、阴道清洁组合物以及其用途,特别是关于一种用以抑制阴道炎病原菌的组合物、阴道清洁组合物以及其用途。
背景技术
有关于益生菌对于人体健康的研究十分广泛,一般来说,对于人的身体健康有特殊功效的菌株称为功能性益生菌(Guidelines for the evaluation of probiotics infood;Report of joint FAO/WHO working group on drafting guidelines for theevaluation of probiotics in food;London Ontario,Canada April 30and May 1,2002:1-7)。常见的功能性益生菌的特殊功效包含调整肠胃道功能、调节免疫能力、甚至是抗发炎反应等。需注意的是,上述功能性益生菌的特殊功效在于菌株(strain)的特异性而非菌种(species),亦即,即使是相同的菌种,若为不同的菌株,仍可能具有不同的功效,甚至具有相反的功效。
一些研究指出,现代妇女约有超过50%的机率发生尿道或阴道感染。在门诊就诊的妇女中,常见诸如排尿困难、瘙痒和分泌物等下生殖道症状。最常见的原因是外阴道感染,育龄妇女的外阴道感染有三个常见原因:念珠菌病、细菌性阴道疾病,以及滴虫病,其中前两者都是由微生物所造成。阴道与外界相通,又靠近有很多潜在病原菌的肛门,因此会有很多机会遭到病原菌侵入,所以人体有一套防御系统以避免病原菌过度增生。
阴道的防御系统主要基础是pH值低于4.5的酸性环境。酸性环境不利于潜在病原菌的增生,而对嗜酸性的杆菌则是理想的环境,因此可以使嗜酸性杆菌持续产酸,维持一个健康的状态。然而,若是酸性细菌活性降低,例如使用广效抗生素等原因,则会使阴道酸碱度上升,导致病原菌增生并造成感染。
有鉴于此,开发具有抑制阴道带病原菌功效的产品是目前需努力的目标。
发明内容
本发明提供一种用以抑制阴道炎病原菌的组合物,其具有于包含牛奶、奶粉以及酪蛋白至少其中之一的培养基中以乳酸菌菌株与嗜热链球菌共同进行发酵所得的发酵物。上述乳酸菌菌株发酵物可抑制阴道炎病原菌生长,且以食品组合物、医药及阴道清洁组合物的形式存在。
本发明一实施例的用以抑制阴道炎病原菌的组合物包含乳酸菌发酵物以及赋形剂、稀释剂或载体。乳酸菌发酵物是以包含牛奶、奶粉以及酪蛋白至少其中之一的培养基,并以经分离的乳酸菌菌株与嗜热链球菌共同进行发酵而获得,其中乳酸菌菌株包含保藏编号为CCTCC NO:M2011127的唾液乳酸杆菌(Lactobacillus salivariussubsp.salicinius)AP-32菌株(保藏日期:2011年4月10日;保藏单位:中国典型培养物保藏中心(CCTCC);保藏单位地址:中国武汉武汉大学,邮编430072;分类命名:唾液乳酸杆菌(Lactobacillus salivarius subsp.salicinius))、保藏编号为CCTCC NO:M2011124的鼠李糖乳酸杆菌(Lactobacillus rhamnosus)F-1菌株(保藏日期:2011年4月10日;保藏单位:中国典型培养物保藏中心(CCTCC);保藏单位地址:中国武汉武汉大学,邮编430072;分类命名:鼠李糖乳酸杆菌(Lactobacillus rhamnosus))、保藏编号为CCTCC NO:M2014591的鼠李糖乳酸杆菌(Lactobacillus rhamnosus)GL-165菌株(保藏日期:2014年11月24日;保藏单位:中国典型培养物保藏中心(CCTCC);保藏单位地址:中国武汉武汉大学,邮编430072;分类命名:鼠李糖乳酸杆菌(Lactobacillus