CN116887848A - 对抗肠屏障功能障碍和热应激的益生菌组合物和方法 - Google Patents
对抗肠屏障功能障碍和热应激的益生菌组合物和方法 Download PDFInfo
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Abstract
本发明涉及一种包含两种或更多种细菌菌株或由其组成的组合物，所述组合物被配制用于预防或治疗受试者的肠屏障功能障碍和/或热应激。更具体地，所述组合物包含罗伊氏乳杆菌MM2‑3(Lactobacillus reuteri MM2‑3)、植物乳杆菌WCSF1(Lactobacillus plantarum WCSF1)、嗜热链球菌B/R(Streptococcus thermophilus B of R)和大肠杆菌Nissle1917(Escherichia coli Nissle 1917)中的两种或更多种。本发明还包括本发明治疗诸如炎性肠病(IBD)和热应激的病症的用途。
Description
技术领域
本发明涉及一种包含两种或更多种细菌菌株或由其组成的组合物，所述组合物被配制用于预防或治疗受试者的肠屏障功能障碍和/或热应激。更具体地，所述组合物包含罗伊氏乳杆菌MM2-3(Lactobacillus reuteri MM2-3)、植物乳杆菌WCSF1(Lactobacillusplantarum WCSF1)、嗜热链球菌B/R(Streptococcus thermophilus B of R)和大肠杆菌Nissle1917(Escherichia coli Nissle 1917)中的两种或更多种。本发明还包括本发明治疗诸如炎性肠病(IBD)和热应激的病症的用途。
背景技术
益生菌是当以足够的量施用时赋予宿主有益作用的活微生物。长时间的热应激会引起一种称为“漏肠”的症状，其中增加的来自肠腔的过多分子进入血流引起各种健康问题。在极端情况下，热应激也会引起死亡。
长时间的体力消耗会使整个身体发生若干严重变化，所述变化是由急性期蛋白表达增加和激素释放变化引起的，从而导致肌肉损伤、肠通透性增加以及甚至全身性炎症[Clark,A.和Mach,N.,J Int Soc Sports Nutr 13:43(2016)]。具体而言，经受户外训练的个体诸如士兵和运动员，尤其是在炎热和潮湿的条件下，通常具有升高的热应激损伤风险。热应激可能削弱肠道上皮细胞并且允许肠道中的细菌毒素进入原本无菌的血流中，从而引起严重的中暑相关症状并且甚至死亡[Moran,A.P.,Prendergast,M.M.和Appelmelk,B.J.FEMS Immunol Med Microbiol 16:105-115(1996)]。
已知益生菌用于多种应用[Cheng,F.S.等人,World J Clin Cases 8:1361-1384(2020)；Cinque,B.等人,PLoS One 11:e0163216,doi:10.1371/journal.pone.0163216(2016)；Kumar,M.等人,Am J Physiol Gastrointest Liver Physiol 312:G34-G45,doi:10.1152/ajpgi.00298.2016(2017)；Shing,C.M.等人,Eur J Appl Physiol 114:93-103,doi:10.1007/s00421-013-2748-y(2014)]。
需要一种改善的益生菌，所述益生菌预防、改善或治疗肠屏障功能障碍和/或由热应激引起的活动能力相关问题。
发明内容
本发明涉及一种新型益生菌混合剂，其改善患有肠屏障功能障碍和/或由热应激引起的活动能力相关问题的个体的健康状况。在这项研究中，本发明诸位发明人配制了一种益生菌混合剂，其改善肠屏障功能和热应激下的身体活动能力。首先，本发明诸位发明人评价了一组益生菌菌株并且验证了强化肠上皮内层的益生菌菌株。接下来，本发明诸位发明人使用四种表现最佳的菌株罗伊氏乳杆菌MM2-3(Lactobacillus reuteri MM2-3)、植物乳杆菌WCSF1(Lactobacillus plantarum WCSF1)、嗜热链球菌B/R(Streptococcusthermophilus B of R)和大肠杆菌Nissle 1917(Escherichia coli Nissle 1917)配制了益生菌混合剂。在动物模型中，4菌株益生菌混合剂降低了肠通透性，促进了紧密连接蛋白的表达，并且改善了身体活动能力，表明了对抗热应激的保护功效。这种保护功效比单独菌株的保护功效高多达5.5倍。我们的益生菌混合剂适用于解决由肠屏障功能障碍和热应激引起的健康问题以及促进热应激下的身体活动能力。
