CN113939198A - 具有改善肝功能活性的菌株及其用途 - Google Patents
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Abstract
本发明涉及具有改善肝功能活性的菌株及其用途，具体地，本发明涉及用于改善肝功能的保健食品组合物及用于改善肝功能的药物组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。本发明的上述菌株抑制丙氨酸氨基转移酶(ALT，Alanine Aminotransferase)活性或天门冬氨酸氨基转移酶(AST，Aspartate Aminotransferase)活性、抑制肝内总胆固醇或中性脂肪的浓度增加、抑制肝内脂肪堆积的活性优秀，因此，可用于制备改善肝功能的食品组合物及药物组合物。
Description
技术领域
本申请要求2019年5月14日申请的韩国专利申请第10-2019-0056228号及2020年4月8日申请的韩国专利申请第10-2020-0042863号的优先权，以上说明书的全部内容为本申请的参考文献。
本发明涉及具有改善肝功能活性的菌株及其用途，具体地，本发明涉及利用保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株来改善肝功能的保健食品组合物及药物组合物。
背景技术
众所周知，肝脏与精神活动密切相关，在人体内起到储存并循环血液、调节血量、防御解毒的作用。随着工业化的发展，我们的身体始终暴露在污染物质、有毒物质中，因此，肝脏也在持续进行解毒作用。
并且，肝脏作为缓冲能力较大的器官，在疾病的初始阶段中难以表现，只有在明显恶化时才会被发现。肝硬化、肝癌等是各种肝脏疾病慢性进展的情况下共同导致的最终阶段。病因有酒精、药物、化学药品、病毒性肝炎、胆道疾病、血色病(hematochromatosis)等代谢性疾病及自身免疫性疾病，但在大多情况下病因不明，因此肝脏是早期健康管理上非常重要的器官。
根据最近调查的韩国人的健康水平，每10万人中因肝癌而死亡的有23.4人，位居世界第一，因慢性肝病死亡的有28.8人，位居世界第三。并且，最近韩国国家统计局宣布，肝病在韩国40岁人群中属于死亡率最高的死因，每10万人中有56.1例，因此，肝病成为重要的社会问题，最近因精神压力导致肝损伤的发病数量逐渐增加，正在进一步集中进行针对改善肝功能的研究。
另一方面，益生菌在我们的饮食生活中已经使用了很长时间，尤其，报道了益生菌具有肠道调理作用、抗癌效果及加强免疫效果等有益效果。
对此，本发明人在研究益生菌的过程中，尤其，利用在之前研究中分离并鉴定为新菌株的菌株，即，在针对保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株研究新生理活性的过程中，确认到上述细菌能够有效改善肝功能，从而完成了本发明。
现有技术文献
专利文献
专利文献1：韩国公开专利第10-2018-0103772号
发明内容
技术问题
因此，本发明的目的在于，提供用于改善肝功能的保健食品组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
本发明的另一目的在于，提供用于改善肝功能的药物组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
技术方案
为了实现上述本发明的目的，本发明提供用于改善肝功能的保健食品组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
根据本发明一实施例，上述组合物可抑制丙氨酸氨基转移酶(ALT，AlanineAminotransferase)活性或天门冬氨酸氨基转移酶(AST，Aspartate Aminotransferase)活性、抑制肝内总胆固醇或中性脂肪的浓度增加、或抑制肝内脂肪堆积。
根据本发明一实施例，上述保健食品组合物可以为选自粉末、颗粒、丸剂、片剂、胶囊、糖果、糖浆及饮料中的一种剂型。
并且，本发明提供用于改善肝功能的药物组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
