CN111575211A - 一种用于缓解过敏性鼻炎的副干酪乳杆菌及其应用 - Google Patents
一种用于缓解过敏性鼻炎的副干酪乳杆菌及其应用 Download PDFInfo
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Abstract
本申请公开了一种用于缓解过敏性鼻炎的副干酪乳杆菌及其应用,所述副干酪乳杆菌为副干酪乳杆菌Probio‑37(Lactobacillus paracasei Probio‑37),所述副干酪乳杆菌Probio‑37该菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCC No.18638。所述副干酪乳杆菌能刺激机体Th1/Th2免疫平衡向Th1方向滑动,通过其免疫调节作用,所述副干酪乳杆菌Probio‑37对过敏性鼻炎具有良好的缓解作用。所述副干酪乳杆菌Probio‑37可以广泛应用于其他食品及生物制品领域。
Description
技术领域
本申请涉及生物制剂领域,尤其涉及一种用于缓解过敏性鼻炎的副干酪乳杆菌及其应用。
背景技术
副干酪乳杆菌与人类、动物的生活关系密切,是一种常见于奶油、肉类及许多蔬菜、发酵制品和饲料中的乳酸菌,能通过胃并定植与肠道中发挥有益作用,目前,研究报道较多的应用领域包括食品、饲料、医疗保健、化妆品、美容、水产养殖、益生菌制剂等,当食用足够数量的细菌时,可能会给宿主带来一定的健康益处。
过敏性鼻炎(AR)属慢性上呼吸道炎疾病,是机体对某些变应原敏感性增高而发生在鼻腔粘膜的***反应性疾病,为临床常见的多发病之一。过敏性疾病虽然是非致命性疾病,但却严重影响患者的生活质量,给日常生活带来情感上、功能上和心理上的负面影响。根据流行病学的调查结果推算,过敏性鼻炎在人群中的患病率已达25%左右,其临床表现为:打喷嚏,鼻漏和鼻充血等症状。过敏性鼻炎主要分为两大类:常年性过敏性鼻炎(PAR)和季节性性过敏性鼻炎(SAR)。常年性过敏性鼻炎的特征是全年出现过敏性鼻炎症状,常见的过敏原包括室内尘螨,猫和狗皮屑,霉菌和真菌。季节性过敏性鼻炎对环境花粉(草、树、杂草等)有症状反应,而鼻症状评分与特定花粉季节有关。目前过敏性鼻炎被归为I型过敏性疾病,通常以血清免疫球蛋白E(IgE)水平升高为特征,这主要是由Ⅱ型辅助性T(Th2)细胞介导的,这些细胞合成高水平的白介素(IL)-4,IL-5和IL-13,从而导致过敏原特异性IgE的产生以及肥大细胞释放组胺和白三烯。
发明内容
本申请的目的之一在于提供一株具有缓解过敏性鼻炎症状的副干酪乳杆菌;
本申请的目的之二在于提供副干酪乳杆菌Probio-37在缓解过敏性鼻炎症状上的应用;
进一步地,所述副干酪乳杆菌Probio-37在发酵剂或添加的发酵乳中的应用;
进一步的,所述副干酪乳杆菌Probio-37在制备食品、保健品或药品中的应用。
为达到上述目的,本申请通过以下技术方案来实现:
所述副干酪乳杆菌Probio-37该菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCC No.18638;分类名称为副干酪乳杆菌(Lactobacillusparacasei),保藏时间:2019年10月8日;保藏地址:中国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;该菌株的检测结果为存活。
所述副干酪乳杆菌Probio-37(Lactobacillus paracasei Probio-37)对胃肠消化液具有良好的耐受性,能够以活的状态进入人体肠道。
本申请所述的副干酪乳杆菌Probio-37(Lactobacillus paracasei Probio-37)具有如下生物学特性:革兰氏阳性菌,无鞭毛、不运动,不形成芽孢,兼性厌氧;菌体呈杆状,其菌落光滑且边缘完整,呈乳白色。最适生长温度为30℃~38℃;最适pH值为6.0~7.0。
一种用于缓解过敏性鼻炎的产品,所述产品包含所述副干酪乳杆菌、所述副干酪乳杆菌的传代培养后代或由所述副干酪乳杆菌衍生出的突变株。
所述产品中,所述副干酪乳杆菌的活菌数不低于1×106CFU/mL或1×106CFU/g。
可选的,所述产品为副干酪乳杆菌乳粉或发酵剂。
可选的,所述产品为固体饮料、乳饮料或压片糖果。
可选的,所述产品包括功能性药品或保健品。
可选的,所述产品为使用含有所述发酵剂生产得到的豆制品、乳制品或果蔬制品。
