CN104870632A - 鼠李糖乳杆菌菌株用于降低体重增加和/或胰岛素耐受性 - Google Patents
鼠李糖乳杆菌菌株用于降低体重增加和/或胰岛素耐受性 Download PDFInfo
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Abstract
本申请提供了鼠李糖乳杆菌菌株CNCM I-3690用于降低饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性以及治疗受试者中由此产生的紊乱如超重、肥胖和与肥胖相关的紊乱的用途。
Description
本发明涉及益生菌领域，且更具体地涉及其用于调节体重增加和胰岛素耐受性的用途。
过去25年超重和肥胖在全世界的许多地方的流行大幅增加。身体质量指数(BMI)大于或等于25被认为是超重，身体质量指数大于或等于30被定义为肥胖。
自1980年，全球的肥胖人口的数量已增加超过一倍。在2008年，超过14亿的成人(20岁及以上)超重。这其中，有超过2亿男性和近3亿女性肥胖。
肥胖通常与胰岛素耐受性相关(即细胞不再能够适当地响应于胰岛素的状况)，导致包括代谢综合症的多数疾病，例如高血压、II型糖尿病、心血管疾病以及肝脏疾病。
虽然个体间的脂肪积累以及体重的差异可能与各种因素相关，例如遗传背景、健康状况、医学治疗或年龄，但普遍认为目前频率的肥胖和胰岛素耐受性的主要原因之一是日常生活中运动量减少以及富含高脂肪和高糖食物的西式饮食的结合。
同样，近年来的大量研究表明，肥胖和与肥胖相关的代谢紊乱与肠道菌群组成的变化相关(参见LEY,Curr Opin Gastroenterol.26,5-11,2010)，并且肠道菌群可影响宿主代谢，以及提高从饮食获得的能量(MARIK,FrontEndocrinol(Lausanne),3,87,2012)。
因此，曾有人建议将肠道菌群的调节作为改善代谢综合症的靶标。这可通过使用益生元、益生菌或合生元而实现，益生元、益生菌或合生元可有助于降低肥胖和与肥胖相关的代谢紊乱，包括胰岛素耐受性(MALLAPPA等人，Indian J Endocrinol Metab,16,20-7,2012；DELZENNE等人，Nat RevEndocrinol,7,639-46,2011)。
已报道某些益生菌菌株降低了脂肪积累和/或与肥胖相关的代谢紊乱。LEE等人(Biochim Biophys Acta,1761,736-44,2006)已经表明，产生结合性亚油酸的鼠李糖乳杆菌菌株PL60(Lactobacillus rhamnosus strain PL60)在饮食诱导的肥胖小鼠中具有抗肥胖作用；加氏乳杆菌SBT2055(Lactobacillusgasseri SBT2055)已显示在成年人中降低腹部肥胖和体重(KADOOKA等人，Eur J Clin Nutr,64,636-43,2010)，并且加氏乳杆菌BNR17(Lactobacillusgasseri BNR17)已显示在喂食高碳水化合物饮食的大鼠中减少体重增加(KANG等人，J Microbiol,48,712-4,2010)。ANDREASEN等人(Br J Nutr,104,1831-8,2010)报道了施用嗜酸乳杆菌NCFM(Lactobacillus acidophilusNCFM)对胰岛素耐受性的改善。植物乳杆菌第14号菌株(Lactobacillusacidophilus strain No.14)显示降低了喂食高脂肪饮食的小鼠中脂肪细胞的大小(TAKEMURA等人，Exp Biol Med(Maywood),235,849-56,2010)。ARONSSON等人(PLoS One,5,2010)报道，副干酪乳杆菌副干酪亚种菌株F19(Lactobacillus paracasei ssp paracasei strain F19)能够通过提高血管生成素样4蛋白(ANGPTL4)的表达来降低脂肪储存。