根据本发明一实施例，上述组合物可抑制丙氨酸氨基转移酶(ALT，AlanineAminotransferase)活性或天门冬氨酸氨基转移酶(AST，Aspartate Aminotransferase)活性、抑制肝内总胆固醇或中性脂肪的浓度增加、或抑制肝内脂肪堆积。
并且，本发明提供肝功能改善方法，包括向个体施用或服用组合物的步骤，上述组合物包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
并且，本发明提供组合物的肝功能改善用途，上述组合物包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
发明的效果
本发明涉及用于改善肝功能的保健食品组合物及用于改善肝功能的药物组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分，本发明的上述菌株抑制丙氨酸氨基转移酶(ALT，Alanine Aminotransferase)活性或天门冬氨酸氨基转移酶(AST，AspartateAminotransferase)活性、抑制肝内总胆固醇或中性脂肪的浓度增加、抑制肝内脂肪堆积的活性优秀，因此，可用于制备改善肝功能的食品组合物及药物组合物。
附图说明
图1为本发明一实施例中利用保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株制备的用于改善肝功能的保健食品的形态的照片。
具体实施方式
本发明涉及具有肝功能改善活性的菌株的新用途，上述具有改善肝功能活性的菌株是指本发明的发明人通过之前首次分离并鉴定来公开的保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株。
本发明人为了确认本发明的上述菌株是否能够改善肝功能并具有保护肝的活性而进行了实验，结果，当向肝损伤的动物模型给药粉末化上述菌株制备的食品时，确认到保护肝细胞且抑制作为肝损伤指标的丙氨酸氨基转移酶活性及天门冬氨酸氨基转移酶活性的效果优秀。
上述丙氨酸氨基转移酶(ALT，Alanine Aminotransferase)是作为代表肝损伤的指标存在于肝脏、肾脏、心脏、肌肉等的酶，其中，存在于肝脏的量最多，当肝细胞受损时，由于通过应激反应导致其数值增加，因此用作肝病的指标。当肝脏主要因各种原因而受损时，丙氨酸氨基转移酶会被释放到血液，从而导致血液中数值上升。
并且，作为另一肝损伤指标的天门冬氨酸氨基转移酶(AST，AspartateAminotransferase)与上述丙氨酸氨基转移酶相同，当肝脏受损时，因释放到血液而导致血液中数值相对于正常状态有所上升，因此，可通过测定这种指标的含量来确认肝损伤程度。
另一方面，当向诱发肝损伤的小鼠组给药作为本发明菌株的保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株时，结果确认到血液中增加的丙氨酸氨基转移酶及天门冬氨酸氨基转移酶的数值减少2倍以上的效果。
并且，在本发明一实施例中，分析了当给药本发明的菌株时是否能够减少因脂肪肝发病导致肝内增加的总胆固醇和中性脂肪的含量，结果确认到当向脂肪肝动物模型给药本发明的菌株时可有效抑制肝内总胆固醇和中性脂肪的含量并抑制肝内脂肪堆积。
众所周知，脂肪肝是因肝脏中的脂蛋白形成及释放受到抑制或中性脂肪(triglyceride)合成增加导致的。乙硫氨酸(Ethionine)通过抑制RNA和蛋白质的合成来减少肝内腺苷三磷酸(ATP)的生成，从而能够抑制中性脂肪向血浆的释放并通过抑制脂蛋白的合成来再次抑制肝的脂质释放，因此，当给药乙硫氨酸时，随着肝组织内中性脂肪堆积，因血液中的中性脂肪及总胆固醇的含量急剧降低而引起微泡性脂肪病(microvasicularsteatosis)。由此，在通过乙硫氨酸诱发的脂肪肝模型中，将肝组织内中性脂肪含量及血清内总胆固醇含量用作模型的指标。
基于如上所述的内容，可确认到本发明的菌株具有减少肝组织内中性脂肪及总胆固醇的效果，因此，可用于脂肪肝的改善及治疗。
并且，在本发明的另一实施例中，根据临床试验审查委员会(IRB)的规定以志愿人员为对象进行临床试验的结果，相比于未摄取的组，摄取利用本发明的菌株制备的保健食品的组的血液中的丙氨酸氨基转移酶活性及天门冬氨酸氨基转移酶活性受到抑制，由此可知，本发明的菌株具有改善肝功能的效果，并且，通过摄取本发明的菌株还可降低总胆固醇及低密度脂蛋白胆固醇的含量。
