CN115137758A - 一种有利于女性生殖道健康的益生菌的制备及其应用 - Google Patents
一种有利于女性生殖道健康的益生菌的制备及其应用 Download PDFInfo
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Abstract
本发明涉及一种有利于女性生殖道健康的益生菌的制备及其应用。具体地,本发明提供,含有保藏编号CGMCC NO.16871的植物乳杆菌,在制备有利于女性生殖道健康的药品、食品或保健品中的应用。所述植物乳杆菌用于清除自由基,抗氧化,抑制病原菌生长,调节肠道菌群,刺激免疫系统,减少炎症发生,从而保护女性私密健康。本发明提供的益生菌制剂激发宿主免疫防御的级联反应,促进上皮细胞产生粘液,降低促炎因子IL‑4、IL‑1β和TNF‑α的含量,减轻炎症损伤,从而发挥抵抗疾病的作用,维持阴道的健康环境。
Description
技术领域
本发明涉及益生菌技术领域,具体涉及一种有利于女性生殖道健康的的益生菌的制备及其应用。
背景技术
一般来说,健康的女性阴道内存在着200多种微生物,阴道内的微生物以乳杆菌为主,占微生物总量的90-95%,阴道的酸性环境就是由这些乳杆菌来维持的,酸性环境可以抑制病原菌的生长。除此之外,还有大肠埃希菌、衣原体及念珠菌等,这些微生物共同维持着阴道的微生态平衡。
现代女性由于生活压力的不断增加,饮食作息的不合理化,卫生情况以及激素水平的变化导致体内微生态系统紊乱,乳杆菌减少,致病菌如大肠埃希菌、葡萄球菌等数量增加,打破了阴道的菌群平衡,引起感染,导致会引起多种疾病如细菌性阴道炎、外阴阴道假丝酵母菌病、需氧菌性阴道病等。细菌性阴道炎感染严重者可引起多种疾病比如慢性宫颈炎、宫颈癌、卵巢癌、输卵管肿瘤等疾病;同时也是造成不孕、早产、复发性流产等的主要原因。据调查,中国患有妇科疾病的人群占到了女性总人口的87.3%,妇科疾病严重影响了女性正常的生活和工作。
对于女性阴道健康问题,目前传统的治疗方法是通过抗生素治疗,虽然效果显著,但是基本都有副作用,抗生素虽然能减轻炎症,但会破坏正常的阴道菌群结构,因而容易复发;同时,由于抗菌药物的不合理使用,容易导致病原菌产生抗药性。因此,微生物疗法对于女性阴道疾病的治疗具有重大意义。
益生菌是一类有益于宿主的活的微生物,能促进人体的菌群生态平衡。双歧杆菌与乳酸菌在宿主消化、免疫调节以及慢性疾病的预防与治疗方面发挥着重要作用。与抗生素相比,益生菌对于宿主不会有副作用,而且不会产生抗药性。益生菌在宿主机体内生长繁殖过程中分泌的代谢产物有机酸,能降低阴道内的pH值,阴道在酸性环境下,会抑制或杀灭其他细菌的生长,最终达到自净的能力。同时,益生菌能产生过氧化氢,促进具有过氧化物酶活性的蛋白抑制剂与H2O2结合或直接通过H2O2抑制致病菌的生长,并且与致病菌竞争养分和竞争对阴道黏膜的占位。与此同时,过氧化氢还能清除阴道内的自由基,减轻氧化反应。当阴道内的菌群失衡时,会产生一系列的炎症反应,导致细菌性阴道炎等疾病的发生,益生菌可以激发宿主免疫防御的级联反应,促进上皮细胞产生粘液,降低促炎因子IL-6、IL-1β和TNF-α的含量,减轻炎症损伤,从而发挥抵抗疾病的作用,维持阴道的健康环境。
发明内容
本发明的目的是提供一种用于改善女性阴道健康的益生菌制剂,具体地,本发明的目的是提供一株具有抑制女性生殖道致病菌的植物乳杆菌及其应用。
本发明请求保护益生菌在制备抑制女性生殖道致病菌的药品、食品或保健品中的应用;所述益生菌为保藏编号CGMCC NO.16871的植物乳杆菌。
在本发明提供的应用中,所述植物乳杆菌用于清除自由基、抗氧化,抑制病原菌生长,调节肠道菌群,刺激免疫系统,减少炎症发生。