rhamnosus))、保藏编号为CCTCC NO:M2014588的动物双歧杆菌乳亚种(Bifidobacterium animalis subsp.lactis)CP-9菌株(保藏日期:2014年11月24日;保藏单位:中国典型培养物保藏中心(CCTCC);保藏单位地址:中国武汉武汉大学,邮编430072;分类命名:动物双歧杆菌乳亚种(Bifidobacterium animalissubsp.lactis))、保藏编号为CGMCC No.17953的两歧双歧杆菌(Bifidobacteriumbifidum)Bf-688菌株(保藏日期:2019年06月18日;保藏单位:中国微生物菌种保藏管理委员会普通微生物中心(CGMCC);保藏单位地址:北京市朝阳区北辰西路1号院3号中国科学院微生物研究所,邮编100101;分类命名:两歧双歧杆菌(Bifidobacterium bifidum))、保藏编号为CGMCC No.16145的短双歧杆菌(Bifidobacterium breve)Bv-889菌株(保藏日期:2018年07月23日;保藏单位:中国微生物菌种保藏管理委员会普通微生物中心(CGMCC);保藏单位地址:北京市朝阳区北辰西路1号院3号中国科学院微生物研究所,邮编100101;分类命名:短双歧杆菌(Bifidobacterium breve))或以上乳酸菌菌株的组合。上述乳酸菌菌株分别保藏于中国典型培养物保藏中心以及中国微生物菌种保藏管理委员会普通微生物中心。
本发明另一实施例的阴道清洁组合物包含乳酸菌发酵物以及生理上可接受的赋形剂、稀释剂或载体。乳酸菌发酵物是以包含牛奶、奶粉以及酪蛋白至少其中之一的培养基,并以经分离的乳酸菌菌株与嗜热链球菌共同进行发酵而获得,其中该乳酸菌菌株包含保藏编号为CCTCC NO:M2011127的唾液乳酸杆菌AP-32菌株、保藏编号为CCTCC NO:M2011124的鼠李糖乳酸杆菌F-1菌株、保藏编号为CCTCC NO:M2014591的鼠李糖乳酸杆菌GL-165菌株、保藏编号为CCTCC NO:M2014588的动物双歧杆菌乳亚种CP-9菌株、保藏编号为CGMCC No.17953的两歧双歧杆菌Bf-688菌株、保藏编号为CGMCC No.16145的短双歧杆菌Bv-889菌株或以上乳酸菌菌株的组合。上述乳酸菌菌株分别保藏于中国典型培养物保藏中心以及中国微生物菌种保藏管理委员会普通微生物中心。
本发明另一实施例的以包含乳酸菌发酵物的组合物作为抑制阴道炎病原菌的用途,其中包含乳酸菌发酵物的组合物包含乳酸菌发酵物以及赋形剂、稀释剂或载体。乳酸菌发酵物是以包含牛奶、奶粉以及酪蛋白至少其中之一的培养基,并以经分离的乳酸菌菌株与嗜热链球菌共同进行发酵而获得,其中该乳酸菌菌株包含保藏编号为CCTCC NO:M2011127的唾液乳酸杆菌AP-32菌株、保藏编号为CCTCC NO:M2011124的鼠李糖乳酸杆菌F-1菌株、保藏编号为CCTCC NO:M2014591的鼠李糖乳酸杆菌GL-165菌株、保藏编号为CCTCCNO:M2014588的动物双歧杆菌乳亚种CP-9菌株、保藏编号为CGMCC No.17953的两歧双歧杆菌Bf-688菌株、保藏编号为CGMCC No.16145的短双歧杆菌Bv-889菌株或以上乳酸菌菌株的组合。上述乳酸菌菌株分别保藏于中国典型培养物保藏中心以及中国微生物菌种保藏管理委员会普通微生物中心。
以下藉由具体实施例配合所附的图式详加说明,当更容易了解本发明的目的、技术内容、特点及其所达成的功效。
附图说明
图1显示本发明的各别乳酸菌代谢物抑制阴道炎病原菌的结果。