在第一方面，本发明提供了一种组合物，所述组合物包含罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌B/R和大肠杆菌Nissle 1917中的两种或更多种或由其组成，所述组合物被配制用于预防或治疗受试者的肠屏障功能障碍和/或热应激。
在一些实施方案中，所述组合物包含罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌B/R和大肠杆菌Nissle 1917或由其组成。
在一些实施方案中，所述热应激至少部分是由于所述受试者的体力消耗。
在一些实施方案中，所述组合物被配制用于预防或治疗所述受试者的肠屏障功能障碍和/或热应激诱发的肠道通透性。
在一些实施方案中，所述组合物包含约108菌落形成单位(CFU)的剂量的每种菌株。
在一些实施方案中，所述受试者是人。
在一些实施方案中，所述组合物呈明胶胶囊、压制片剂、胶囊锭、液体饮料或小药囊的形式。
在第二方面，本发明提供了一种治疗或预防方法，所述方法包括向需要这种治疗或预防的受试者施用有效量的第一方面的组合物。
在一些实施方案中，所述受试者患有或将患有肠屏障功能障碍和/或热应激。
在一些实施方案中，向所述受试者每天施用包含至少108至1011CFU(菌落形成单位)的组合物。
在一些实施方案中，向所述受试者施用包含至少108CFU(菌落形成单位)的每种菌株、优选至少5x109CFU(菌落形成单位)的每种菌株的组合物。
在一些实施方案中，所述组合物降低核心体温并且提高跑步活动能力。如本文所示，包含罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌B/R和大肠杆菌Nissle 1917的组合物与未处理相比使增加了约1.5倍的跑步时间。所述组合物还使锻炼期间的核心温度降低约1℃。
在一些实施方案中，所述组合物降低肠上皮层的通透性。如本文所示，与未处理相比，包含罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌B/R和大肠杆菌Nissle 1917降低了炎性肠病模型中炎症下的上皮层通透性。
在一些实施方案中，所述受试者是人。
在第三方面，本发明提供了在第一方面中定义的组合物，所述组合物用于治疗或改善受试者的与肠屏障功能障碍和/或热应激相关的病症的方法中。
在第四方面，本发明提供了第一方面中定义的组合物用于制造用于治疗或预防受试者的肠屏障功能障碍和/或热应激的药剂的用途。
在一些实施方案中，所述治疗针对热应激诱发的肠道通透性。
在一些实施方案中，所述药剂降低受试者的核心体温并且提高跑步活动能力。
在一些实施方案中，所述药剂是用于人的。
现在已经大体上描述了本发明，通过参考以下实施例将更容易地理解本发明，所述实施例是通过说明的方式来提供的，并不旨在限制本发明。
附图说明
图1示出了在与单一益生菌菌株(A)和益生菌混合剂(B)共培养的Caco-2细胞中对抗热应激的紧密连接蛋白ZO-1的表达的分析。进行蛋白质印迹以分析ZO-1表达(变化倍数)，相对于看家蛋白β-肌动蛋白归一化。将ZO-1表达设置为1。将107CFU/mL的4菌株混合剂(包含罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌和大肠杆菌Nissle1917Eda，CFU/mL相等)用于测定。
图2示出了动物模型的核心体温(A)和跑步时间长度(B)。将每组中的单独动物用“X”表示，其中对照组在左并且处理组在右。与未处理的动物(蓝色)相比，用益生菌混合剂处理的动物(右)展现出更低的平均核心温度并且锻炼了更长的时间段(p<0.05)。
