CN112280706A - 一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌及应用 - Google Patents
一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌及应用 Download PDFInfo
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Abstract
本发明涉及一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌及应用,所述植物乳杆菌的菌株编号为PP12,分类命名为植物乳杆菌Lactobacillus plantarum,保藏号为:CGMCC No 8871,保藏日期为2014年2月27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。本发明利用抗过敏的植物乳杆菌PP12,可以解决使用药物预防或治疗过敏反应时副作用大的问题。此外,来源于发酵食品的植物乳杆菌PP12安全有效,在发挥抗过敏功能的同时还可以调节肠道菌群平衡,预防其他免疫系统疾病。本发明为治疗过敏性疾病提供了新方向,可应用于食品、保健品和药品领域,具有非常好的产业前景。
Description
技术领域
本发明涉及一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌及应用,属于微生物技术应用领域。
背景技术
过敏反应是指已产生免疫的机体在再次接受相同抗原刺激时所发生的组织损伤或功能紊乱的反应。过敏的成因是身体内如果第二型T辅助细胞 (TH2)的活性过高、过盛就能制造较多的过敏抗体IgE,就会出现过敏现象。人体的免疫细胞可分为Th1和Th2,当过敏原与身体接触时,免疫耐受功能异常,导致Th2细胞过度活化,过敏抗体IgE分泌增加,致使下游免疫细胞产生大量细胞激素如组胺、白三烯等,从而出现过敏现象如过敏性鼻炎、哮喘、湿疹、荨麻疹、过敏性结膜炎等症状,但如果肠道内有足够的抗过敏益生菌,就能够刺激肠道壁上的树突状细胞分泌白介素-I2(IL-I2),就能够针对过敏上游源头透过对免疫辅助T细胞的调控,通过增进Th1型免疫反应,降低IgE从而降低反应过度的Th2免疫反应的方法达到有效缓解过敏反应,改善过敏体质。
过敏性疾病,如过敏性鼻炎、异位性皮炎、过敏性哮喘和食物过敏,已在许多国家变得越来越普遍。这些疾病不仅影响个人的生活品质,而且成为社会医疗负担。
植物乳杆菌是肠胃道中最常见的益生菌,是一种安全性极高的微生物,不仅在诸多发酵食品,而且在厌氧性食品或食物等物质中所发现,广泛存在于自然界。植物乳杆菌具有抑制有害菌的生长、调节免疫反应、清除过氧化物及降低胆固醇等能力。此外,植物乳杆菌含有宜人的香味物质,并能产生乳酸或细菌素(bacter iocin)等抗菌性物质,可以用于食品的制备。
发明内容
本发明所要解决的技术问题是提供一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌及应用,可以制备预防和/或治疗过敏性疾病的产品,解决上述技术问题。
本发明解决上述技术问题的技术方案如下:一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌,所述植物乳杆菌的菌株编号为PP12,分类命名为植物乳杆菌Lactobacillus plantarum,保藏号为:CGMCC No 8871,保藏日期为2014年2月27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,保藏单位地址:北京市朝阳区北辰西路1号院3号。
本发明还涉及一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌在制备预防和/或治疗过敏性疾病的产品中的应用。
在上述技术方案的基础上,本发明还可以做如下改进。
进一步,所述过敏性疾病选自过敏性鼻炎、异位性皮炎、过敏性哮喘、食物过敏中的一种。
进一步,所述产品包括食品、保健品或药品。
进一步,所述产品为抗过敏饮料。
进一步,所述抗过敏饮料由发酵原液与植物乳杆菌PP12混合发酵后加入添加剂制得。
