CN103002901A - 约汉逊氏乳杆菌La1，长双歧杆菌NCC2705和免疫疾病 - Google Patents
约汉逊氏乳杆菌La1，长双歧杆菌NCC2705和免疫疾病 Download PDFInfo
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- CN103002901A CN103002901A CN2011800354104A CN201180035410A CN103002901A CN 103002901 A CN103002901 A CN 103002901A CN 2011800354104 A CN2011800354104 A CN 2011800354104A CN 201180035410 A CN201180035410 A CN 201180035410A CN 103002901 A CN103002901 A CN 103002901A
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Abstract
本发明总地涉及益生菌的领域，特别地涉及例如通过增强内源性抗微生物防御来预防和/或治疗炎性或感染性疾病。本发明的一个实施方案是包含约汉逊氏乳杆菌(La1，NCC 533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合的组合物。该组合物可用于治疗或预防与免疫***相关的疾病，包括感染。
Description
本发明总地涉及益生菌领域，特别涉及预防和/或治疗炎性和感染性疾病的领域，例如通过增强内源性抗微生物防御来预防和/或治疗。本发明的一个实施方案是包含约汉逊氏乳杆菌La1(La1，NCC533保藏号CNCMI-1225)和长双歧杆菌(NCC2705保藏号CNCM I-2618)的组合的组合物。此组合物可用于治疗或预防与免疫***相关的疾病，包括感染。
我们的环境受到各种潜在致病的微生物的污染。皮肤角质细胞，胃肠道、呼吸道、泌尿-生殖道内层的上皮细胞都提供了进行保护以抵抗微生物侵入身体内的物理屏障。
此外，这些上皮细胞通过产生并分泌抗微生物剂来限制细菌和其他微生物的进入，从而有助于宿主防御。这些抗微生物分子构成了先天免疫的基础防线的关键组成部分。
防卫素是人类最重要的抗微生物肽的类别之一。防卫素由肺、皮肤、口腔、泌尿生殖器、呼吸道和胃肠道的上皮细胞产生。在这些中，存在着B-防卫素家族，包括防卫素1(hBD1)和防卫素2(hBD2)。
HBD1在多种粘膜表面中表达，如口腔粘膜、唾液腺、胃、小肠、结肠、肝和胰腺。HBD2也存在于多种粘膜表面的上皮细胞中，包括胃肠道的粘膜表面。此外，这两种防卫素还存在于唾液和气道表面流体中(Cunliffe，R.N.和Mahida，Y.R.2004，J Leukoc.Biol.75：49-58)。
HBD2以非常低的水平存在于正常组织中，并且其表达受到细菌和促炎性细胞因子的上调。与hBD2相反，HBD1是组成型表达的，HBD1从未显示出受到细菌或炎症的组成型上调(Ou，G.等，2009，Scand.JImmunol69：150-161)。
公知益生菌能够加强多种肠道防御系：免疫排斥、免疫排除和免疫调节。还已知益生菌刺激对微生物病原体的非特异性宿主抵抗，并且因此有助于微生物病原体的根除。
然而，尽管这样，已经报道了hBD1的组成型表达不受益生细菌的影响(O’Neil，D.A.等，J Immunol163：6718-6724)，并且受到共生菌株(大肠杆菌(Escherichia coli)和致病菌株(鼠伤寒沙门氏菌(Salmonellatyphimurium))非常轻微的上调(Ou，G.等，2009，Scand.J Immunol69：150-161)。
希望可获得没有不想要的副作用的有效的天然组合物，所述组合物能够用于上调β-防卫素，特别地上调防卫素1(hBD1)和2(hBD2)使得增强内源性抗微生物防御，使其在治疗或预防与免疫***相关的疾病，包括感染中是有用的。
理想地，天然组合物应当含有目前公认的和消费者认同的用于赋予健康益处的益生菌培养物。
能够有效刺激β-防卫素的表达，特别地防卫素1(hBD1)和2(hBD2)的表达的益生菌将具有额外的和/或改善的健康益处，这允许益生菌例如在治疗或预防与免疫***相关的疾病中特别有效。
本发明人解决了这种需求。
因此，本发明的目的是改进现有技术，以及提供天然并且有效的组合物，该组合物允许预防和/或治疗炎性和感染疾病，特别是通过增强内源性抗微生物防御来预防和/或治疗，并且满足上述需求。