在本发明提供的应用中,所述益生菌为保藏编号CGMCC NO.16871的植物乳杆菌发酵后的活菌或其发酵产物。
在本发明提供的应用中,所述益生菌制剂为植物乳杆菌冻干粉。
在本发明提供的应用中,保藏编号CGMCC NO.16871的植物乳杆菌的发酵方法,包括:
(1)将保藏的甘油管菌种以3%(v/v)接种量接种至种子培养基中,在37℃条件下静置厌氧条件培养得到一级种子液;
(2)将(1)制备的一级种子液以3%(v/v)接种量接种至种子培养基中,在37℃条件下静置厌氧条件培养得到二级种子液;
(3)将(2)培养的二级种子液按照5%(V/V)的接种量接至发酵培养基中,于37℃条件下静置厌氧条件培养,得到发酵液;
将发酵液进行离心收集上清液,即为植物乳杆菌发酵产物。
植物乳杆菌的发酵方法步骤(1)和(2)中,所述种子培养基独立包括以下质量百分含量的组分:1~3%葡萄糖、0.5~2%蛋白胨、0.5~2%酵母浸粉、0.2~1%牛肉浸粉,0.2~1%无水乙酸钠、0.1~0.5%柠檬酸铵、0.1~0.5%磷酸氢二钾,0.05~0.2%吐温80和余量的水,pH值为6.7~6.9。
植物乳杆菌活菌的制备方法步骤(3)所述发酵培养基,按质量分数计,包括1~3%葡萄糖、0.5~2%蛋白胨、0.5~2%酵母浸粉、0.2~1%牛肉浸粉,0.2~1%无水乙酸钠、0.1~0.5%柠檬酸铵、0.1~0.5%磷酸氢二钾,0.05~0.2%吐温80和余量的水,pH值为6.3~6.5。
植物乳杆菌活菌的制备方法步骤(3)中,发酵温度为36~38℃,所述高密度发酵时间7~11h。
保藏编号CGMCC NO.16871的植物乳杆菌冻干粉的制备方法,包括:
(1)将所述植物乳杆菌的发酵液离心取沉淀,得到菌泥;
(2)将菌泥与冻干保护剂混合后进行乳化包埋,得到乳化液;
(3)将得到的乳化液冷冻干燥,粉碎,得到植物乳杆菌冻干粉。
本发明提供的益生菌为植物乳杆菌冻干粉,所述菌泥与冻干保护剂的质量比为1:0.5~2;所述乳化包埋时间为30~60min。
在本发明提供的应用中,其中冻干粉的活菌数为2000~3000亿CFU/g。
根据本领域技术人员的理解,本发明请求保护含有保藏编号CGMCC NO.16871的植物乳杆菌的益生菌菌粉在制备治疗大鼠阴道炎的药物中的应用。以及含有保藏编号CGMCCNO.16871的植物乳杆菌的复合益生菌制剂的制备方法和在制备防治女性细菌性阴道炎的药物中的应用。
本发明所采取的技术方案是:
一种含有植物乳杆菌CGMCC NO.16871的复合益生菌制剂,其组成包括:植物乳杆菌CGMCC NO.16871菌粉3.4%、鼠李糖乳杆菌nbk-W120 1%、罗伊氏乳杆菌nbk-LRe90.3%、乳双歧杆菌nbk-W130.5%、嗜酸乳杆菌nbk-W563 1%、乳糖醇32%、抗性糊精25%、菊粉17%、低聚木糖12%、蔓越莓果粉6%、清蛋白肽1%、玫瑰茄提取物0.5%、蔓越莓提取物0.3%。
所述含有植物乳杆菌CGMCC NO.16871的复合益生菌制剂对原料进行了优选。其中添加的植物乳杆菌CGMCC NO.16871分离自中国传统泡菜,具有抗氧化功能及降低炎症因子分泌,提高免疫力的功效鼠李糖乳杆菌nbk-W120来源于健康婴儿粪便,适合国人体质,不仅可以调节肠道菌群,定殖能力强,还具有抗氧化的功能。罗伊氏乳杆菌nbk-LRe9是从健康女性的肠道中筛选的功能性菌株,它可以改善肠道菌群,具有很强的抑制有害菌定植的能力,能广泛抑制革兰氏阳性菌、革兰氏阴性菌、酵母、真菌和病原虫等的生长,呵护女性私密健康功能。嗜酸乳杆菌nbk-W563筛选自健康婴儿肠道。可产生多种有机酸,维持阴道酸性环境,同时具有抗氧化能力,有效防止过氧化损伤。乳双歧杆菌nbk-W13筛选自健康婴儿肠道。可以增强粘膜免疫屏障功能,修复粘膜损伤,调节抗炎因子及炎症因子的平衡,减少炎症反应的发生。