图3示出了经受跑步锻炼的给予益生菌混合剂的动物(橙色)和未处理动物(蓝色)的血清中FITC-葡聚糖浓度的测量。将每组中的单独动物用“X”表示。与给予益生菌混合剂的动物相比，在锻炼后未处理动物的血清中观察到FITC-葡聚糖的浓度显著增加，表明阴性对照动物的肠道通透性增加(p<0.01)。
图4示出了在经受跑步锻炼后，来自PBS处理的动物和用益生菌混合剂处理的动物的结直肠组织的免疫荧光染色。将细胞核(蓝色)用DAPI染色，而紧密连接蛋白闭合蛋白(用箭头指示)和ZO-1(红色)用它们各自的与荧光探针缀合的抗体检测。在荧光显微镜下观察荧光。
图5示出了通过H&E染色对结肠组织中炎症的组织学评估。在第11天收集结肠组织，拉紧并且根据显微术观察。对照，没有DSS。
图6示出了与幼稚CD4+T细胞共培养的发炎Caco-2上皮层的通透性的图。在用本发明益生菌或商业益生菌处理后，测量从顶端室穿过到基底室的FD10的浓度。显著性(P<0.05，T-检验)用星号指示。误差棒代表两个独立实验的标准偏差。
定义
为了方便起见，这里收集了说明书、实施例和所附权利要求中采用的某些术语。
如本说明书和所附权利要求中所用，单数形式的“一个/种(a)”、“一个/种(an)”和“所述”包括复数指示物，除非上下文清楚地另外规定。
如本文所用，术语“包含”或“包括”应解释为具体说明所提及的所陈述特征、整数、步骤或组分的存在，但是不排除一个或多个特征、整数、步骤或组分或其组的存在或添加。然而，在本公开文本的上下文中，术语“包含”或“包括”也包括“由……组成”。词语“包含(comprising)”的变化形式，例如“包含(comprise)”和“包含(comprises)”以及“包括(including)”的变化形式，例如“包括(include)”和“包括(includes)”具有相应变化的含义。
如本文所用的细菌“菌株”是指在生长或繁殖时保持遗传不变的细菌。
为了方便起见，在本说明书中提到的书目参考文献以参考文献列表的形式列出并且添加在实施例的末尾。此类文后参考书目的全部内容通过引用并入本文，但是在说明书中提及所述参考书目并非意指它们构成公知常识的一部分。
具体实施方式
实施例
大体上遵循本领域已知的并且未具体描述的标准分子生物学技术，诸如Green和Sambrook,Molecular Cloning:A Laboratory Manual,Cold Springs HarborLaboratory,纽约(2012)中所述。
实施例1
材料和方法
益生菌菌株
将一组益生菌菌株(表1)培养大约16小时，然后与人结直肠细胞(Caco-2)共培养或然后口服施用于小鼠。将益生菌培养物在无菌1x PBS中简单洗涤并且离心(4000rpm，4分钟)三次，然后使用分光光度计取得OD600读数。然后计算每种菌株的菌落形成单位(CFU)，并且将细胞在无菌1x PBS中稀释至所希望的CFU。对于体外共培养研究，每种菌株使用104、105和106CFU/mL。对于小鼠的口服施用，将罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌和大肠杆菌Nissle 1917以每种菌株5x108CFU/mL混合以获得益生菌混合剂。
表1.用于体外筛选的益生菌菌株和培养条件的列表
Caco-2和益生菌菌株的共培养
将Caco-2维持在加湿环境(37℃，5％CO2，95％空气)中的补充有15％FBS和0.1％v/v抗生素(青霉素/链霉素)的DMEM(Life Technologies)中。在补充有不含抗生素的DMEM的15％FBS(BioWest)中进行与益生菌菌株的共培养，并且在加湿环境(41℃，5％CO2，95％空气)中培养24小时，然后收获Caco-2细胞用于蛋白质提取。
蛋白质印迹分析
Caco-2细胞和相应益生菌菌株共培养24小时后，除去细胞培养物上清液，并且将Caco-2细胞用1x PBS轻轻洗涤。向每个样品中添加一百微升冰冷的1％Triton-X细胞裂解缓冲液(150mM NaCl，1％Triton-X，50mM Tris pH 8.0)，并且将细胞在冰上裂解。将裂解物在4℃下以12000rpm离心15分钟。收集上清液并且煮沸，用于在10％SDS-PAGE凝胶上分析。使用抗闭合小带1(anti-zona occludens 1)(抗ZO1)抗体(Life Technologies)检测从SDS-page凝胶转移到0.4μm PVDF膜上的蛋白质条带。