发明人惊讶地看到他们可以通过独立权利要求的主题来实现本发明的目的。从属权利要求进一步限定了本发明的优选实施方案。
本发明的主题通过施用含有微生物(例如非复制性微生物，诸如热处理过的微生物)的产品加强了哺乳动物内源性的抗微生物防御。
发明人描述了约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合对诱导抗微生物肽表达具有优秀的效果。
发现，例如，约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合有效地触发hBD1和hBD2mRNA表达上调。引人注目地，观察到的作用比基于单个地测试两株菌株所产生的结果的预期显著更高。
观察到两种益生菌的不同组合所引起的hBD1mRNA表达上调比通过单个测试两株菌株所诱导的作用都更高。
关于hBD2表达，能够显示出，例如，较之用仅使用单个菌株所获得的hBD2的诱导，具有高剂量的长双歧杆菌(NCC2705，保藏号CNCMI-2618)(90)和低剂量的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCMI-1225)(10)的两种益生菌的组合导致增加的hBD2诱导。
因此，发明人已发现约汉逊氏乳杆菌(La1，NCC533，保藏号CNCMI-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合协同作用于诱导抗微生物肽(例如hBD1和/或hBD2)的表达。
HBD1和hBD2展示出对抗宽谱细菌(包括大肠杆菌和绿脓假单胞菌(Pseudomonas aeruginosa)、幽门螺旋杆菌(H.pylori)(Nuding，S.等，2009，Microbes.Infect.11：384-393))以及酵母(如白色念珠菌(Candidaalbicans)(O’Neil，D.A.2003，Mol.Immunol40：445-450))和病毒(人免疫缺陷病毒)(Kota，S.等，2008，J.Biol.Chem283：22417-22429)的抗微生物活性。因此，这些抗微生物肽可以加强粘膜屏障，并且因此限制细菌粘附和入侵。
越来越多的证据表明防卫素的水平在某些病理生理状况中下降并且这是感染性和炎性疾病的发病机理和并发症的风险因素，感染性和炎性疾病如(Doss，M.等，2010，J Leukoc.Biol87：79-92；Rivas-Santiago，B等，2009，Infect.Immun.77：4690-4695)：
-在呼吸道中：
囊性纤维化病、反应性气道疾病、肺部感染和抽烟、哮喘、肺炎、鼻炎、耳炎、窦炎、结核
-在胃肠道中：
克隆氏病(结肠和回肠)、溃疡性结肠炎、腹部疾病、肠不成熟、由幽门螺旋杆菌感染诱导的胃炎和胃溃疡、感染性腹泻、坏死性小肠结肠炎、抗生素相关的腹泻。
-在泌尿生殖道中：
细菌性***病、HIV、尿道感染
-在皮肤中：
特应性皮炎、慢性溃疡、癌症、特应性湿疹、烧伤
-在口腔中：
HIV病人、扁桃腺炎、龈炎、龋齿
-眼睛中的角膜炎
在此所呈现的结果表明约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合具有令人惊讶的强的增强基于协同作用的内源性抗微生物防御的能力，并且因此在SIBO(小肠细菌生长过度)、炎性和感染性疾病的预防和治疗中更有效。
因此本发明的一个实施方案涉及包含约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合的组合物。
可使用任意比例的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCMI-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。全部组合均比仅使用相应的量的单个菌株更有效。
然而，发现如果使用某些特别比例的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)，则例如在治疗或预防与免疫***相关的疾病中，约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合特别有效，例如通过增强内源性β-防卫素表达。
因此，本发明的组合物可包含比例在约10∶90至5∶95，优选地约50∶50至5∶95，例如约20∶80至5∶95的范围内的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。