图2显示本发明的各别乳酸菌代谢物抑制阴道炎病原菌的结果。
图3显示本发明的混合乳酸菌菌株的发酵物抑制阴道炎病原菌的结果。
图4显示一直方图,显示本发明的乳酸菌菌株的混合发酵物抑制阴道炎病原菌的结果。
图5显示一直方图,显示本发明的不同浓度的发酵物凝胶抑制阴道炎病原菌的结果。
用于专利程序的微生物保藏:
(一)AP-32菌株
保藏编号:CCTCC NO:M2011127;
保藏日期:2011年4月10日;
保藏单位:中国典型培养物保藏中心(CCTCC);
保藏单位地址:中国武汉武汉大学,邮编430072;
分类命名:唾液乳酸杆菌(Lactobacillus salivarius subsp.salicinius);
(二)F-1菌株
保藏编号:CCTCC NO:M2011124;
保藏日期:2011年4月10日;
保藏单位:中国典型培养物保藏中心(CCTCC);
保藏单位地址:中国武汉武汉大学,邮编430072;
分类命名:鼠李糖乳酸杆菌(Lactobacillus rhamnosus);
(三)GL-165菌株
保藏编号:CCTCC NO:M2014591;
保藏日期:2014年11月24日;
保藏单位:中国典型培养物保藏中心(CCTCC);
保藏单位地址:中国武汉武汉大学,邮编430072;
分类命名:鼠李糖乳酸杆菌(Lactobacillus rhamnosus);
(四)CP-9菌株
保藏编号:CCTCC NO:M2014588;
保藏日期:2014年11月24日;
保藏单位:中国典型培养物保藏中心(CCTCC);
保藏单位地址:中国武汉武汉大学,邮编430072;
分类命名:动物双歧杆菌乳亚种(Bifidobacterium animalis subsp.lactis);
(五)Bf-688菌株
保藏编号:CGMCC No.17953;
保藏日期:2019年06月18日;
保藏单位:中国微生物菌种保藏管理委员会普通微生物中心(CGMCC);
保藏单位地址:北京市朝阳区北辰西路1号院3号中国科学院微生物研究所,邮编100101;
分类命名:两歧双歧杆菌(Bifidobacterium bifidum);
(六)Bv-889菌株
保藏编号:CGMCC No.16145;
保藏日期:2018年07月23日;
保藏单位:中国微生物菌种保藏管理委员会普通微生物中心(CGMCC);
保藏单位地址:北京市朝阳区北辰西路1号院3号中国科学院微生物研究所,邮编100101;
分类命名:短双歧杆菌(Bifidobacterium breve)。
具体实施方式
以下将详述本发明的各实施例,并配合附图作为例示。除了这些详细说明之外,本发明亦可广泛地施行于其它的实施例中,任何所述实施例的轻易替代、修改、等效变化都包含在本发明的范围内,并以请求保护范围为准。在说明书的描述中,为了使读者对本发明有较完整的了解,提供了许多特定细节;然而,本发明可能在省略部分或全部特定细节的前提下,仍可实施。此外,众所周知的步骤或组件并未描述于细节中,以避免对本发明形成不必要的限制。附图中相同或类似的组件将以相同或类似符号来表示。特别注意的是,附图仅为示意之用,并非代表组件实际的尺寸或数量,有些细节可能未完全绘出,以求附图的简洁。
本发明所述的乳酸菌菌株的冷冻干燥培养物已分别保藏于中国典型培养物保藏中心(CCTCC,地址为中国武汉武汉大学,430072)以及中国微生物菌种保藏管理委员会普通微生物中心(CGMCC,地址为北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所)。保藏的详细资料如表1所示:
表1乳酸菌菌株的保藏资料
如表1所列已保藏的乳酸菌菌株的发酵物被发现具有抑制阴道炎病原菌的活性效果。因此,表1所列已保藏的乳酸菌菌株的发酵物可作为抑制阴道炎病原菌的用途。
于一实施例中,本发明的用以抑制阴道炎病原菌的组合物包含乳酸菌发酵物以及赋形剂、稀释剂或载体。乳酸菌发酵物是以包含牛奶、奶粉、酪蛋白或以上的组合为基底的培养基,并以经分离的乳酸菌菌株与嗜热链球菌共同进行发酵而获得。乳酸菌菌株可为保藏编号为CCTCC NO:M2011127的唾液乳酸杆菌(Lactobacillus salivariussubsp.salicinius)AP-32菌株、保藏编号为CCTCC NO:M2011124的鼠李糖乳酸杆菌(Lactobacillus rhamnosus)F-1菌株、保藏编号为CCTCC NO:M2014591的鼠李糖乳酸杆菌(Lactobacillus rhamnosus)GL-165菌株、保藏编号为CCTCC NO:M2014588的动物双歧杆菌乳亚种(Bifidobacterium animalis subsp.lactis)CP-9菌株、保藏编号为CGMCCNo.17953的两歧双歧杆菌(Bifidobacterium bifidum)Bf-688菌株、保藏编号为CGMCCNo.16145的短双歧杆菌(Bifidobacterium breve)Bv-889菌株或以上乳酸菌菌株的组合,上述乳酸菌菌株分别保藏于中国典型培养物保藏中心以及中国微生物菌种保藏管理委员会普通微生物中心。
于一实施例中,赋形剂、稀释剂或载体可为生理上可接受的赋形剂、稀释剂或载体,使本发明的组合物作为食品组合物或阴道清洁组合物使用。于一实施例中,赋形剂、稀释剂或载体可为医药上可接受的赋形剂、稀释剂或载体,使本发明的组合物作为一医药组合物使用。
于食品组合物的实施例中,生理上可接受的赋形剂、稀释剂或载体可为一食品。举例而言,食品可包含但不限于乳制饮品、茶、咖啡、功能性饮品或以上的组合,其中乳制饮品可包含发酵乳、优格、奶酪或乳制饮品乳粉等。
于医药组合物的实施例中,医药组合物可为口服剂型或外用剂型。举例而言,口服剂型可为锭剂、胶囊、溶液剂或粉剂等;外用剂型可为粉剂、乳膏、喷雾剂、凝胶、溶液剂、散剂或霜剂等。
于阴道清洁组合物的实施例中,阴道清洁组合物可为锭剂、胶囊、粉剂、乳膏、喷雾剂、凝胶、溶液剂、散剂、霜剂或洗剂。
于一实施例中,本发明的乳酸菌菌株所发酵产生的发酵物可包含去活性菌株或去除菌体的发酵液或其干燥粉末。举例而言,发酵液可为发酵上清液或乳清发酵液等。于一实施例中,本发明的用于抑制阴道炎病原菌的组合物中乳酸菌发酵物的粉剂含量为0.4%以上;或者,乳酸菌发酵物的溶液含量为2%以上。
于一实施例中,本发明的用以抑制阴道炎病原菌的组合物还包含消炎剂或防腐剂。举例而言,消炎剂或防腐剂可为第一复合物、第二复合物、第三复合物、薄荷醇油、茶树精油、芦荟萃取物、左手香萃取物、纳米银或以上的组合。第一复合物包含二丙二醇(dipropylene glycol)、羟基苯乙酮(hydroxyacetophenone)、辛基乙二醇(caprylylglycol)以及甘草酸二钾(dipotassium glycyrrhizinate),业界称为activonol-M。