临床前动物实验
使BALB/c小鼠(雄性，8-9周，21-25克，活体)适应环境至少三天，然后皮下植入(SC)实时遥测温度记录器“Anipill”(Data Sciences International)。为了比较实际的核心温度，使五只动物经受记录器的腹膜内植入(IP)。将记录的SC和IP温度的平均值的差异添加到所有随后在动物中获得的SC温度读数中。术后允许动物恢复至少14天。然后，使它们经受连续五天的跑步机训练锻炼，所述跑步机训练锻炼改编自先前发表的小鼠跑步机疲劳测试[Dougherty,J.P.等人Journal of Visualized Experiments:JoVE,doi:10.3791/54052(2016)]。然后允许动物休息另外三天，然后在设定为32℃、相对湿度(RH)为60％-80％的环境室中进行实际的跑步机跑步实验。
在跑步实验前大约18小时，向一组动物口服管饲无菌PBS，而向另一组动物以109CFU/只动物给予益生菌混合剂。然后，将动物在跑步实验前禁食3小时，并且在跑步实验前1小时口服管饲4kDa FITC-葡聚糖(Sigma-Aldrich)。对于跑步实验，当记录的体温达到41℃时，或当动物在跑步机末端休息至少5秒钟时，或当动物跑了40分钟时(以先发生的为准)，将动物从跑步机上移除。跑步实验后，立即处死所有动物。取出躯干血和大肠用于随后的血清和组织学染色分析。
我们在第8-15天将小鼠用2％葡聚糖硫酸钠(DSS)处理。收集结肠组织，通过苏木精和伊红(H&E)染色进行炎症的组织学评估。所有涉及动物的程序均经IACUC批准(参考号R19-0435和2020/SHS/1614)。
血清分析
从每只动物取出血液样品后，立即将冰冷的抗凝剂(30％v/v)添加到所有血液样品中。通过将管翻转几次来轻轻混合血液样品，然后以2000rpm和4℃离心30分钟。然后小心地除去血清，并且使用酶标仪在激发475nm/发射515nm(BioTek)下将10μl血清用于FITC-葡聚糖分析。
组织学和共聚焦显微术
取出每只动物的大肠，并且用无菌1x PBS洗涤数次，直至干净。然后将每个样品小心盘绕并且置于冰冷的在蔗糖中的4％多聚甲醛(PFA)中，并且在旋转器上在4℃下固定过夜。第二天，然后将样品在30％蔗糖中洗涤至少12小时。小心地取出洗涤过的切片并且包埋在最佳切割温度化合物(OCT)中，然后在-80℃下储存，直到准备好用于通过低温恒温器的切片。将冷冻肠样品以7μm的厚度切片，并且封固在干净的显微镜载玻片上。允许切片周围过量的OCT在室温下解冻并且蒸发。将样品用0.1％BSA封闭30分钟，然后用抗闭合蛋白(Life Technologies)和抗ZO1抗体在4℃下孵育过夜。第二天，然后将样品在室温下与荧光探针缀合的二抗(Cell Signaling)一起孵育1小时，并且最后用细胞核染色DAPI(NucBlueTMFixed Cell ReadyProbesTMReagent,LifeTech)染色3分钟，然后将干净的盖玻片封固在每个样品上。在每次孵育之间以及在细胞核染色之前和之后，将样品用0.1％TBST轻轻洗涤。使用Prolong Gold防褪色封固剂(LifeTech)保存每个样品中的荧光。
实施例2
当与益生菌菌株共培养时，紧密连接蛋白在Caco-2中的表达增加