例如，本发明的组合物可包含比例在约10∶90的范围内的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。
可基于克隆形成单位(cfu)的数目计算比例。
本发明的组合物可以含有足以至少部分地治疗与免疫***相关的疾病和/或其并发症的量的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCMI-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。将足以实现此的量定义为“治疗有效剂量”。处于该目的，有效的量将取决于本领域技术人员已知的多种因素，如疾病的严重程度、消费者的体重和一般健康状况以及食物基质的影响。
在预防性应用中，将根据本发明的组合物以足以至少部分地降低产生所述疾病风险的量施用于易患免疫***相关疾病的消费者或另外处于免疫***相关疾病风险中的消费者。将此类量定义为“预防有效剂量”。再次，精确的量取决于病人的多种特定因素，如病人的健康状况和体重，以及食物基质的影响。
本领域技术人员将能够适当地调整治疗有效剂量和/或预防有效剂量。
通常，本发明的组合物含有治疗有效剂量和/或预防有效剂量的La1。
通常，治疗有效剂量和/或预防有剂效量在每日剂量约0.005mg-1000mg约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的范围内。
就数量而言，约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)可以以对应于104至1012等价cfu/g干组合物的组合的量存在于组合物中。例如，根据本发明的组合物可以含有对应于每日剂量约104至1012cfu的量的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCMI-2618)。
本发明的组合物可以含有每日剂量约0.005mg-1000mg的约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。
本发明的组合物可以是任何种类的组合物。例如，可以口服、通过吸入、肠内、肠胃外(皮下或肌内)、局部或眼睛，直肠内和***内施用组合物。
因此，本发明的组合物可以选自食品组合物、食品产品(包括宠物食品)、饮料、完全营养配方、营养补充剂、营养制品、食品添加剂、药物组合物、化妆品组合物、局部组合物和药物。
本发明的组合物可用于治疗或预防与免疫***相关的疾病，例如感染。
根据本发明可通过增加内源性β-防卫素的表达(例如hBD1和/或hBD2表达)来治疗或预防与免疫***相关的疾病。
本发明的化合物也可用于治疗或预防与降低的β-防卫素表达(例如hBD1和/或hBD2表达)相关的疾病，举例而言，例如微生物感染。
本发明也涉及约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合在制备用于治疗或预防与免疫***相关的疾病的组合物中的用途。
与免疫***相关的疾病选自感染，特别是细菌、病毒、真菌和/或寄生虫感染；炎症；吞噬细胞缺乏；上皮屏障缺陷或免疫***不成熟、SIBO及其组合。
在本发明的一个实施方案中，本发明的组合物可以用于治疗或预防微生物感染，如病毒、真菌和/或寄生虫感染。
与免疫***相关的疾病还可以选自与降低水平的β-防卫素，特别是hBD1和/或hBD2相关的疾病。此类疾病可以选自囊性纤维化病、反应性气道疾病、抽烟引起的肺部感染、哮喘、肺炎、鼻炎、耳炎、窦炎、结核、克隆氏病(结肠和回肠)、溃疡性结肠炎、肠不成熟、上皮屏障缺陷或免疫***不成熟、SIBO、由幽门螺旋杆菌感染诱导的胃炎和胃溃疡、感染性腹泻、坏死性小肠结肠炎、抗生素相关的腹泻、细菌性***病、HIV、尿道感染、特应性皮炎、慢性溃疡、癌症、特应性湿疹、烧伤、扁桃腺炎、龈炎、龋齿、眼睛中的角膜炎，及其组合。
本发明的组合物可以用于增强内源性抗微生物防御。例如，这可以通过增强内源性hBD1，和/或hBD2表达来实现。
可添加益生元。益生元可支持益生菌的生长。此外，益生元还可与组合物中存在的活益生细菌协同作用。
本领域技术人员将理解他们可以自由地组合在此所述的本发明的所有特征，而没有脱离所公开的本发明的范围。特别地，对于本发明的组合物所述的特征可以应用至本发明的用途，反之亦然。
从以下的实施例和附图将清楚本发明的其他优点和特征。