进一步,以重量份计,所述发酵原液包括生活饮用水85.5-93.9份、脱脂奶粉2.2-5.5份、葡萄糖1.5-2.5份,所述添加剂包括低聚糖0.05-3份、果葡糖浆2.0-3.0份、果胶0.3-0.5份、甜味剂0.01-0.04份、酸度调节剂0.01-0.03份、食用香精0.003-0.005份,在100kg发酵原液中添加植物乳杆菌PP12 5-10IU。
本发明的有益效果是:本发明利用抗过敏的植物乳杆菌PP12,可以解决使用药物预防或治疗过敏反应时副作用大的问题。此外,来源于发酵食品的植物乳杆菌PP12安全有效,在发挥抗过敏功能的同时还可以调节肠道菌群平衡,预防其他免疫系统疾病。本发明为治疗过敏性疾病提供了新方向,可应用于食品、保健品和药品领域,具有非常好的产业前景。
具体实施方式
以下对本发明的原理和特征进行描述,所举实例只用于解释本发明,并非用于限定本发明的范围。
本发明所用的试剂、培养基组成以及实验学方法等在没有特别说明的情况下,均为本领域常规使用的试剂、培养基组成以及实验学方法。在没有特别说明的情况下,以下所测得的数值均是在实验条件下测定至少8个平行样品所得的平均值。
实施例1植物乳杆菌PP12的分离筛选
本发明植物乳杆菌PP12是从发酵泡菜中筛选出来的最优菌株,其具体筛选方法如下:
(一)菌种的分离筛选
取10g的泡菜来制备泡菜破碎液。泡菜破碎条件为使用搅拌机以 2000rpm充分破碎2分钟,之后,利用106倍的灭菌的蒸馏水稀释并涂抹于MRS 培养基后,培养24小时,收集30个菌落。
(二)筛选抗过敏的菌种
分别取30个菌落接种于MRS培养液中,在30℃下培养21小时,利用离心(1500grpm,10分钟)收集,并以无菌PBS洗涤两次,然后再悬浮于PBS中,至最终浓度为1010CFU/ml,得到各个菌落的活菌体,并储存在-20℃直至使用。
取四周龄雌性BALB/c品系小鼠,将动物房维持在12:12小时光照-黑暗的周期、温度为25±2℃、以及湿度55±15%。以标准实验室饮食喂食小鼠, 适应两周后,进行OVA致敏及细菌喂养。
将6周龄小鼠以0VA致敏及诱发,以建立经OVA引起的呼吸道过敏 BALB/c小鼠模型。将小鼠分组进行不同的处理34天。对照组和实验组均使用不锈钢喂食管经口给予PBS,实验组每天还分别给予109CFU不同的活菌体,并在第1天及第14天以含有50μg的OVA的100μL的Al(OH)3进行腹腔注射。于第28天、第29天及第30天,通过鼻内给药且以0VA(1%于PBS中,100 μL/小鼠,每天)或PBS诱发小鼠。在第32天,测定小鼠的AHR。在第34天, 牺牲所有小鼠,使用眼窝静脉采血收集血液,并通过离心分离(2000rpm离心 10分钟)以制备血清。于免疫球蛋白分析之前,将血清保存在-20C。
在研究期间,每天测定小鼠体重。各组间的食物摄入、喂食效率或体重变化并无显著差异。
于第7天,0VA致敏小鼠的血清总IgE升高,并持续增加至第34天。于第 34天,相较于对照组,接受某些活菌体显示血清总IgE显著下降(P<0.01)。其中选取血清总IgE最低的菌株,并进行菌种鉴定。
利用形态特征、培养性状和生理生化特征等微生物学特性以及16SrDNA 等分子生物学手段对该菌株鉴定为植物乳杆菌,将其进行菌种保藏,菌株编号为PP12,分类命名为植物乳杆菌Lactobacillus plantarum,保藏号为: CGMCC No 8871,保藏日期为2014年2月27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。
实施例2:利用植物乳杆菌PP12制备抗过敏饮料
方案1:一种抗过敏饮料的制备方法,由发酵原液与植物乳杆菌PP12 混合发酵后加入添加剂制得。以重量份计,所述发酵原液包括生活饮用水86 份、脱脂奶粉2份、葡萄糖1.5份,所述添加剂包括低聚糖0.05份、果葡糖浆2份、果胶0.3份、甜味剂0.01份、酸度调节剂0.01份、食用香精0.003 份,在100kg发酵原液中添加植物乳杆菌PP12 5IU。
包括以下步骤:
(1)脱脂奶粉复原:用55℃的生活饮用水将脱脂奶粉复原,加入葡萄糖, 经巴氏杀菌后直接使用或升温至90℃以上进行褐变处理得发酵原液;