由此，基于上述结果，本发明人已知可将本发明人通过之前首次分离并鉴定来公开的保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株用于改善肝功能的用途使用。
因此，本发明可提供用于改善肝功能的保健食品组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
本发明的上述用于改善肝功能的保健食品组合物中可能包含的上述菌株数量可以为1.0×105CFU/g～2.0×1015CFU/g，本发明的保健食品组合物可包含上述菌株中的一种菌株，或者可包含一种以上的混合菌株，最优选地，可包含混合上述三种菌株的混合菌株。
在本发明的实验中确定的上述保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株的肝功能改善效果，即，抑制血液中的丙氨酸氨基转移酶活性或天门冬氨酸氨基转移酶活性效果、抑制肝组织内总胆固醇及中性脂肪效果、抑制肝内脂肪堆积效果还可通过单独处理上述菌株来实现，当混合两种以上的上述菌株时，可获得更优秀的效果，当混合三种菌株时，可获得最优秀的效果。
本发明的上述食品组合物包含功能食品(functional food)、营养补充剂(nutritional supplement)、保健食品(health food)及食品添加剂(food additives)等所有形式。以上类型的食品组合物可通过所属领域中通常使用的公知方法制备成多种形式。例如，保健食品可以为利用本发明的菌株本体、其破碎物及上述菌株培养物组成的组中的一个以上制成的茶、果汁及饮用剂等形式的饮品，或者可制成颗粒、胶囊及粉末摄取。并且，可通过向本发明的菌株、其破碎物及培养物组成的组中的一个以上混合具有肝功能改善效果的公知物质或活性成分来将其制备成组合物形式。并且，功能食品可通过向饮料(包括酒精饮料)、水果及其加工食品(例如，水果罐头、瓶装罐头、果酱、橘子酱等)、鱼类、肉类及其加工食品(例如，火腿、玉米牛肉香肠等)、面包类及面类(例如，乌冬面、荞麦面、拉面、意大利面、通心粉等)、果汁、各种饮用剂、饼干、饴糖、乳制品(例如，黄油、奶酪等)、食用植物油、人造黄油、植物蛋白、软罐头、冷冻食品、各种调味料(例如，大酱、酱油、酱汁等)等添加本发明的菌株、其破碎物及培养物组中的一种以上来制备。
在本发明的食品组合物中，虽然针对本发明的上述菌株、其破碎物及培养物等的含量未做特别限制，但是，优选为最终制备食品的0.01重量百分比至50重量百分比。并且，本发明的菌株、其破碎物及其培养物可被制备成粉末或浓缩液形式，以便用作食品添加剂的有效成分。
并且，本发明可提供用于改善肝功能的药物组合物，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
上述药物组合物可单独包含药学上可接受的盐，或者，还可包含一种以上药学上可接受的载体、赋形剂或稀释剂。例如，药学上可接受的载体还可包括口服给药用载体或肠胃外给药用载体。作为口服给药用载体可包括乳糖、淀粉、纤维素衍生物、硬脂酸镁、硬脂酸等。并且，作为肠胃外给药用载体可包括水、合适的油、盐水、水溶性葡萄糖及甘醇等，还可包括稳定剂及保鲜剂。作为合适的稳定剂有亚硫酸氢钠、亚硫酸钠或抗坏血酸等抗氧化剂。作为合适的保鲜剂有苯扎氯铵、羟苯甲酯或羟苯丙酯及氯丁醇。除此之外的药学上可接受的载体可参照以下文献的记载(Remington's Pharmaceutical Sciences，19th ed.，MackPublishing Company，Easton，PA，1995)。
本发明的药物组合物可通过任何方法向包括人在内的哺乳动物给药。例如，可通过口服或肠胃外给药方式给药。作为肠胃外给药方法有静脉内、肌肉内、动脉内、骨髓内、硬膜内、心脏内、经皮、皮下、腹腔内、鼻腔内、肠道、局部、舌下或直肠内给药的方式，但并不限定于此。优选地，本发明的药物组合物可口服或经皮给药。
本发明的药物组合物可基于如上所述的给药途径制备成用于口服给药用制剂或肠胃外给药用制剂。