MA等人(J Hepatol,49,821-30,2008)报道，益生菌VSL#3通过增加肝NKT细胞改善了高脂肪饮食诱导的肝脂肪变性和胰岛素耐受性。JI等人(Benef Microbes,3,13-22,2012)报道了鼠李糖乳杆菌GG(Lactobacillus rhamnosus GG)和沙克乳杆菌NR28(Lactobacillus sakei NR28)对鼠类菌群的调整，其伴随有抗肥胖作用。AN等人(Lipids Health Dis,10,116,2011)描述了在高脂肪饮食诱导的肥胖大鼠中双歧杆菌菌株混合物(假小链双歧杆菌(B.pseudocatenulatum)SPM1204、长双歧杆菌(B.longum)SPM 1205和长双歧杆菌SPM 1207)的抗肥胖和降脂质作用。PCT申请WO 2007/043933提出了干酪乳杆菌F19(Lactobacillus casei F19)、嗜酸乳杆菌NCFB 1748(Lactobacillus acidophilusNCFB 1748)或乳双歧杆菌Bb12(Bifidobacterium lactis Bb12)用于降低食物摄入和脂肪沉积以及预防或治疗肥胖和胰岛素不敏感性的用途。AMAR等人(AMAR等人，EMBO Mol Med,3,559-72,2011)显示，施用益生菌株乳双歧杆菌B420给糖尿病小鼠提高了空腹血糖，并恢复葡萄糖转化速率到喂饲正常饮食的对照小鼠的水平。
这些不同益生菌的作用是菌株特异性的，并且似乎是由不同的机制所介导。因此，仍然需要能够用于控制超重和肥胖以及与之相关的代谢疾病的发展的其它益生菌菌株。
本发明人已经着手研究小鼠中益生菌对饮食诱导的肥胖和胰岛素耐受性的预防作用。他们发现，给饲喂高脂肪饮食(HFD)的小鼠以108细胞/天口服施用鼠李糖乳杆菌菌株CNCM 1-369012周显著地降低了体重增加、胰岛素耐受性。此外，通过454焦磷酸测序16S rRNA基因对肠道菌群的分析显示，鼠李糖乳杆菌菌株CNCM 1-3690与HF饮食相比改变了肠道菌群的结构。更具体的分析揭示，菌株CNCM 1-3690改变了不同运算分类单位(OUT)的相对丰度，但是大多数提高的OUT来自产生乳酸(脂)盐和醋酸(脂)盐的细菌。盲肠内含物的短链脂肪酸分析证实了与HFD相比，菌株CNCM1-3690显著地提高了肠道醋酸(脂)盐的量，这与通过454焦磷酸测序的微生物群分析一致。
菌株CNCM I-3690依据布达佩斯条约于2006年11月19日保藏于CNCM。该菌株在PCT申请WO 2009/122042中被公开，此申请报道了该菌株的抗微生物和免疫调节性质。该菌株还具有抗氧化性质，这在PCT申请WO 2011/083354中有描述。
因此，本发明涉及的主题是鼠李糖乳杆菌菌株CNCM I-3690或包含所述菌株的组合物用于降低受试者中饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性的用途。
“饮食诱导的体重增加”和“饮食诱导的胰岛素耐受性”文中定义为饮食摄入过量的脂肪(特别是不饱和脂肪)和/或单糖(尤其包括蔗糖和果糖)导致的体重增加和胰岛素耐受性。对于给定的受试者，过量地摄入饮食脂肪和/或单糖是指消耗的饮食脂肪和/或单糖的量大于满足所述受试者的生理需求并维持其能量平衡必需的量。可通过比较在接受治疗的受试者中观察到的体重增加或胰岛素耐受性与接受相同饮食并具有相同水平的身体活动但未接受治疗的相同受试者中观察到的的体重增加或胰岛素耐受性，评估所述治疗对降低饮食诱导的体重增加或胰岛素耐受性的作用
现有技术中已知评估受试者中的胰岛素耐受性的测试(参见例如FERRANNINl&MARI,J Hypertens.,16,895-906,1998)。可用本领域中已知的任何胰岛素耐受性测试，例如胰岛素耐受性的稳态模型评估(HOM-IR)，测定受试者的胰岛素耐受性水平。