通过五种复合菌粉的复配能达到协同增效的作用。低聚木糖和抗性糊精为可溶性膳食纤维,可以促进阴道微生态平衡,抑制有害菌生长,减少毒素产生,增强机体免疫力。清蛋白肽为小分子肽,易于吸收,能促进植物乳杆菌CGMCC NO.16871的增殖。通过复合菌粉中各组分以特定比例进行复配,能使功效达到最大。
本发明的有益效果在于,采用本发明的制备方法得到的植物乳杆菌CGMCCNO.16871具有优良的耐酸耐胆盐能力,不仅不会有副作用,而且还能产生有机酸,降低阴道内的pH值,抑制病原菌的生长,促进阴道的菌群平衡。同时还能刺激免疫系统,增强阴道的免疫功能,产生过氧化氢,清除自由基,从而减少炎症反应,维持阴道的健康环境。采用本发明的制备方法得到的含有植物乳杆菌CGMCC NO.16871的复合益生菌制剂,具有缓解女性阴道瘙痒,改善白带异常,调节阴道环境pH值,减少异味等作用,呵护女性私密健康。
附图说明
图1是本发明植物乳杆菌CGMCC NO.16871抑制致病菌生长平板图;其中,A为金黄色葡萄球菌的抑菌圈;B为大肠埃希氏菌的抑菌圈;C为白假丝酵母的抑菌圈。
图2是本发明植物乳杆菌CGMCC NO.16871对DPPH自由基的清除率。
图3是本发明植物乳杆菌CGMCC NO.16871对羟自由基的清除率。
图4是本发明植物乳杆菌CGMCC NO.16871对超氧离子的清除率。
图5是本发明植物乳杆菌CGMCC NO.16871对亚铁离子的螯合能力。
图6是本发明植物乳杆菌CGMCC NO.16871总还原能力。
图7是植物乳杆菌CGMCC NO.16871对大鼠阴道内菌群的影响。
图8为清蛋白肽促进植物乳杆菌CGMCC NO.16871增殖的作用。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。
实施例1植物乳杆菌CGMCC NO.16871的筛选及鉴定
植物乳杆菌分离自中国传统泡菜,经研磨,无菌生理盐水稀释至合适梯度,于MRS琼脂平板上采用四区划线法划线后,37℃培养箱中厌氧培养48h,挑选单菌落,镜检,划线,进一步纯化经鉴定为植物乳杆菌。植物乳杆菌的生理生化特性见表1。
表1细胞形态及生理生化结果
16S rRNA基因序列(SEQ ID NO.1)测定结果如下:
实施例2植物乳杆菌CGMCC NO.16871的发酵方法
以质量百分比计,本实施例所用种子培养基为2%葡萄糖、2%蛋白胨、0.5%酵母浸粉、1%牛肉浸粉,0.5%无水乙酸钠、0.2%柠檬酸铵、0.2%磷酸氢二钾,0.1%吐温80和余量的水,pH值为6.8。
以质量百分比计,本实施例所用发酵培养基为2%葡萄糖、2%蛋白胨、0.5%酵母浸粉、1%牛肉浸粉,0.5%无水乙酸钠、0.2%柠檬酸铵、0.2%磷酸氢二钾,0.1%吐温80和余量的水,pH值为6.4。
本实施例提供,植物乳杆菌CGMCC NO.16871的发酵方法,步骤如下:
(1)将保藏的甘油管菌种以3%(v/v)接种量接种至种子培养基中,在37℃条件下静置厌氧条件培养18h得到一级种子液。
(2)将(1)制备的一级种子液以3%(v/v)接种量接种至种子培养基中,在36℃条件下静置厌氧条件培养15h得到二级种子液。
(3)将(2)培养的二级种子液按照5%(V/V)的接种量接至发酵培养基中,于37℃条件下静置厌氧条件培养8h,得到发酵液。
将发酵液进行离心收集上清液,即为植物乳杆菌发酵产物,离心条件为4000-8000rpm离心20-30min。
实施例3植物乳杆菌CGMCC NO.16871活菌体的制备