当制备口服给药用制剂时，本发明的组合物可通过所属领域的公知方法制剂成粉末、颗粒、片剂、丸剂、糖衣片剂、胶囊剂、液剂、胶剂、糖浆剂、泥浆剂、悬浮剂等。例如，作为口服用制剂可通过混合活性成分与固体赋形剂并粉碎其添加合适的助剂加工成颗粒混合物来制备片剂或糖衣片剂。例如，合适的赋形剂可以为包括乳糖、葡萄糖、蔗糖、山梨糖醇、甘露糖醇、木糖醇、赤藓糖醇及麦芽糖醇等的糖类；包括玉米淀粉、小麦淀粉、大米淀粉及马铃薯淀粉等的淀粉类；包括纤维素、甲基纤维素、羧甲基纤维素钠及羟丙基甲基纤维素等的纤维素类；以及包括明胶、聚乙烯吡咯烷酮等的填充剂。并且，可根据情况添加交联聚乙烯吡咯烷酮、琼脂、海藻酸或海藻酸钠等作为崩解剂。而且，本发明的药物组合物还可包含抗凝剂、润滑剂、润湿剂、调味剂、乳化剂及防腐剂等。
当制备肠胃外给药用制剂时，可通过所属领域的公知方法制剂成注射剂、霜剂、乳液剂、外用软膏剂、油剂、保湿剂、凝剂、气溶剂及鼻腔吸入剂的形式。上述剂型均记载于制药化学中作为通常使用的公知处方书的文献(Remington's Pharmaceutical Science，15th Edition，1975.Mack Publishing Company，Easton，Pennsylvania 18042，Chapter87：Blaug，Seymour)。
本发明的菌株本体、其破碎物及上述菌株培养物等的总有效量可通过单剂量(single dose)向患者给药，还可通过以多剂量(multiple dose)长时间给药的分次治疗方案(fractionated treatment protocol)给药。本发明的药物组合物可基于疾病的严重程度改变有效成分的含量。优选地，每日按1kg的患者体重，本发明的菌株、其破碎物及培养物的总用量约为0.01ug至1000mg，最优选地，可以为0.1ug至100mg。但由于，除药物组合物的给药途径及治疗次数外，本发明的上述菌株本体、其破碎物及上述菌株培养物等的用量应基于患者的年龄、体重、健康状态、性别、疾病严重程度、饮食及排泄率等多种因素来确定患者的有效给药量，在考虑以上因素的情况下，本发明所属领域的普通技术人员可通过将本发明的菌株、其破碎物及培养物作为免疫功能增强剂的特定用途来确定适当的有效给药量。因此，针对其剂型、给药途径及给药方法未做特别限制，只要本发明的药物组合物表现本发明的效果即可。
而且，在本发明中，肝功能改善是指治疗、减轻、预防肝损伤带来的症状，具有治疗、减轻或预防肝功能、肝状态、肝疾、肝病的含义。
在本发明中，因肝损伤引起的肝病还可包括自身免疫性肝病、药物性肝病、酒精性肝病、传染性肝病、遗传代谢性肝病、急性肝病、肝炎、胆汁淤积、脂肪肝及慢性肝病，但并不限定于此。
以下，通过实施例进一步详细说明本发明。这种实施例仅用于进一步具体说明本发明，这种实施例并不限定本发明的范围。
准备例
1.制备包含本发明菌株的试验用食品
本发明人委托菌株保藏机构向作为之前分离并鉴定的新菌株的三种菌株赋予了保藏编号，并利用所保藏的三种菌株(戊糖乳杆菌KF8、枯草芽孢杆菌KF11、乳酸乳球菌KF140)委托Mediogen公司制备试验用食品。
在本发明中所使用的三种菌株如下所示。
戊糖乳杆菌KF8：在2017年3月24日，由国际微生物保藏中心韩国微生物保藏中心(KCCM)所保藏，保藏编号为KCCM11997P的菌株。
枯草芽孢杆菌KF11：在2017年2月24日，由国际微生物保藏中心韩国微生物保藏中心(KCCM)所保藏，保藏编号为KCCM119981P的菌株。
乳酸乳球菌KF140：由国际微生物保藏中心韩国微生物保藏中心(KCCM)所保藏，保藏编号为KCCM11673P的菌株。
利用以上菌株制备的试验用食品通过现有的菌株产品制备方法委托Mediogen公司制备而成，主要原料包括麦芽糊精、无水结晶葡萄糖及本发明的三种菌株(分别2.0×109CFU/g)，以1.5g/袋为单位进行包装并冷冻保管直至用于试验。
在此情况下，在培养基中针对用于制备上述试验用食品的各个上述菌株进行培养并获取菌株后，通过冷冻干燥将其粉末化，从而使用粉末形式的菌株。
为了用在本发明而制备的上述试验用食品的照片如图1所示。
2.试验对象
为了确认本发明的三种菌株对于肝功能改善的效果，筛选如下所述的试验对象。
筛选符合以下标准且自愿接受人体适用试验招募的对象，并仅对通过筛选标准的对象进行如下试验。以居住在大邱庆北地区且年龄按周岁在25岁至45岁之间的44名成人为对象进行筛选检查，从而筛选出符合筛选标准且不符合排除标准的36名对象。
对象筛选标准仅限于在试验期间能够进行追踪观察的对象及在听到有关研究目的和内容的充分说明后自愿签署试验同意书的对象。
另一方面，排除符合如下事项的对象。