本发明还包括鼠李糖乳杆菌菌株CNCM I-3690或包含所述菌株的组合物，用于治疗、预防或缓解由饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性所导致的病症。
由饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性所导致的病症的实例为超重、肥胖和相关的紊乱如2型糖尿病、非酒精性脂肪肝病(NAFLD)、高血压等。
本发明还提供了降低有需要的受试者中饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性的方法，其中，所述方法包括向所述受试者施用鼠李糖乳杆菌菌株CNCM I-3690或包含所述菌株的组合物。
鼠李糖乳杆菌菌株CNCM I-3690可以以活的或死的完整细菌的形式使用。或者，该菌株可以以细菌裂解物的形式或细菌级分(bacterial fraction)的形式使用
本发明中使用的组合物可以是任何适合于给药尤其是口服给药的形式。这包括例如固体、半固体、液体和粉末。由于易于施用，通常优选液体组合物，如作为饮料。
当细菌是活细菌的形式时，以组合物的每克干重计，所述组合物通常可以包含105至1013个菌落形成单位(cfu)，优选至少106cfu，更优选至少107cfu，还更优选至少108cfu，并且最优选至少109cfu。在液体组合物的情况下，这通常对应于104至1012个菌落形成单位(cfu)，优选至少105cfu，更优选至少106cfu，还更优选至少107cfu，并且最优选至少109cfu/ml。
本发明中使用的优选的组合物是营养组合物，包括食品并且尤其是乳产品。这些营养组合物还包括食品补充剂和功能性食品。“食品补充剂”是指由通常在食品中所使用的化合物所制成的产品，但其形式是片剂、粉剂、胶囊剂、饮剂或通常与营养物无关的任何其它形式，且对人的健康具有有益作用。“功能性食品”是也对人的健康具有有益作用的营养物(aliment)。具体地，食品补充剂和功能性食品针对疾病，例如针对慢性疾病，具有生理学作用——防护或治疗性的作用。
适合用在本发明中的组合物的其它实例有药物组合物。
除了菌株CNCM I-3690之外，本发明的组合物还可以包含益生性的或非益生性一个或多个其它的乳酸细菌的菌株，例如选自菌属乳杆菌(Lactobacillus)、乳球菌(Lactococcus)、链球菌(Streptococcus)和双歧杆菌(Bifidobacteria)的一个或多个细菌菌株。具体地，这个(这些)其它菌株可包括嗜热链球菌中的一个或多个菌株，和/或保加利亚乳杆菌中的一个或多个菌株。
可以由如下进一步描述的引用例示细菌菌株CNCM I-3690对脂质储备的作用的实施例，来更清楚地理解本发明。
附图说明
图1：对于四个组：NC、HFD、HFD+CNCM 1-3690、HFD+乳双歧杆菌B420(Danisco)的体重增加(A)、空腹血糖(B)、空腹胰岛素(C)、HOMA-IR(D)、OGTT(E)以及OGTT(F)的曲线下面积(AUC)。数据显示为平均值±S.E.M。当在SPSS中通过单侧-ANOVA然后Tukey事后检验时，与HFD组相比，**p<0.01，*p<0.05，当与NC组比较时，##p<0.01，#p<0.05。根据下面的式子计算HOMA-IR：空腹血糖(mmol/L)×空腹胰岛素(mU/L)/22.5。
图2：四个组每周的食物摄取。数据显示为两笼小鼠的平均值，因此未进行统计分析。
图3：四个组在动物实验的每个月的累积食物摄取。数据显示为两笼小鼠的平均值，因此未进行统计分析。
图4：四个组在12周的期间的累积食物摄取。数据显示为两笼小鼠的平均值，因此未进行统计分析。
实施例1：鼠李糖乳杆菌CNCM I-3690改善了高脂肪饮食诱导的肥胖和胰岛素耐受性。
材料和方法
C57BL/6J小鼠(雄性，12周龄)分成3个组(每组8只小鼠)，进行如下不同处理：