植物乳杆菌CGMCC NO.16871菌粉的制备方法,包括以下步骤:
(1)将实施例2得到的发酵液离心取沉淀,得到菌泥;
(2)将菌泥与冻干保护剂混合后进行乳化包埋,得到乳化液;
(3)将得到的乳化液冷冻干燥,粉碎,得到植物乳杆菌冻干粉。
按质量百分比计,冻干保护剂:脱脂乳10%,甘油6%,其余量为水。
待发酵液中菌体密度达到109cfu/mL,收集发酵液,离心获得菌体,将菌体加入保护剂进行乳化包埋,乳化时间30min;将乳化液在-38℃下预冷冻2h,将预冷冻后的乳化液进行真空冷冻干燥,真空度为0.2mba;真空冷冻干燥后将其粉碎至20目,得到植物乳杆菌制剂,所得植物乳杆菌CGMCC NO.16871制剂的活菌数为2.0×1011cfu/g。
实施例4植物乳杆菌CGMCC NO.16871模拟人工胃肠液和耐胆盐实验
将划线纯化后保存的植物乳杆菌CGMCC NO.16871传代以后,以1%的接种量接种于pH2.5的人工胃液中,处理2小时后再以1%的接种量接种于pH8.0的人工肠液中处理6小时,采用平板计数法进行活菌计数,结果见表2。
表2植物乳杆菌CGMCC NO.16871在模拟人工胃肠液中的存活率
由表2可看出,植物乳杆菌CGMCC NO.16871在pH2.5的人工胃液中处理2小时后存活率可达到91.86%,经pH8.0的人工肠液处理6小时后活菌数有所增长。说明植物乳杆菌在进入人体后可顺利通过胃肠道。
将划线纯化后保存的植物乳杆菌CGMCC NO.16871传代以后,以1%的接种量分别接种于胆盐浓度为0.3%、0.6%、0.9%、1.2%的MRS培养基中,处理6小时后,采用平板计数法进行活菌计数,结果见表3。
表3植物乳杆菌CGMCC NO.16871在不同浓度胆盐中的存活率
由表3可看出,植物乳杆菌CGMCC NO.16871在1.2%的胆盐浓度下仍能存活,说明植物乳杆菌CGMCC NO.16871具有良好的胆盐耐受性。
实施例5植物乳杆菌CGMCC NO.16871体外抑菌实验
将划线纯化后保存的菌株植物乳杆菌CGMCC NO.16871接种于装有5mLMRS液体培养基的试管中,37℃静置培养24h。发酵液经8000r/min离心3min,取上清液。
采用牛津平板法测定菌株是否具有抑菌活性:将活化后的致病菌与加热融化的固体培养基混匀,使菌落终浓度至107CFU/mL,倾注约15mL于无菌平皿中,待培养基凝固后,放入牛津杯,取200μL上清液加入牛津杯,4℃条件下扩散4h后,37℃条件下培养10h,观察抑菌圈。采用十字交叉法测量抑菌圈直径,以未接种的培养基上清液作为空白对照,重复3次。当抑菌圈直径≥4.0mm视为有抑菌作用。结果见图1,表4。
表4植物乳杆菌CGMCC NO.16871抑制致病菌抑菌圈大小
+++:抑菌圈直径≥20mm,表示极抑菌;++:15mm≤抑菌圈直径<20mm,表示高度抑菌;+:7mm≤抑菌圈直径<15mm,表示一般抑菌;-:抑菌圈直径<7mm,表示不抑菌。
由图1和表4可看出,植物乳杆菌CGMCC NO.16871的菌液对金黄色葡萄球菌表现出极抑菌;菌液和上清液均对大肠埃希氏菌表现出高度抑菌;菌液和菌体对白假丝酵母表现出抑菌效果,综合结果说明,植物乳杆菌CGMCC NO.16871对金黄色葡萄球菌、大肠埃希氏菌和白假丝酵母表现出抑菌效果,不仅发酵产物具有抑菌效果,其菌体也有明显的抑菌效果,证明其死菌也有抑菌效果。
实施例6植物乳杆菌CGMCC NO.16871体外抗氧化实验
将划线纯化后保存的菌株植物乳杆菌CGMCC NO.16871接种于装有5mLMRS液体培养基的试管中,37℃静置培养24h。发酵液经8000r/min离心3min,取上清液。