-当摄取菌株产品时存在过敏反应的对象
-在最近一个月以内摄取处方药物、普通药品、保健功能食品或益生菌产品的对象
-在参加试验一个月以内摄取抗生素的对象
-在参加试验一个月以内曾做过肠胃或大肠内窥镜检查的对象
-BMI小于20或大于35的对象
-高血压(收缩压为160mmHg或舒张压100mmHg以上)对象
-空腹血糖浓度高于110的对象
-丙氨酸氨基转移酶或天门冬氨酸氨基转移酶超过正常上限2倍的对象
-处于妊娠期或哺乳期的女性
-在试验期间未同意用医学证明方法(例如，避孕套、套环、女性避孕套等)进行避孕的受孕期女性
-因患有厌食症、抑郁症、躁郁症、精神疾病等而需要持续治疗的对象。
-免疫相关疾病、重症的肝肾功能衰竭、恶性肿瘤、肺部疾病、胶原病、多发性硬化症、过敏性皮肤病及其他自身免疫性疾病等全身性疾病患者
-有可能对试验食物的吸收产生影响的存在肠胃相关疾病史或肠胃相关手术史的(排除仅切除阑尾手术或疝气手术)对象
-在参加试验的三个月以内参照其他人体适用试验或临床试验而摄取过其他试验用产品的对象(排除化妆品人体适用试验)
-除以上事项外，研究人员判断难以进行试验的对象
并且，通过入院当天的问诊从本试验中排除符合以下条件的对象。
-在入院前的2周内一次性摄取50g以上酒精(烧酒1瓶以上或啤酒2瓶以上)的饮酒对象
-在入院前的1周内一天饮用大于1杯烧酒或1杯啤酒的饮酒对象
-在入院前的1周内摄取乳制品、汉堡、比萨等快餐食品、速食食品、油炸类、烧烤类等的对象
-在上一次访问日之后摄取抗生素的对象
-在上一次访问日之后进行肠胃或大肠内窥镜检查的对象
-在上一次访问日之后摄取处方药品、普通药品或保健功能食品的对象
以上述条件筛选的对象作为可分析有效性的最少对象数量，在每个实验组中各为10名。
可根据临床试验审查委员会(IRB)的规定针对已筛选对象进行临床试验。
实施例1
分析摄取本发明菌株对于肝功能的改善效果
以上述试验对象人员为对象，分别利用在摄取菌株26天前后所采集的血液来测定血液内的天门冬氨酸氨基转移酶、丙氨酸氨基转移酶、总胆固醇、中性脂肪(Triglyceride)、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇的变化。
表1
血液内成分分析结果
分析结果如上述表1所示，在分别摄取作为本发明菌株的戊糖乳杆菌KF8、枯草芽孢杆菌KF11及乳酸乳球菌KF140的组中，相比于摄取上述菌株前，作为肝功能指标的丙氨酸氨基转移酶(ALT，Alanine Aminotransferase)及天门冬氨酸氨基转移酶(AST，AspartateAminotransferase)的数值得到有效抑制。
根据这种结果，本发明人确认了可将戊糖乳杆菌KF8、枯草芽孢杆菌KF11及乳酸乳球菌KF140的菌株用于改善肝功能及肝疾病的预防及治疗。
实施例2
分析肝损伤诱发动物模型摄取本发明菌株对于肝功能的改善效果
作为实验动物使用体重200g的雄性SD(Sprague-Dawley)大鼠(SLC，Japan)，向各个实验组分配5只用于实验，各个实验组如下所示。
表2
| 给药内容 | |
| 正常组 | 正常饮食摄取 |
| 对照组 | 正常饮食+腹腔给药四氯化碳 |
| 实验组1 | 正常饮食+腹腔给药四氯化碳+给药戊糖乳杆菌KF8 |
| 实验组2 | 正常饮食+腹腔给药四氯化碳+给药枯草芽孢杆菌KF11 |
| 实验组3 | 正常饮食+腹腔给药四氯化碳+给药乳酸乳球菌KF140 |
在此情况下，将上述四氯化碳与玉米油混合的50％溶液以0.75ml/kg的用量每周2次向大鼠腹腔给药并持续4周，在上述实验组中，将按照准备例制备的包含本发明菌株的功能食品粉末以每50mg/kg悬浮在1％CMC溶液，并将其口服给药每日一次，一周六次。随后，持续给药4周后，致死动物并获取血清，为了测定本发明菌株对于因四氯化碳引起的肝损伤的治疗效果而测定丙氨酸氨基转移酶活性及天门冬氨酸氨基转移酶活性。
表3
丙氨酸氨基转移酶及天门冬氨酸氨基转移酶浓度测定结果
分析结果如上述表3所示，在摄取本发明菌株的小鼠组中，因四氯化碳上升的血清内的丙氨酸氨基转移酶及天门冬氨酸氨基转移酶得到有效抑制。
这种结果说明本发明的戊糖乳杆菌KF8、枯草芽孢杆菌KF11及乳酸乳球菌KF140菌株对于肝毒性的肝保护效果及肝功能效果优秀。
实施例3
分析脂肪肝动物模型摄取本发明菌株对于肝功能的改善效果