(2)发酵:将发酵原液温度调至适宜菌种生长的37℃,无菌操作加入植物乳杆菌PP12进行发酵,发酵时间为15hh;
(3)调配:向发酵液这加入低聚糖、果葡糖浆、果胶、甜味剂、酸度调节剂、食用香精;
(4)均质:压力在18MPa下进行均质处理;
(5)灌裝;
(6)包装入库。
方案2:一种抗过敏饮料的制备方法,由发酵原液与植物乳杆菌PP12 混合发酵后加入添加剂制得。以重量份计,所述发酵原液包括生活饮用水94 份、脱脂奶粉5.5份、葡萄糖2.5份,所述添加剂包括低聚糖2份、果葡糖浆3份、果胶0.5份、甜味剂0.03份、酸度调节剂0.03份、食用香精0.005 份,在100kg发酵原液中添加植物乳杆菌PP12 10IU。
包括以下步骤:
(1)脱脂奶粉复原:用65℃的生活饮用水将脱脂奶粉复原,加入葡萄糖, 经巴氏杀菌后直接使用或升温至90℃以上进行褐变处理得发酵原液;
(2)发酵:将发酵原液温度调至适宜菌种生长的38℃,无菌操作加入植物乳杆菌PP12进行发酵,发酵时间为72h;
(3)调配:向发酵液这加入低聚糖、果葡糖浆、果胶、甜味剂、酸度调节剂、食用香精;
(4)均质:压力在25MPa下进行均质处理;
(5)灌裝;
(6)包装入库。
方案3:一种抗过敏饮料的制备方法,由发酵原液与植物乳杆菌PP12 混合发酵后加入添加剂制得。以重量份计,所述发酵原液包括生活饮用水90 份、脱脂奶粉4份、葡萄糖2份,所述添加剂包括低聚糖1份、果葡糖浆2.5 份、果胶0.4份、甜味剂0.02份、酸度调节剂0.02份、食用香精0.004份,在100kg发酵原液中添加植物乳杆菌PP12 7.5IU。
包括以下步骤:
(1)脱脂奶粉复原:用60℃的生活饮用水将脱脂奶粉复原,加入葡萄糖, 经巴氏杀菌后直接使用或升温至90℃以上进行褐变处理得发酵原液;
(2)发酵:将发酵原液温度调至适宜菌种生长的37℃,无菌操作加入植物乳杆菌PP12进行发酵,发酵时间为48h;
(3)调配:向发酵液这加入低聚糖、果葡糖浆、果胶、甜味剂、酸度调节剂、食用香精;
(4)均质:压力在22MPa下进行均质处理;
(5)灌裝;
(6)包装入库。
本发明的植物乳杆菌PP12具有抗过敏的功效,低聚糖可以通过增殖肠道中的双歧杆菌等有益菌,增强机体免疫力,更好分解奶粉中的乳糖,减缓乳糖耐症。通过大量实验所得的本发明的技术方案,经验证,发现该饮料不仅能有效的增强机体免疫力,而且可以调整IgE、改善免疫耐受,抗敏效果好,值得推广。
尽管上面已经示出和描述了本发明的实施例,可以理解的是,上述实施例是示例性的,不能理解为对本发明的限制,本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
Claims (7)
1.一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌,其特征在于,所述植物乳杆菌的菌株编号为PP12,分类命名为植物乳杆菌Lactobacillus plantarum,保藏号为:CGMCCNo 8871,保藏日期为2014年2月27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。
2.一种根据权利要求1所述一种可调整IgE、改善免疫耐受、抗过敏的植物乳杆菌在制备预防和/或治疗过敏性疾病的产品中的应用。
3.根据权利要求2所述的应用,其特征在于,所述过敏性疾病选自过敏性鼻炎、异位性皮炎、过敏性哮喘、食物过敏中的一种。
4.根据权利要求2所述的应用,其特征在于,所述产品包括食品、保健品或药品。
5.根据权利要求4所述的应用,其特征在于,所述产品为抗过敏饮料。
6.根据权利要求5所述的应用,其特征在于,所述抗过敏饮料由发酵原液与植物乳杆菌PP12混合发酵后加入添加剂制得。
7.根据权利要求6所述的应用,其特征在于,以重量份计,所述发酵原液包括生活饮用水86-94份、脱脂奶粉2-5.5份、葡萄糖1.5-2.5份,所述添加剂包括低聚糖0.05-2份、果葡糖浆2-3份、果胶0.3-0.5份、甜味剂0.01-0.03份、酸度调节剂0.01-0.03份、食用香精0.003-0.005份,在100kg发酵原液中添加植物乳杆菌PP12 5-10IU。
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