由以上技术方案可知,本申请提供了一株用于缓解过敏性鼻炎的副干酪乳杆菌及其应用,所述副干酪乳杆菌对胃肠消化液具有良好的耐受性,能够以活的状态进入人体肠道。所述副干酪乳杆菌能刺激机体Th1/Th2免疫平衡向Th1方向滑动,通过其免疫调节作用缓解过敏性鼻炎。通过所述所述副干酪乳杆菌在食品、益生菌固体饮料、压片糖果、功能性药物辅助材料等领域的应用,以对过敏性鼻炎患者的症状进行有效缓解。
具体实施方式
一种用于缓解过敏性鼻炎的副干酪乳杆菌:所述副干酪乳杆菌Probio-37该菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCC No.18638;分类名称为副干酪乳杆菌(Lactobacillus paracasei),保藏时间:2019年10月8日;保藏地址:中国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;该菌株的检测结果为存活。
本申请所述的副干酪乳杆菌Probio-37(Lactobacillus paracasei Probio-37)具有如下生物学特性:革兰氏阳性菌,无鞭毛、不运动,不形成芽孢,兼性厌氧;菌体呈杆状,其菌落光滑且边缘完整,呈乳白色。最适生长温度为30℃~38℃;最适pH值为6.0~7.0。
所述副干酪乳杆菌Probio-37对胃肠消化液具有良好的耐受性,能够以活的状态进入人体肠道。
所述副干酪乳杆菌Probio-37能刺激机体Th1/Th2免疫平衡向Th1方向滑动,通过其免疫调节作用缓解过敏性鼻炎。
本发明实施例中实验方法和条件如无特别说明均为常规方法。
实施例1
副干酪乳杆菌Probio-37对小鼠过敏性鼻炎的缓解作用实验设计
将实验小鼠随机分为3组,进行过敏性鼻炎模型处理,
更为具体的是,第Ⅰ组对照组,采食无菌生理盐水中。
第Ⅱ组过敏性鼻炎组,在第0和7天进行卵清蛋白腹腔注射致敏(20μg卵清蛋白),从14-56天开始卵清蛋白双鼻孔滴鼻进行鼻刺激(200μg卵清蛋白)。
第Ⅲ组益生菌组,每天通过向小鼠给药,在第0和7天进行卵清蛋白致敏(20μg卵清蛋白),从14-56天开始用卵清蛋白进行鼻刺激(200μg卵清蛋白),同时给予副干酪乳杆菌Probio-37剂量为1×109CFU/kg体重(BW)每天灌胃,直至8W实验结束。
实验从最后一次鼻腔攻击后立即开始的10分钟内直至56天记录打喷嚏的次数。如表1所示,与对照组相比,经卵清蛋白刺激后小鼠打喷嚏次数明显增多,但益生菌组打喷嚏的次数显著低于过敏性鼻炎组。对照组的OVA特异性IgE水平明显低于其他组,这表明OVA致敏成功。与过敏性鼻炎组相比,口服益生菌组缓解显着降低了OVA特异性IgE含量。副干酪乳杆菌Probio-37能够显著降低过敏性鼻炎小鼠打喷嚏的次数和致敏因子,缓建鼻炎症状。
表1打喷嚏频率统计(±SD)
注:OVA-IgE,卵清蛋白特异性免疫球蛋白E。
每组随机选取7只小鼠,ELISA法检测其血清中个免疫因子的含量,血清进行5倍稀释后进行ELISA检测,结果如表2所示。
表2血清免疫指标
肠道免疫***中,Th1和Th2细胞会影响IgE的产生。Th2细胞分泌IL-6,IL-10和IL-4,导致IgE产生增加。Th1细胞分泌IFN-γ抑制IgE的合成以及对IgE介导的过敏的发作。
血清免疫细胞因子检测发现,过敏性鼻炎组Th2细胞分泌的IL-6,IL-10和IL-4因子含量均显著高于其他2组,而摄入Probio-37的益生菌组该类因子显著低于过敏性鼻炎组。而Th1细胞分泌的IFN-γ没有显著差异。这表明副干酪乳杆菌Probio-37通过调节Th1/Th2平衡向Th2方向滑动而抑制过敏性鼻炎的发生。
更为具体的是,实验菌株制剂为副干酪乳杆菌Probio-37菌粉制剂,活菌含量2×1011CFU/g,由北京科拓恒通生物技术股份有限公司提供。实验动物小鼠采用6月龄C57BL/6J小鼠,由北京维通利华实验动物技术有限公司提供,用于过敏性鼻炎模型建立。实验主要试剂盒为美国Sigma公司卵清蛋白(OVA)、氢氧化销佐剂;美国eBioScience公司ELISA试剂盒。
本申请以小鼠为动物模型进行的防治过敏性鼻炎研究发现,副干酪乳杆菌Probio-37能够调节肠道肠道免疫***,从而降低VOA引起的过敏性鼻炎反应,缓解鼻炎症状,对过敏性鼻炎有预防和治愈的作用。
实施例2
一种用于缓解过敏性鼻炎的食品,所述食品为所述副干酪乳杆菌Probio-37(Lactobacillus paracasei Probio-37)与可食性材料的组合物;所述产品中,所述副干酪乳杆菌Probio-37的活菌数不低于1×106CFU/mL或1×106CFU/g。
进一步的,所述可食性材料可以为:流体乳品(牛乳、流体牛乳)、奶粉、冰淇淋、乳酪或干酪。
优选的,一种副干酪乳杆菌乳粉的制造方法,将副干酪乳杆菌Probio-37的脱脂乳培养液与经过杀菌浓酸的牛乳成分混匀,再进行真空冷冻升华干燥,这种方法制造的副干酪乳杆菌乳粉对缓解过敏性鼻炎具有优异的效果。