作为实验动物使用体重20g的雄性C57BL/6小鼠(中型，韩国)，向各个实验组分配9只用于实验，各个实验组如下所示。作为脂肪肝诱发物质以颗粒形式提供高脂肪饮食(40％fat)，以10mg/kg的用量每日口服给药CML反应物。随后，向每只小鼠给药本发明菌株并分析脂肪肝改善程度。
表4
| 给药内容 | |
| 正常组 | 正常饮食摄取 |
| 对照组 | 高脂肪饮食+CML饮食 |
| 实验组1 | 高脂肪饮食+CML饮食+给药戊糖乳杆菌KF8 |
| 实验组2 | 高脂肪饮食+CML饮食+给药枯草芽孢杆菌KF11 |
| 实验组3 | 高脂肪饮食+CML饮食+给药乳酸乳球菌KF140 |
在此情况下，将每1×107CFU的本发明菌株悬浮于0.9％的盐水溶液，并将其口服给药每日一次，一周六次。随后，持续给药4周后，致死动物并采取血液，测定肝功能相关酶和中性脂肪(Triglyceride)的含量。
表5
肝功能相关酶及中性脂肪浓度测定结果
分析结果如上述表7所示，在摄取本发明菌株的小鼠组中，因肝损伤而增加的天门冬氨酸氨基转移酶和丙氨酸氨基转移酶的数值得到有效抑制，并且，确认到防止增加中性脂肪含量的效果。
根据这种结果，本发明人确认了本发明的戊糖乳杆菌KF8、枯草芽孢杆菌KF11或乳酸乳球菌KF140菌株可通过预防因摄取糖化毒素产生的肝损伤来有效改善并治疗脂肪肝等肝疾病。
以上，以优选实施例为中心说明了本发明。应当理解的是，本发明所属领域的普通技术人员可在不脱离本发明的基本特征的范围内通过变形的实施方式实现本发明。因此，应基于所说明的观点考虑在此公开的实施例，而并非限制性观点。本发明的范围应基于发明要求保护范围表示，而并非上述说明，与其等同范围内的所有不同应以均属于本发明的含义加以解释。
Claims (7)
1.一种用于改善肝功能的保健食品组合物，其特征在于，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
2.根据权利要求1所述的用于改善肝功能的保健食品组合物，其特征在于，上述组合物抑制丙氨酸氨基转移酶活性或天门冬氨酸氨基转移酶活性、抑制肝内总胆固醇或中性脂肪的浓度增加、或抑制肝内脂肪堆积。
3.根据权利要求1所述的用于改善肝功能的保健食品组合物，其特征在于，上述保健食品组合物为选自粉末、颗粒、丸剂、片剂、胶囊、糖果、糖浆及饮料中的一种剂型。
4.一种用于改善肝功能的药物组合物，其特征在于，包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
5.根据权利要求4所述的用于改善肝功能的药物组合物，其特征在于，上述组合物抑制丙氨酸氨基转移酶活性或天门冬氨酸氨基转移酶活性、抑制肝内总胆固醇或中性脂肪的浓度增加、或抑制肝内脂肪堆积。
6.一种肝功能改善方法，其特征在于，包括向个体施用或服用组合物的步骤，上述组合物包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
7.一种组合物的肝功能改善用途，其特征在于，上述组合物包含选自由保藏编号为KCCM11997P的戊糖乳杆菌KF8菌株、保藏编号为KCCM11981P的枯草芽孢杆菌KF11菌株及保藏编号为KCCM11673P的乳酸乳球菌KF140菌株组成的组中的一种以上菌株、其破碎物或其培养物作为有效成分。
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| CN117100774B (zh) * | 2023-10-25 | 2024-02-13 | 山东中科嘉亿生物工程有限公司 | 嗜酸乳杆菌jyla-16在制备治疗胆结石的产品中的应用 |
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| EP3970511A1 (en) | 2022-03-23 |
| US20220211781A1 (en) | 2022-07-07 |
| JP2022532643A (ja) | 2022-07-15 |
| EP3970511A4 (en) | 2023-01-11 |
| WO2020231127A1 (ko) | 2020-11-19 |
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