图1显示了长双歧杆菌NCC2705(保藏号CNCM I-2618)与约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)的组合对hBD1mRNA表达的作用。将T84细胞与含有多种浓度的两株菌株的多种混合物孵育4h。通过实时PCR分析hBD1的基因表达。数据表示归一化至未受刺激的细胞的基础表达的平均值±sem。
图2显示出长双歧杆菌NCC2705(保藏号CNCM I-2618)与约汉逊氏乳杆菌(La1，NCC533，保藏号CNCM I-1225)的组合对hBD2mRNA表达的作用。将T84细胞与含有多种浓度的两株菌株的多种混合物孵育4h。通过实时PCR分析hBD2的基因表达。数据表示归一化至未受刺激的细胞的基础表达的平均值±sem。
实施例：
实验方案：
使用来自30-40代的T84细胞，并且在含有5％胎牛血清(FCS)(Amined BioConcept)和2mM谷胺酰胺的Dulbecco改良必需培养基/F-12(Sigma D6421)中培养。以2×106细胞/孔的浓度将细胞接种于6-孔培养平板中，并且在37℃下在5％CO2-95％空气气氛中生长成单层。用无血清和无抗生素的培养基将汇合后生长至1周的细胞培养至少12H。该步骤对于消除血清诱导的防卫素表达以及防止抗生素对益生菌和细胞免疫应答的任何影响是必需的。用益生菌将细胞再培养4H。在培养时间结束时，根据供应商的实验方案，用PBS洗涤细胞并且用TriPureTM分离试剂收获。通过定量PCR来评价如此处理的细胞中的人hBD1和hBD2基因表达。
该实验中所用的细菌菌株是长双歧杆菌(NCC2705保藏号CNCMI-2618)和约汉逊氏乳杆菌(La1，NCC533保藏号CNCM I-1225)。
结果：
由于长双歧杆菌(NCC2705保藏号CNCM I-2618)和约汉逊氏乳杆菌(La1，NCC533保藏号CNCM I-1225)分别特异性地上调hBD2和hBD1的mRNA表达，发明人研究了这两株菌株的组合制剂对肠抗微生物防御的作用。我们观察到这两种益生菌的不同的组合引起hBD1mRNA表达上调，所述表达上调都高于单个地测试由两株菌株诱导的作用(图1)。
关于hBD2表达，数据揭示，例如，较之用仅使用单个菌株所获得的hBD2的诱导，具有高剂量长双歧杆菌(NCC2705)(90)和低剂量La1(NCC533)(10)的两株益生菌的组合导致增加的hBD2诱导(图2)。
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仅用于国际局
Claims (11)
1.包含约汉逊氏乳杆菌(La1，NCC 533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)的组合的组合物。
2.根据权利要求1的组合物，所述组合物包含比例在10∶90至5∶95，优选地50∶50至5∶95，例如20∶80至5∶95范围内的约汉逊氏乳杆菌(La1，NCC 533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCMI-2618)。
3.根据前述权利要求中任一项的组合物，其中组合物含有含量对应于每日剂量约104至1012cfu的约汉逊氏乳杆菌(La1，NCC 533，保藏号CNCM I-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。
4.根据前述权利要求中任一项的组合物，其中组合物含有每日剂量约0.005mg-1000mg的约汉逊氏乳杆菌(La1，NCC 533，保藏号CNCMI-1225)和长双歧杆菌NCC2705(保藏号CNCM I-2618)。
5.)根据前述权利要求中任一项的组合物，其中组合物选自食品组合物、包括宠物食品的食品、饮料、完全营养配方、营养补充物、营养制品、食品添加剂、药物组合物、化妆品组合物、局部组合物和药物。
6.根据前述权利要求中任一项的组合物，所述组合物用于治疗或预防免疫***相关的疾病，包括感染。
7.根据权利要求1的组合物，其中与免疫***相关的疾病选自感染，特别是细菌、病毒、真菌和/或寄生虫感染；炎症；嗜菌细胞缺乏；上皮屏障缺陷或免疫***不成熟、SIBO及其组合。
8.根据权利要求6-7的组合物，其中与免疫***相关的疾病选自与降低水平的防卫素，特别是hBD1和hBD2相关的疾病。
9.根据权利要求8的组合物，其中与降低水平的防卫素相关的疾病选自囊性纤维化病、反应性气道疾病、肺部感染和抽烟、哮喘、肺炎、鼻炎、耳炎、窦炎、结核、克隆氏病(结肠和回肠)、溃疡性结肠炎、肠不成熟、由幽门螺旋杆菌感染诱导的胃炎和胃溃疡、感染性腹泻、坏死性小肠结肠炎、抗生素相关的腹泻、细菌性***病、HIV、尿道感染、特应性皮炎、慢性溃疡、癌症、特应性湿疹、烧伤、扁桃腺炎、龈炎、龋齿、眼睛中的角膜炎。