(1)DPPH自由基清除能力的测定:取样品1mL置于试管中,加入DPPH无水乙醇溶液2m L(浓度为0.2mmol/L),混匀后,室温下避光反应30min,4℃,8000g,离心10min后取上清液,517nm下测定其吸光度,去离子水调零。每个样品3个重复。
(2)OH自由基清除能力的测定:1ml O-菲啰啉(浓度为0.1%)中加入样品0.5ml,PBS 1mL,2.5mmol/L FeSO4 1mL,20mmol/L H2O21mL。37℃恒温水浴1.5h后,536nm下测吸光度。每个样品3个重复。
(3)O2 -清除能力的测定:取2.8mL Tris-HCl(0.05mol/L,pH 8.2)置于试管中,加入0.1mL邻苯三酚(0.05mol/L),0.1mL样品。混匀后25℃,黑暗中反应4min。然后添加1mL8mol/L HCl终止反应,用多功能酶标仪在320nm下检测吸光度。每个样品3个重复。
(4)Fe2+螯合能力的测定:1%抗坏血酸0.1mL,0.4%FeSO4 0.1mL,NaOH(0.2mol/L)1mL的混合液加入0.5mL的乳酸菌的待测液,混合液37℃水浴20min,三氯乙酸沉淀蛋白,3000×g、4℃离心10mim,取0.2mL上清液加入2mL 0.1%邻-菲罗啉反应10min,于510nm测吸光值,以磷酸盐缓冲液(PBS 0.2mol/L,pH6.6)作为空白对照。
(5)总还原能力的测定:取0.5m L样品置于试管中,加入浓度为0.2mol/L pH 6.6的磷酸缓冲溶液0.5m L,再加入0.5m L 1%铁氰化钾,50℃水浴20min后迅速冰浴冷却。再加入10%三氯乙酸0.5m L,4000r/min离心10min,取上清液1m L,加入1mL蒸馏水和1mL0.1%三氯化铁,混合均匀。室温放置10min,700nm波长测定其吸光度。
由图2、图3、图4、图5、图6可看出,植物乳杆菌CGMCC NO.16871对于DPPH和羟基自由基的清除率效果明显,上清液和菌体对DPPH的清除率均达到65%以上;对羟自由基的清除率可达到30%以上;上清液对超氧离子的清除率大于45%,菌体的清除率效果不明显;上清液和菌体对Fe2+螯合率均达到了20%以上;上清液的总还原能力为56%,菌体为33%。说明植物乳杆菌CGMCC NO.16871不仅发酵产物具有抗氧化、清除自由基的效果,其菌体也有明显的效果,为小鼠细菌性阴道炎的治疗提供了前提。
实施例7植物乳杆菌CGMCC NO.16871改善大鼠细菌性阴道炎的研究
1、大鼠阴道炎模型的建立:
(1)实验分组为空白对照组、模型组、实验组,每组10只大鼠。
(2)将白色念珠菌、金黄色葡萄球菌以及大肠杆菌按照1:1:2的比例配制成3×1010的混悬液,对实验大鼠造模,空白对照组为PBS处理。造模期间解剖部分大鼠取子宫组织做HE染色,判断造模是否成功。
2、植物乳杆菌对大鼠阴道炎模型的治疗:
(1)造模成功后进行7天给药治疗:将实施例3制得的植物乳杆菌CGMCC NO.16871冻干粉用无菌PBS配制成1×1011CFU/mL的菌液,用枪头吸取50μL菌液,放入大鼠阴道内,连续给药7天,对照组为PBS处理。恢复一周进行后续实验;(2)恢复期后,颈椎脱臼处死大鼠,解剖,取大鼠子宫组织;采用Elisa法检测大鼠子宫组织中IL-4、IL-1β、IFN-γ及TNF-α炎性因子蛋白水平,见表5。
(2)测定大鼠阴道菌群种类及数量的变化。见图7。
表5大鼠阴道中TNF-α、IL-4、IL-1β、IFN-γ水平的变化
表中*代表相比于模型组P<0.05,**代表相比于模型组P<0.01