实施例3
一种副干酪乳杆菌冰淇淋的制造方法,用常规方法得到冰淇淋,在分装之前加入实施例2所述的副干酪乳杆菌乳粉,搅拌均匀,即可得到副干酪乳杆菌冰淇淋。所述副干酪乳杆菌冰淇淋中,所述副干酪乳杆菌的活菌数不低于1×106CFU/mL或1×106CFU/g。对缓解过敏性鼻炎具有良好的作用。
实施例4
所述产品包括固体饮料、乳饮料或压片糖果。
更为具体的是,一种副干酪乳杆菌乳饮料制造方法。
使用浓缩脱脂乳或脱脂乳粉强化的脱脂乳,将混合料在98℃温度下加热杀菌40分钟,冷却到35℃,然后在其中添加4%的所述副干酪乳杆菌发酵剂,在35℃温度条件下培养,使酸度达到2%,培养时间为24小时,其中所述副干酪乳杆菌的活菌数不低于1×106CFU/mL或1×106CFU/g。得到所述副干酪乳杆菌乳饮料,其对缓解过敏性鼻炎具有良好的作用。
实施例5
所述产品包括功能性药品或保健品。所述功能性药品或保健品为所述副干酪乳杆菌Probio-37(Lactobacillus paracasei Probio-37)与药用辅料的组合物;所述产品中,所述副干酪乳杆菌Probio-37的活菌数不低于1×106CFU/mL或1×106CFU/g。
所述药用辅料包含溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、填充剂、润滑剂、润湿剂、渗透压调节剂、稳定剂、助流剂、矫味剂、防腐剂、助悬剂、包衣材料、芳香剂、抗黏合剂、整合剂、渗透促进剂、pH值调节剂、缓冲剂、增塑剂、表面活性剂、发泡剂、消泡剂、增稠剂、包合剂、保湿剂、吸收剂、稀释剂、絮凝剂与反絮凝剂、助滤剂以及释放阻滞剂中的一种或几种。
所述附加剂包括微晶纤维素、羟丙基甲基纤维素以及精制卵磷脂中的一种或几种。
所述药品或保健品的剂型可以为颗粒剂、胶囊剂、片剂、丸剂或口服液。
更为具体的是,一种副干酪乳杆菌胶囊的制造方法,将所述副干酪乳杆菌菌种用液体培养液培养,收集和洗涤菌体,添加药用辅料,干燥制备成活性菌粉,用明胶制成囊材,用PVP作底衣,再用蜂蜡进行外层包衣,用0号胶囊包装,得到副干酪乳杆菌胶囊。所述副干酪乳杆菌胶囊中,所述副干酪乳杆菌Probio-37的活菌数不低于1×106CFU/mL或1×106CFU/g。
实施例6
进一步的,所述产品包括使用含有所述副干酪乳杆菌发酵剂生产得到的豆制品、乳制品或果蔬制品。
更为具体的,一种副干酪乳杆菌发酵果蔬汁的制造方法。
首先取胡萝卜50公斤,经过分选、清洗、去皮、切分、漂烫、榨汁打浆、配料、匀质以及灭菌步骤得到无菌果蔬汁。
然后将副干酪乳杆菌菌种活化,扩大培养,接种到灭菌蔬菜汁、发酵、取样检测合格、冷却降温和无菌灌装,得到副干酪乳杆菌发酵果蔬汁,其中所述副干酪乳杆菌Probio-37的活菌数不低于1×106CFU/mL或1×106CFU/g。对缓解过敏性鼻炎具有良好的作用。
本领域技术人员在考虑说明书及实践这里公开的发明后,将容易想到本发明的其它实施方案。本申请旨在涵盖本发明的任何变型、用途或者适应性变化,这些变型、用途或者适应性变化遵循本发明的一般性原理并包括本发明未公开的本技术领域中的公知常识或惯用技术手段。说明书和实施例仅被视为示例性的,本发明的真正范围和精神由下面的权利要求指出。
Claims (4)
1.一种用于缓解过敏性鼻炎的副干酪乳杆菌,其特征在于,所述副干酪乳杆菌Probio-37该菌株保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCCNo.18638;分类名称为副干酪乳杆菌(Lactobacillus paracasei),保藏时间:2019年10月8日;保藏地址:中国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;该菌株的检测结果为存活。
2.权利要求1所述的副干酪乳杆菌Probio-37在缓解过敏性鼻炎症状的应用。
3.根据权利要求2所述的应用,其特征在于,所述副干酪乳杆菌Probio-37在制备发酵剂或添加的发酵乳中的应用。
4.根据权利要求2所述的应用,其特征在于,所述副干酪乳杆菌Probio-37在制备食品、保健品或药品中的应用。
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CN104523762A (zh) * | 2014-12-10 | 2015-04-22 | 中国农业大学 | 具有抗过敏作用的副干酪乳杆菌的应用和功能食品组合物及其制备方法 |
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