第二复合物包含野大豆籽萃取物(Glycine Soja Seed Extract)、鱼腥草萃取物(HouttuyniaCordata Extract)、黄芩根萃取物(Scutellaria Baicalensis Root Extract)、印度楝叶萃取物(Melia Azadirachta Leaf Extract)、中国地黄根萃取物(Rehmannia ChinensisRoot Extract)、白柳树皮萃取物(Salix Alba Bark Extract)、黄蘖树皮萃取物(Phellodendron Amurense Bark Extract)、o-伞花烃-5-醇(o-Cymen-5-Ol)、乳酸杆菌、梨汁发酵滤液(Pear Juice Ferment Filtrate)、甘油辛酸酯(Glyceryl Caprylate)、乙基己基甘油(Ethylhexylglycerin)、PEG-60氢化蓖麻油(PEG-60Hydrogenated Castor Oil)、羟基乙酸(Glycolic Acid)、丁二醇(Butylene Glycol)及水,业界称为acnebusters。第三复合物包含秦椒果萃取物(Zanthoxylum Piperitum Fruit Extract)、白头翁根萃取物(Pulsatilla Koreana Root Extract)以及青苔萃取物(Usnea Barbata Extract),业界称为EURO-NApre。于一实施例中,消炎剂或防腐剂的含量为0.1-8%。
于一实施例中,本发明的用以抑制阴道炎病原菌的组合物还包含凝胶剂。举例而言,凝胶剂可为天然聚葡糖胶、卡波姆(Carbomer)或以上的组合。于一实施例中,凝胶剂的含量为0.2-5%。
于一实施例中,本发明的用以抑制阴道炎病原菌的组合物还包含有效成分。举例而言,有效成分可为透明质酸钠(sodium hyaluronate)、抗敏复合物、尿囊素(llantoin)或以上的组合,其中抗敏复合物包含海洋深层水以及西葫芦籽萃取物,业界称为ocaline XP。于一实施例中,有效成分的含量为0.005-5%。
于一实施例中,赋形剂、稀释剂或载体包含氢氧化钠、三乙醇胺、甘油或以上的组合,其含量为0.05-10%,另以纯水作为溶剂补充至100%。
实施例1:本发明的乳酸菌菌株的形态学以及一般性质
根据16S rDNA序列分析以及API细菌鉴定系统分析结果来确认菌株在分类学上的特征。本发明的乳酸菌菌株在形态学及一般性质上的特征详细列于表2:
表2本发明的乳酸菌菌株的形态学及一般性质特征
实施例2:本发明的乳酸菌发酵物的收集
本发明的乳酸菌发酵物是以包含牛奶、奶粉、酪蛋白或以上的组合为基底的培养基,并以经分离的唾液乳酸杆菌AP-32菌株、鼠李糖乳酸杆菌F-1菌株、鼠李糖乳酸杆菌GL-165菌株、动物双歧杆菌乳亚种CP-9菌株、两歧双歧杆菌Bf-688菌株、短双歧杆菌Bv-889菌株或以上乳酸菌菌株的组合与嗜热链球菌共同进行发酵,再经由离心取得发酵液。需说明的是,嗜热链球菌是作为辅助发酵的用途,故未进一步限定特定菌株。依据需求,可进一步将乳酸菌发酵液干燥成乳酸菌发酵物粉末。
实施例3:本发明的乳酸菌的抑制阴道炎病原菌分析
乳酸菌的抑制阴道炎病原菌能力的检测方法是参考2005年Strus等人发表于Candida.Infectious Diseases in Obstetrics and Gynecology,13(2),69-75所使用的抗菌方法进行测试。首先,将活化至第3代的乳酸菌的唾液乳酸杆菌AP-32菌株、罗伊氏乳酸杆菌GL-104菌株、副干酪乳酸杆菌GL-156菌株、鼠李糖乳酸杆菌F-1菌株、鼠李糖乳酸杆菌GL-165菌株、动物双歧杆菌乳亚种CP-9菌株、两歧双歧杆菌Bf-688菌株、短双歧杆菌Bv-889菌株以上乳酸菌菌株个别涂抹在相对应的固态培养基上并维持涂抹宽度为2cm。