A组：高脂肪饮食(HFD)，包含34.9％的脂肪，5.24的kcal/g，来自Research Diets Inc.,New Brunswick,NJ；
B组：高脂饮食加上108CFU/小鼠/天的益生菌菌株鼠李糖乳杆菌CNCM1-3690(HFD+CNCM 1-3690)；
C组：高脂饮食加上108CFU/小鼠/天的益生菌菌株乳双歧杆菌B420(Danisco)(HFD+乳双歧杆菌B420)，先前报道降低了对与高脂肪饮食相关的代谢的副作用(AMAR等人，2011，上文有引用)，作为对照；
D组：标准饮食(NC)，包含4.3％的脂肪，3.85kcal/g，来自ResearchDiets Inc.,New Brunswick,NJ。
在动物实验前制备鼠李糖乳杆菌CNCM 1-3690和乳双歧杆菌B420悬浮液，储存在-80℃，并在通过经口喂养而施用到每只小鼠前1小时解冻。
持续治疗动物12周，在此期间每周测量每只小鼠的体重和每笼小鼠的食物摄入两次。通过使用代谢笼每月收集新鲜的粪便和尿样本一次，并立即储存在-80℃用于后面的分析。
通过反转录(RT)-qPCR量化在施用益生菌期间的第2周、第6周和第11周的小鼠排泄物中益生菌的量，并且结果证实它们可在肠中存活。
在实验的结束时，缺乏食物5个小时后，从眼窝血管丛收集血液，并通过在4℃以3000rpm离心15分钟而分离血清。通过颈脱位法处死全部的动物。在处死后切除附睾脂肪垫、肝脏和空肠、称重并立即保存在RNALater(Ambion)中。
在处死动物前进行口服葡萄糖耐量试验(OGTT)。在缺乏食物5个小时后，小鼠口服施用2.0g/kg体重的葡萄糖。从尾部取血样，以使用ACCU-Check葡萄糖仪(Roche Diagnostics，加拿大)测量施用葡萄糖前以及施用葡萄糖后15、30、60和120分钟的血糖水平。认为施用葡萄糖前的血糖水平为空腹葡萄糖(FBG)水平。通过ELISA分析(Mercodia，瑞典)测定空腹胰岛素(FINS)水平。根据下面的式子计算HOMA-IR：空腹血糖(mmol/L)×空腹胰岛素(mU/L)/22.5。
结果
图1中显示了结果：A：体重增加；B：空腹血糖水平(FBG)；C：空腹胰岛素水平(FINS)；D：胰岛素耐受性的动态平衡评估(HOMA-IR)指数；E：口服葡萄糖耐量试验(OGTT)曲线，F：OGTT的曲线下面积(AUC)。数据显示为平均值±S.E.M。当在SPSS中通过单侧-ANOVA然后Tukey事后检验时，与HFD组相比，**p<0.01，*p<0.05，与NC组相比，##p<0.01，#p<0.05。
与喂饲NC的小鼠相比，HFD组显示较高的体重增加(图1A)，提高的空腹血糖水平(图1B)，提高的空腹胰岛素水平(图1C)和提高的HOMA-IR水平(图1D)以及降低的葡糖糖耐受性(图1E、F)。向HFD饲料中补充两种益生菌菌株显著降低了体重增加(图1A)。虽然在HFD+益生菌组和HFD组之间空腹血糖(FBG)水平和空腹胰岛素(FINS)水平没有显著差异，但是鼠李糖乳杆菌CNCM 1-3690或乳双歧杆菌B420均降低了HOMA-IR指数，并且该降低在CNCM 1-3690的情况下是显著的(图1D)。这两种菌株还显著地减小了葡萄糖耐受性(图1E、F)，表明它们可改善胰岛素耐受性。
计算所述实验的12周中每周的每天每只小鼠的平均能量摄入(图2)。在全部的实验中，NC组的能量摄入最低，而HFD+益生菌组的能量摄入除了第7周以外几乎与HFD组相同。此外，计算了四个组的动物在3个月的期间的累积能量摄入(图3)和四个组的动物在12个周的期间的累积能量摄入(图4)。这表明观察到的益生菌处理组的体重降低不能归因于能量摄入的减少。
这些结果表明，所述两种益生菌菌株显著地改善了由HFD引起的肥胖和胰岛素耐受性，并且鼠李糖乳杆菌菌株CNCM 1-3690产生的改善作用至少相当于乳双歧杆菌B420产生的改善。
Claims (6)