将生长至融合状态的Caco-2细胞与上述益生菌菌株在41℃下共培养24小时，以模拟热应激条件。进行蛋白质印迹以检测在有或没有益生菌的细胞培养物中紧密连接蛋白ZO-1的差异化表达。ZO-1是一种支架蛋白，其参与将上皮细胞保持在一起的各种紧密连接的形成[Fanning,A.S.等人,J Biol Chem 273:29745-29753(1998)]。几项研究显示，ZO-1蛋白的表达是温度和时间依赖性的[Dokladny,K.等人,Am J Physiol GastrointestLiver Physiol 290:g204-212(2006)；He,S.等人PLoS One 11:e0145236(2016)]。图1A示出了四种益生菌菌株罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌和大肠杆菌Nissle1917Eda在热应激条件下(41℃，24小时)在两种或三种不同浓度下分别使ZO-1表达增加1.3-4.8倍。此外，图1B示出了包含所有罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌和大肠杆菌Nissle 1917Eda 4的4-菌株混合剂使ZO-1表达增加超过7倍，比用单独菌株观察到的增加高1.5-5.5倍。
实施例3
益生菌混合剂改善动物模型中热应激诱发的肠道通透性
接下来，我们在动物模型中验证了4菌株混合剂的保护作用。为此，我们将四种益生菌菌株罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌和大肠杆菌Nissle 1917以5x108CFU/种菌株混合并且将混合剂施用于在炎热且潮湿的环境(32℃，RH 60％-80％)中经受跑步锻炼的小鼠。这项研究中使用的针对耐力的主要标志是核心体温和跑步时间长度。图2示出了用益生菌混合剂处理的动物具有低约1℃的核心温度和长1.5倍的跑步时间。我们还发现，益生菌混合剂处理的小鼠比PBS处理的对照多花6min达到峰值核心温度。这些结果揭示了与PBS处理的对照相比，我们的益生菌混合剂的处理提高了小鼠对抗热应激的身体活动能力。
我们还检查了4菌株益生菌混合剂对肠道上皮通透性的影响。当肠道上皮内层受损时出现“漏肠”综合征，允许增加分子和毒素外排而从肠道进入血流。核心体温的增加可能损害肠道上皮的完整性，从而引起“肠漏”综合征[Yang,P.C.等人,J Gasl Hepatol 22:1823-18311111(2007)]。我们在锻炼前一小时通过口服管饲施用荧光探针小分子FITC-葡聚糖，并且测量其在跑步实验后收集的血清中的浓度。如果肠道上皮受损，则可以检测到血清中FITC-葡聚糖浓度的增加。图3示出了用益生菌混合剂处理的动物的血清中的FITC-葡聚糖浓度比PBS处理的动物低2倍。此结果表明给予益生菌混合剂的动物的肠道通透性降低。此外，图4示出了在用益生菌混合剂处理的动物的肠道粘膜上皮中，紧密连接蛋白闭合蛋白和ZO-1的表达增加，表明与PBS处理的动物相比，肠道紧密连接在对抗热应激方面有所改善。
为了允许进一步验证我们的益生菌混合剂的功能，我们建立了炎性肠病(IBD)的体内模型。我们观察到肠长度减少以及DSS处理的小鼠的结肠组织中的溃疡和腺窝细胞损失，表明DSS处理成功诱导了炎症(图5)。
实施例4
在体外炎性肠病(IBD)模型中，益生菌混合剂降低了炎症上皮层的通透性
在体外炎性肠病(IBD)模型中测试本发明益生菌组合物治疗肠屏障功能障碍疾病诸如IBD的能力。
简言之，将5x104/cm2的Caco-2细胞接种在transwell上，并且诱导极化大约2周。在极化约12天后测量跨上皮电阻(TEER)，并且当电阻在800-1000Ohms/cm2之间时，极化的细胞准备用于随后的测定。将幼稚CD4+T细胞解冻并且维持在补充hIL-2(600U/mL)的ImmunoCultTMXF培养基(StemCell Technologies)中大约3天，然后与极化的Caco-2细胞共培养。用添加到顶端室(Caco-2)中的发炎混合剂(白介素1-β(rhIL-1β，25ng/ml)、肿瘤坏死因子-α(rhTNF-α，50ng/ml)、干扰素γ(rhIFN-γ，50ng/ml)和脂多糖(LPS，1μg/ml))使Caco-2/T细胞共培养物发炎，同时在基底室中培养T细胞。允许发炎36h，然后向经发炎的transwell补充相应的处理。
使用107CFU/mL的益生菌进行10h处理。从顶端室和基底室两者除去上清液，并且在新鲜细胞培养基中稀释1μg/mL的10kDa FITC-葡聚糖(FD10)，并且在每次指示处理结束时应用于顶端室。将无菌1X PBS补充到基底室中。将培养物在37℃下在5％CO2加湿气氛中孵育1h，然后从顶端室和基底室两者中除去100μL上清液。使用Synergy H1酶标仪(BioTek)在485nm的激发波长和525nm的发射波长下确定荧光强度。