以嗜热链球菌(Streptococcus thermophilus)SY-66菌株作为负向控制组,控制组(control)则不涂抹乳酸菌。以乳酸菌相对应的培养条件培养2天后,注入14mL用以培养阴道炎病原菌的培养基,待培养基凝固后,将阴道炎病原菌的菌液分别均匀涂抹于培养基上。受测的阴道炎病原菌包含白色念珠球菌(Candida albicans)、金黄色葡萄球菌(Staphylococcus aureus)、大肠杆菌(Escherichia coli)、金黄色葡萄球菌带抗药性菌株(Methicillin-resistantStaphylococcus aureus,MRSA)、B形链球菌(Group B Streptococcus agalactiae,GBS)以及大肠杆菌带抗药性菌株(Escherichia coli-ESBL)。将涂抹阴道炎病原菌的培养基以病原菌相对应的培养条件培养。当到达相应的培养时间后,将菌盘取出,以尺量测抑菌宽度,以评比乳酸菌菌株发酵物的抑菌力。分数越高者抑菌力越好,评价标准如表3所示。
表3评价标准
代表符号 | 抑菌宽度(cm) | 评分 |
(-) | 0 | 0 |
(+/-) | <2 | 1 |
(++) | <3 | 2 |
(+++) | <4 | 3 |
(++++) | <5 | 4 |
(+++++) | <9 | 5 |
(++++++) | 9 | 6 |
本发明的乳酸菌菌株代谢物抑制阴道病原菌的结果如图1以及图2所示。由图1以及图2的结果可知,本发明的乳酸菌菌株代谢物具有抑制阴道炎病原菌的活性效果。
实施例4:本发明的乳酸菌发酵物的抑制阴道炎病原菌分析(一)
乳酸菌发酵物的抑制阴道炎病原菌能力的检测方法如下。于一实施例中,以牛奶、奶粉或酪蛋白为基底的培养基以唾液乳酸杆菌AP-32菌株、鼠李糖乳酸杆菌F-1菌株、GL-165菌株、动物双歧杆菌乳亚种CP-9菌株、两歧双歧杆菌Bf-688菌株、短双歧杆菌Bv-889菌株与嗜热链球菌共同进行发酵,经离心后取出上层发酵液。
以本发明的乳酸菌发酵液作为阴道炎病原菌培养基粉末的溶液,另以未经乳酸菌发酵的上清液调制培养基作为控制组。配制完成后进行灭菌并铸胶半个培养皿。待胶体凝固后,将含控制组的胶体倒入另一半培养皿。接着,将阴道炎病原菌(白色念珠球菌、金黄色葡萄球菌、大肠杆菌)的菌液分别均匀涂抹于上述培养基上。以对应阴道炎病原菌的培养条件进行培养。达到应培养时间后,将菌盘取出,与控制组比较本发明的乳酸菌发酵液的抑菌表现,结果如图3所示。由图3的结果可知,本发明的乳酸菌发酵液的抑制阴道炎病原菌的效果优于控制组。
实施例5:本发明的乳酸菌发酵物的抑制阴道炎病原菌分析(二)
另一乳酸菌发酵物的抑制阴道炎病原菌能力的检测方法如下。将包含10%本发明的乳酸菌发酵液的培养液定量至4.9mL养菌管,另以未经乳酸菌发酵的液态培养基作为控制组。接着,分别加入已活化的阴道炎病原菌(白色念珠球菌、金黄色葡萄球菌、大肠杆菌、金黄色葡萄球菌带抗药性菌株(MRSA)、B形链球菌(GBS)、大肠杆菌带抗药性菌株(ESBL)的菌液0.1mL至养菌管。依照相对应阴道炎病原菌的培养条件进行培养。培养后依适当序列稀释,将共同培养组与控制组序列稀释涂盘于阴道炎病原菌相对应的固态培养基上。将上述菌盘依照相对应阴道炎病原菌的培养条件培养于培养箱中。达到相对应培养时间后,取出菌盘并计数病原菌菌落数以比较抑菌率。抑菌率的计算方法如下:
抑菌率(%)=(1-实验组/控制组)×100%。
本发明乳酸菌发酵物的抑制阴道炎病原菌的检测结果如图4所示,其中符号***表示p值<0.001,亦即统计学上具有非常显著的差异。由图4的结果可知,本发明的乳酸菌发酵液对于阴道炎病原菌皆具有显著的抑制效果,且对于各种阴道炎病原菌的抑菌率皆达80%以上。