1.鼠李糖乳杆菌(Lactobacillus rhamnosus)菌株CNCM I-3690，用于降低受试者中饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性。
2.权利要求1的用于所述用途的鼠李糖乳杆菌菌株CNCM I-3690，其中所述用途是用于治疗、防止或缓解由饮食诱导的体重增加和/或饮食诱导的胰岛素耐受性所导致的病症。
3.权利要求1或2中任一项的用于所述用途的鼠李糖乳杆菌菌株CNCMI-3690，其中所述病症选自超重、肥胖和与肥胖相关的紊乱。
4.权利要求1至3中任一项的用于所述用途的鼠李糖乳杆菌菌株CNCMI-3690，其中所述菌株包含于可口服施用的组合物中。
5.权利要求4的用于所述用途的鼠李糖乳杆菌菌株CNCM I-3690，其中所述组合物是食品或食品补充剂。
6.权利要求5的用于所述用途的鼠李糖乳杆菌菌株CNCM I-3690，其中所述食品是发酵的乳产品。
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| CN112672749A (zh) * | 2018-04-11 | 2021-04-16 | 韩国生命工学研究院 | 具有肥胖预防或治疗效果的新的长双歧杆菌菌株或鼠李糖乳杆菌菌株及其用途 |
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| CN112672749B (zh) * | 2018-04-11 | 2024-07-23 | 韩国生命工学研究院 | 具有肥胖预防或治疗效果的新的长双歧杆菌菌株或鼠李糖乳杆菌菌株及其用途 |
| CN111164201A (zh) * | 2018-07-02 | 2020-05-15 | 葡萄王生技股份有限公司 | 用于改善代谢症候群的新颖副干酪乳杆菌gks6、其培养基、培养方法、用途、医药组合物及可食用组合物 |
| CN111164201B (zh) * | 2018-07-02 | 2023-09-12 | 葡萄王生技股份有限公司 | 用于改善代谢症候群的副干酪乳杆菌gks6、其用途、医药组合物及可食用组合物 |
| CN109757730A (zh) * | 2019-01-31 | 2019-05-17 | 山东环亿生物科技有限公司 | 一种具有降血脂、血压以及血糖的组合物及其制备方法 |
| CN109757730B (zh) * | 2019-01-31 | 2021-06-04 | 山东环亿生物科技有限公司 | 一种具有降血脂、血压以及血糖的组合物及其制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| ES2675309T3 (es) | 2018-07-10 |
| US10022407B2 (en) | 2018-07-17 |
| MX2015005898A (es) | 2016-02-05 |
| JP2016501010A (ja) | 2016-01-18 |
| CA2891618A1 (en) | 2014-05-15 |
| US20150297646A1 (en) | 2015-10-22 |
| RU2015122748A (ru) | 2017-01-10 |
| WO2014071633A1 (en) | 2014-05-15 |
| EP2917335B1 (en) | 2018-04-04 |
| BR112015010698A2 (pt) | 2017-07-11 |
| AR093426A1 (es) | 2015-06-10 |
| EP2917335A1 (en) | 2015-09-16 |
| EP2917335A4 (en) | 2016-06-29 |
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