10.根据前述权利要求中任一项的组合物，所述组合物用于增强内源性抗微生物防御。
11.根据前述权利要求中任一项的组合物，所述组合物用于增强内源性hBD1和/或hBD2表达。
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| EP20100166526 EP2397145A1 (en) | 2010-06-18 | 2010-06-18 | L. johnsonii La1, B. longum NCC2705 and immune disorders |
| PCT/EP2011/060099 WO2011157816A1 (en) | 2010-06-18 | 2011-06-17 | L. johnsonii la1, b. longum ncc2705 and immune disorders |
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| JP (1) | JP5948323B2 (zh) |
| CN (1) | CN103002901B (zh) |
| ES (1) | ES2554693T3 (zh) |
| PT (1) | PT2582377E (zh) |
| WO (1) | WO2011157816A1 (zh) |
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| CN118028182A (zh) * | 2024-04-12 | 2024-05-14 | 四川厌氧生物科技有限责任公司 | 一种改善口腔健康的约翰逊氏乳杆菌及其用途 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104120093A (zh) * | 2013-04-26 | 2014-10-29 | 北京中天神舟航天食品技术研究院 | 长双歧杆菌及其应用以及功能食品组合物及其制备方法 |
| CN104120093B (zh) * | 2013-04-26 | 2016-12-28 | 中国农业大学 | 长双歧杆菌及其应用以及功能食品组合物及其制备方法 |
| CN105960244A (zh) * | 2013-12-10 | 2016-09-21 | 株式会社明治 | 抗菌肽诱导剂 |
| CN109652349A (zh) * | 2019-02-25 | 2019-04-19 | 江南大学 | 一株能够缓解特应性皮炎的长双歧杆菌及其应用 |
| CN114225024A (zh) * | 2021-12-29 | 2022-03-25 | 北京创世客生物技术有限公司 | 一种用于治疗疾病的益生菌药物或保健品 |
| CN114225024B (zh) * | 2021-12-29 | 2022-08-19 | 山东健袖生态科技有限公司 | 一种用于治疗疾病的益生菌药物 |
| CN118028182A (zh) * | 2024-04-12 | 2024-05-14 | 四川厌氧生物科技有限责任公司 | 一种改善口腔健康的约翰逊氏乳杆菌及其用途 |
Also Published As
| Publication number | Publication date |
|---|---|
| US9937212B2 (en) | 2018-04-10 |
| PT2582377E (pt) | 2015-11-30 |
| EP2397145A1 (en) | 2011-12-21 |
| JP2013530177A (ja) | 2013-07-25 |
| ES2554693T3 (es) | 2015-12-22 |
| US20130089524A1 (en) | 2013-04-11 |
| EP2582377B1 (en) | 2015-09-09 |
| JP5948323B2 (ja) | 2016-07-06 |
| CN103002901B (zh) | 2016-06-29 |
| EP2582377A1 (en) | 2013-04-24 |
| WO2011157816A1 (en) | 2011-12-22 |
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