实施例6:本发明的乳酸菌发酵物凝胶的抑制阴道炎病原菌分析
乳酸菌发酵物凝胶的抑制阴道炎病原菌能力的检测方法如下。首先,按适当比例称取凝胶剂(例如天然葡糖胶、卡波姆)、有效成分(例如透明质酸钠、Ocaline XP、尿囊素)与50%水搅拌至完全溶解。再加入本发明的乳酸菌发酵液(2%、5%及10%)、消炎剂或防腐剂(例如activonol-M、acnebusters、EURO-Napre、薄荷醇油、茶树精油、芦荟萃取液、左手香萃取物、纳米银)并搅拌混合均匀。之后再加入赋形剂(例如氢氧化钠、三乙醇胺、甘油)并调整pH值至4.5-6.5,并以纯水补至100%,再搅拌均匀。控制组仅有凝胶剂而不含本发明的乳酸菌发酵液、有效成分以及消炎剂/防腐剂;另一对照组则仅含有凝胶剂以及消炎剂/防腐剂但不含本发明的乳酸菌发酵液。各别取0.1ml已活化的阴道炎病原菌(浓度为1×105~9×105CFU/ml的白色念珠球菌、金黄色葡萄球菌、大肠杆菌)加入2.5g各组实验凝胶中混合,混和均匀后静置20分钟。接着,分别添加22.5ml的纯水稀释10倍后,再依适当序列稀释后取0.1ml进行涂盘并于37℃培养。最后依据前述的抑菌率计算方法求得抑菌率。
本发明的乳酸菌发酵物凝胶的抑制阴道炎病原菌分析结果如图5所示,其中符号***表示p值<0.001,符号**表示p值<0.01,亦即统计学上具有非常显著的差异;符号*表示p值<0.05,亦即统计学上具有显著差异。由图5的结果可知,仅含有消炎剂/防腐剂的对照组相较于控制组虽具统计意义,但其对于阴道病原菌的抑菌率皆低于75%。而本发明的含有2%,5%及10%乳酸菌发酵液的凝胶对于阴道炎病原菌皆具有显著的抑制效果,抑菌率达90%以上,其中本发明的含有5%及10%乳酸菌发酵液的凝胶对于各种阴道炎病原菌的抑菌率皆达100%。
综合上述,本发明的用以抑制阴道炎病原菌的组合物包含乳酸菌发酵物,其是于包含牛奶、奶粉、酪蛋白至少其中之一的培养基中以唾液乳酸杆菌AP-32菌株、鼠李糖乳酸杆菌F-1菌株、鼠李糖乳酸杆菌GL-165菌株、动物双歧杆菌乳亚种CP-9菌株、两歧双歧杆菌Bf-688菌株、短双歧杆菌Bv-889菌株或以上乳酸菌菌株的组合与嗜热链球菌共同进行发酵所得。上述乳酸菌菌株发酵物可抑制阴道炎病原菌生长,因此可作为抑制阴道炎病原菌的用途,并以食品组合物、医药组合物或阴道清洁组合物的形式存在。较佳者,本发明的用以抑制阴道炎病原菌的组合物还包含消炎剂/防腐剂或其它有效成分,以进一步提升抑制阴道炎病原菌的能力。
以上所述的实施例仅是为说明本发明的技术思想及特点,其目的在使熟习此项技艺的人士能够了解本发明的内容并据以实施,当不能以之限定本发明的专利范围,即大凡依本发明所揭示的精神所作的均等变化或修饰,仍应涵盖在本发明的专利范围内。
Claims (7)
1.一种用以抑制阴道炎病原菌的外用剂型医药组合物,其包含:
乳酸菌发酵物,其是以包含牛奶、奶粉及酪蛋白至少其中之一的培养基,并以经分离的乳酸菌菌株与嗜热链球菌共同进行发酵而获得,其中所述乳酸菌菌株包含保藏编号为CCTCC NO:M2011127的唾液乳酸杆菌(Lactobacillus salivarius subsp. salicinius)AP-32菌株、保藏编号为CCTCC NO:M2011124的鼠李糖乳酸杆菌(Lactobacillus rhamnosus) F-1菌株、保藏编号为CCTCC NO:M2014591的鼠李糖乳酸杆菌(Lactobacillus