细胞因子是机体活化的淋巴细胞产生的免疫活性物质,具有免疫调节特性,在免疫系统中发挥重要的作用。细胞因子分泌量的多少,在一定程度上能够反映机体免疫系统免疫应答的水平和状态。表5的实验结果表明,植物乳杆菌CGMCC NO.16871菌粉能显著增加小鼠阴道中抑炎因子IL-4的含量,能显著降低促炎因子TNF-α、IFN-γ和IL-1β的含量,表明植物乳杆菌CGMCC NO.16871菌粉对大鼠阴道细胞因子的含量有一定的调节作用,能减轻炎症反应,抑制细菌性阴道炎,维持阴道的健康。
大鼠阴道菌群中有益菌和有害菌的数量影响着阴道菌群的平衡与阴道健康。由图7可看出,与模型组大鼠相比,实验组大鼠阴道菌群中乳杆菌、双歧杆菌数量显著增加,肠杆菌、肠球菌数量显著减少,基本恢复到正常组水平。说明,植物乳杆菌CGMCC NO.16871能改善细菌性阴道炎大鼠的阴道菌群,促进有益菌乳杆菌和双歧杆菌的增殖,抑制有害菌肠杆菌、肠球菌的生长,维持阴道健康。
实施例8含有植物乳杆菌CGMCC NO.16871的固体饮料的制备
本实施例的复合益生菌制剂的配方如下:植物乳杆菌CGMCC NO.16871菌粉3.4%、鼠李糖乳杆菌nbk-W120 1%、罗伊氏乳杆菌nbk-LRe9 0.3%、乳双歧杆菌nbk-W13 1%、嗜酸乳杆菌nbk-W563 0.5、乳糖醇32%、抗性糊精25%、菊粉17%、低聚木糖12%、蔓越莓果粉6%、清蛋白肽1%、玫瑰茄提取物0.5%、蔓越莓提取物0.3%。
制备方法如下:
生产过程中环境温度控制不高于25℃,湿度控制在40%左右。
烘干:低聚木糖、抗性糊精、菊粉需先测定水分,其中水分含量>4%的物料要进行烘箱干燥法烘料预处理,参数为60℃,3h,烘干后检测水分<5%;
过筛:乳糖醇过60目过筛备用。
菌粉回温及过筛:混料前-18℃储存的菌粉取出放置室温1h,使菌粉回温至常温状态。
混料:将蔓越莓果粉、提取物及清蛋白肽与植物乳杆菌CGMCC NO.16871、鼠李糖乳杆菌nbk-W120、罗伊氏乳杆菌nbk-LRe9、嗜酸乳杆菌nbk-W563、乳双歧杆菌nbk-W13混合均匀,得混合均匀的小料,小料混料时间不超过20min,再将菊粉、抗性糊精、低聚木糖、乳糖醇按顺序依次加入,混料时间不超过20min。
装袋:混合粉放入洁净无菌条袋机内,设置条袋规格2g/袋进行生产;
清蛋白肽促进植物乳杆菌CGMCC NO.16871生长的情况见图8。
实施例9含有植物乳杆菌CGMCC NO.16871的复合益生菌制剂在防治女性细菌性阴道炎中的应用
将实施例8制得的含有植物乳杆菌CGMCC NO.16871的复合益生菌制剂进行人群试服效果实验。
(1)试验对象:女性细菌性阴道炎患者50名,具体症状为阴道瘙痒、白带分泌异常,阴道分泌物pH值升高,有异味等。随机分为两组。
(2)试验过程:两组参试人员分别口服实施例8制得的含有植物乳杆菌CGMCCNO.16871的复合益生菌制剂(2g/d)及相同剂量不含益生菌的安慰剂,4周后,对每名参试者进行阴道瘙痒、白带异常情况、异味、阴道pH值等情况调查,调查统计结果参见表6、表7、表8、表9。
表6植物乳杆菌CGMCC NO.16871对女性阴道pH值的影响
平均阴道pH值 | 使用前 | 使用后 |
对照组 | 5.22 | 5.06 |
实验组 | 5.34 | 4.68 |
表7植物乳杆菌CGMCC NO.16871对女性阴道瘙痒情况的影响
瘙痒改善情况 | 痊愈/例 | 改善/例 | 无效/例 | 有效率/% | 合计/人 |
对照组 | 1 | 11 | 10 | 54.5 | 22 |
实验组 | 4 | 15 | 2 | 90.5 | 21 |
表8植物乳杆菌CGMCC NO.16871对女性阴道白带异常情况的影响