结果显示，与未处理的对照相比，我们的益生菌混合剂的处理显著地导致基底室中的FD10低8.1％，表明通透性降低(图6)。使用可商购益生菌VSL#3(也称为Vivomixx)进行的对比处理导致基底室中的FD10高4.5％。总而言之，我们的结果显示，在体外IBD模型中，我们的益生菌混合剂发挥出对炎症下的上皮层的保护作用，这比VSL#3有利。
总结
我们验证了增加紧密连接蛋白ZO-1的表达的益生菌菌株。使用经验证的菌株，我们开发了一种新型的4菌株益生菌混合剂，并且在动物模型中证实了所述混合剂对抗肠屏障功能障碍和热应激的保护作用。所述4菌株混合剂(罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌B/R和大肠杆菌Nissle 1917)是新颖的并且能够促进紧密连接蛋白的表达，降低肠道通透性并且在炎热且潮湿的环境中提高身体活动能力(较低的核心体温和较长的跑步时间)。我们的益生菌混合剂适用于解决由肠屏障功能障碍和热应激引起的健康问题以及促进热应激下的身体活动能力。
参考文献
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Claims (18)
1.一种组合物，所述组合物包含罗伊氏乳杆菌MM2-3(Lactobacillus reuteri MM2-3)、植物乳杆菌WCSF1(Lactobacillus plantarum WCSF1)、嗜热链球菌B/R(Streptococcusthermophilus B of R)和大肠杆菌Nissle 1917(Escherichia coliNissle 1917)中的两种或更多种或由其组成，所述组合物被配制用于预防或治疗受试者的肠屏障功能障碍和/或热应激。
2.根据权利要求1所述的组合物，所述组合物包含罗伊氏乳杆菌MM2-3、植物乳杆菌WCSF1、嗜热链球菌B/R和大肠杆菌Nissle 1917或由其组成。
3.根据权利要求1或2所述的组合物，其中所述热应激至少部分是由于所述受试者的体力消耗。
4.根据权利要求1至3中任一项所述的组合物，所述组合物被配制用于预防或治疗所述受试者的热应激诱发的肠道通透性。
5.根据权利要求4所述的组合物，所述组合物包含约108菌落形成单位(CFU)的剂量的每种菌株。
6.根据权利要求1至5中任一项所述的组合物，其中所述受试者是人。
7.根据权利要求1至7中任一项所述的组合物，其中所述组合物呈明胶胶囊、压制片剂、胶囊锭、液体饮料或小药囊的形式。
8.一种治疗或预防方法，所述方法包括向需要这种治疗或预防的受试者施用有效量的根据权利要求1至7中任一项所述的组合物。
9.根据权利要求8所述的方法，其中所述受试者患有或将患有肠屏障功能障碍和/或热应激。
10.根据权利要求8或9所述的方法，其中向所述受试者每天施用包含至少108至1011CFU的组合物。
11.根据权利要求10所述的方法，其中向所述受试者施用包含至少108CFU的每种菌株、优选至少5x 109CFU的每种菌株的组合物。
12.根据权利要求8至11中任一项所述的方法，其中所述组合物改善肠屏障功能障碍和/或降低核心体温并且提高跑步活动能力。
13.根据权利要求8至12中任一项所述的方法，其中所述受试者是人。
14.根据权利要求1至7中任一项所述的组合物，所述组合物用于治疗或改善受试者的与肠屏障功能障碍和/或热应激相关的病症的方法中。
15.根据权利要求1至7中任一项所述的组合物用于制造用于治疗或预防受试者的肠屏障功能障碍或热应激的药剂的用途。
16.根据权利要求15所述的用途，其中所述治疗针对热应激诱发的肠道通透性。
17.根据权利要求15或16所述的用途，其中所述药剂改善所述受试者的肠屏障功能障碍或降低核心体温并且提高跑步活动能力。
18.根据权利要求15至17中任一项所述的用途，其中所述药剂是用于人的。
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