rhamnosus) GL-165菌株、保藏编号为CCTCC NO:M2014588的动物双歧杆菌乳亚种(Bifidobacterium animalis subsp. lactis) CP-9菌株、保藏编号为CGMCC No. 17953的两歧双歧杆菌(Bifidobacterium bifidum) Bf-688菌株、以及保藏编号为CGMCC No.16145的短双歧杆菌(Bifidobacterium breve) Bv-889菌株,上述乳酸菌菌株分别保藏于中国典型培养物保藏中心以及中国微生物菌种保藏管理委员会普通微生物中心,其中所述乳酸菌发酵物的含量为2%-10%;以及
赋形剂。
2.如权利要求1所述的用以抑制阴道炎病原菌的外用剂型医药组合物,其还包含消炎剂或防腐剂。
3.如权利要求1所述的用以抑制阴道炎病原菌的外用剂型医药组合物,其还包含凝胶剂。
4.如权利要求3所述的用以抑制阴道炎病原菌的外用剂型医药组合物,其中所述凝胶剂包含天然聚葡糖胶、卡波姆(Carbomer)或以上的组合,且凝胶剂的含量为0.2%-5%。
5.如权利要求1所述的用以抑制阴道炎病原菌的外用剂型医药组合物,其还包含有效成分,有效成分包含透明质酸钠(sodium hyaluronate)、抗敏复合物、尿囊素(allantoin)或以上的组合,且有效成分的含量为0.005%-5%,其中抗敏复合物包含海洋深层水以及西葫芦籽萃取物。
6.如权利要求1所述的用以抑制阴道炎病原菌的外用剂型医药组合物,其中所述赋形剂包含氢氧化钠、三乙醇胺、甘油或以上的组合,其含量为0.05%-10%,其余以纯水补充至100%。
7.一种阴道清洁组合物,其包含:
乳酸菌发酵物,其是以包含牛奶、奶粉以及酪蛋白至少其中之一的培养基,并以经分离的乳酸菌菌株与嗜热链球菌共同进行发酵而获得,其中所述乳酸菌菌株包含保藏编号为CCTCC NO:M2011127的唾液乳酸杆菌(Lactobacillus salivarius subsp. salicinius)AP-32菌株、保藏编号为CCTCC NO:M2011124的鼠李糖乳酸杆菌(Lactobacillus rhamnosus) F-1菌株、保藏编号为CCTCC NO:M2014591的鼠李糖乳酸杆菌(Lactobacillus rhamnosus) GL-165菌株、保藏编号为CCTCC NO:M2014588的动物双歧杆菌乳亚种(Bifidobacterium animalis subsp. lactis) CP-9菌株、保藏编号为CGMCC No. 17953的两歧双歧杆菌(Bifidobacterium bifidum) Bf-688菌株、以及保藏编号为CGMCC No.16145的短双歧杆菌(Bifidobacterium breve) BV-889菌株,上述乳酸菌菌株分别保藏于中国典型培养物保藏中心以及中国微生物菌种保藏管理委员会普通微生物中心,其中所述乳酸菌发酵物的含量为2%-10%;以及
生理上可接受的赋形剂。
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CN114480191A (zh) * | 2022-01-21 | 2022-05-13 | 河北源民生物科技有限公司 | 一种乳杆菌复合菌粉在防治阴道炎制品中的应用方法 |
CN115161348B (zh) * | 2022-04-28 | 2024-09-13 | 海普诺凯营养品有限公司 | 一种具有多功效的后生元组合物 |
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