白带异常改善情况 | 痊愈/例 | 改善/例 | 无效/例 | 有效率/% | 合计/人 |
对照组 | 0 | 8 | 9 | 47.1 | 17 |
实验组 | 6 | 11 | 3 | 85.0 | 20 |
表9植物乳杆菌CGMCC NO.16871对女性阴道分泌物异味的影响
由表6、表7、表8、表9可知,与对照组相比,服用添加了植物乳杆菌CGMCC NO.16871的复合益生菌制剂4周后,患者阴道的pH值明显降低,有利于乳酸菌在阴道内的定殖,同时,阴道瘙痒情况,分泌物异味情况、白带异常情况均有所改善。因此,植物乳杆菌CGMCCNO.16871与鼠李糖乳杆菌、罗伊氏乳杆菌复配后具有协同增效的作用,同时,益生菌与抗性糊精等益生元以及胶原三肽复配也具有协同的作用,可以促进阴道微生态的平衡,改善阴道环境,在防治细菌性阴道炎方面具有良好的效果。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
序列表
<110> 诺佰克(武汉)生物科技有限公司
<120> 一种有利于女性生殖道健康的益生菌的制备及其应用
<130> KHP211124680.5
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1380
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
tctcatggtg tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg cggcatgctg 60
atccgcgatt actagcgatt ccgacttcat gtaggcgagt tgcagcctac aatccgaact 120
gagaatggct ttaagagatt agcttactct cgcgagttcg caactcgttg taccatccat 180
tgtagcacgt gtgtagccca ggtcataagg ggcatgatga tttgacgtca tccccacctt 240
cctccggttt gtcaccggca gtctcaccag agtgcccaac ttaatgctgg caactgataa 300
taagggttgc gctcgttgcg ggacttaacc caacatctca cgacacgagc tgacgacaac 360
catgcaccac ctgtatccat gtccccgaag ggaacgtcta atctcttaga tttgcatagt 420
atgtcaagac ctggtaaggt tcttcgcgta gcttcgaatt aaaccacatg ctccaccgct 480
tgtgcgggcc cccgtcaatt cctttgagtt tcagccttgc ggccgtactc cccaggcgga 540
atgcttaatg cgttagctgc agcactgaag ggcggaaacc ctccaacact tagcattcat 600
cgtttacggt atggactacc agggtatcta atcctgtttg ctacccatac tttcgagcct 660
cagcgtcagt tacagaccag acagccgcct tcgccactgg tgttcttcca tatatctacg 720
catttcaccg ctacacatgg agttccactg tcctcttctg cactcaagtt tcccagtttc 780
cgatgcactt cttcggttga gccgaaggct ttcacatcag acttaaaaaa ccgcctgcgc 840
tcgctttacg cccaataaat ccggacaacg cttgccacct acgtattacc gcggctgctg 900
gcacgtagtt agccgtggct ttctggttaa ataccgtcaa tacctgaaca gttactctca 960
gatatgttct tctttaacaa cagagtttta cgagccgaaa cccttcttca ctcacgcggc 1020
gttgctccat cagactttcg tccattgtgg aagattccct actgctgcct cccgtaggag 1080
tttgggccgt gtctcagtcc caatgtggcc gattaccctc tcaggtcggc tacgtatcat 1140
tgccatggtg agccgttacc tcaccatcta gctaatacgc cgcgggacca tccaaaagtg 1200
atagccgaag ccatctttca aactcggacc atgcggtcca agttgttatg cggtattagc 1260
atctgtttcc aggtgttatc ccccgcttct gggcaggttt cccacgtgtt actcaccagt 1320
tcgccactca ctcaaatgta aatcatgatg caagcaccaa tcaataccag agttcgttcg 1380
Claims (10)
1.益生菌在制备有利于女性生殖道健康的药品、食品或保健品中的应用;所述益生菌为保藏编号CGMCC NO.16871的植物乳杆菌。
2.根据权利要求1所述的应用,其特征在于,所述植物乳杆菌用于清除自由基、抗氧化,抑制病原菌生长,调节肠道菌群,刺激免疫系统,减少炎症发生。
3.根据权利要求2所述的应用,其特征在于,所述益生菌为保藏编号CGMCC NO.16871的植物乳杆菌发酵后的活菌或其发酵产物。
4.根据权利要求3所述的应用,其特征在于,保藏编号CGMCC NO.16871的植物乳杆菌的发酵方法,包括:
(1)将保藏的甘油管菌种以3%(v/v)接种量接种至种子培养基中,在37℃条件下静置厌氧条件培养得到一级种子液;
(2)将(1)制备的一级种子液以3%(v/v)接种量接种至种子培养基中,在37℃条件下静置厌氧条件培养得到二级种子液;
(3)将(2)培养的二级种子液按照5%(V/V)的接种量接至发酵培养基中,于37℃条件下静置厌氧条件培养,得到发酵液;
将发酵液进行离心收集上清液,即为植物乳杆菌发酵产物。
5.根据权利要求4所述的应用,其特征在于,所述益生菌为植物乳杆菌冻干粉。
6.根据权利要求5所述的应用,其特征在于,保藏编号CGMCC NO.16871的植物乳杆菌冻干粉的制备方法,包括:
(1)将所述植物乳杆菌的发酵液离心取沉淀,得到菌泥;
(2)将菌泥与冻干保护剂混合后进行乳化包埋,得到乳化液;
(3)将得到的乳化液冷冻干燥,粉碎,得到植物乳杆菌冻干粉。
7.根据权利要求5-6任一项所述的应用,其特征在于,植物乳杆菌冻干粉的活菌数为2000~3000亿CFU/g。
8.根据权利要求1-2任一项所述的应用,其特征在于,含有保藏编号CGMCC NO.16871的植物乳杆菌的益生菌菌粉在制备治疗大鼠阴道炎的药物或在制备固体饮料中的应用。
10.含有保藏编号CGMCC NO.16871的植物乳杆菌的复合益生菌制剂在制备防治女性细菌性阴道炎的药物中的应用。
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