CN116236510A - 一种改善慢性酒精诱导的肝疾病的益生菌剂及其应用 - Google Patents
一种改善慢性酒精诱导的肝疾病的益生菌剂及其应用 Download PDFInfo
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Abstract
本发明涉及一种改善慢性酒精诱导的肝疾病的益生菌剂及其应用,所述益生菌剂中的菌株包括保藏编号为CGMCC No.1.12735的嗜酸乳杆菌Lactobacillus acidophilus LA85菌株和保藏编号为CGMCC No.10452的长双歧杆菌Bifidobacterium longum BL21菌株。两种菌株能够相互配合、相互促进,在改善慢性酒精诱导的肝疾病的效果上协同增效,相较于单菌使用,两种的组合方式在上述功效上更加显著,为改善慢性酒精诱导的肝疾病提供了新的策略,可以用于制备预防、缓解或治疗慢性酒精诱导的肝疾病的相关产品。
Description
技术领域
本发明属于益生菌剂技术领域,涉及一种改善慢性酒精诱导的肝疾病的益生菌剂及其应用。
背景技术
过量饮酒会导致脂肪肝(肝脂肪变性)的发生与发展。尽管轻度肝脏脂肪病变最初被认为一般是良性的,但越来越多的证据表明,脂肪积累的增加会对肝脏造成二次打击,导致发展为更严重的肝脏疾病,如脂肪性肝炎、纤维化、肝硬化甚至肝癌。
消除或抑制肝脏脂肪积累可能是抑制/治疗酒精性肝病ALD的一种有潜在价值的策略。已有研究表明,给予益生菌可以改善酒精性脂肪性肝炎患者的肝酶,并逆转酒精诱导的脂肪肝和肝损伤。酒精诱导的脂肪堆积是由于原位脂肪生成的增加和脂肪酸β-氧化的减少。
AMP活化蛋白激酶(AMPK)是一种关键的代谢主开关,可磷酸化许多组织中涉及脂质代谢的靶酶,包括肝脏。它通过灭活乙酰辅酶a羧化酶(ACC)来增加脂肪酸氧化。ACC是肝脏脂肪酸生物合成的限速酶,它催化乙酰辅酶a转化为丙二酰辅酶a,丙二酰辅酶a是脂肪酸合成的前体。此外,AMPK还调节固醇调节元件结合蛋白-1(SREBP-1)和过氧化物酶体增殖物激活受体-α(PPAR-α),这两种转录因子在肝脏和其他组织中负责胆固醇、脂肪酸和甘油三酯合成的酶的调节中起着关键作用。酒精摄入会导致AMPK活性降低,并增加肝脏中的脂肪积累。
相关机制研究表示,益生菌,尤其是双歧杆菌、乳杆菌的使用可以减弱酒精诱导的肠屏障功能障碍和内毒素血症(可能引发肝脏炎症反应等),另外,益生菌预处理可增加肝脏AMPK磷酸化和PPAR-α表达,降低SREBP-1表达,从而减轻酒精诱导的肝脏脂肪变性和损伤。双歧杆菌和乳杆菌在一些功能特性上具有特异性,特异性产生一些有益成分,功能因子等,若能联合使用双歧杆菌和乳杆菌,协同增效,共同作用发挥各自的有益作用,或许可以提高益生菌在慢性酒精诱导的肝疾病中应用前景。
发明内容
针对现有技术的不足,本发明的目的在于提供一种改善慢性酒精诱导的肝疾病的益生菌剂及其应用。
为达到此发明目的,本发明采用以下技术方案:
第一方面,本发明提供一种改善慢性酒精诱导的肝疾病的益生菌剂,所述改善慢性酒精诱导的肝疾病的益生菌剂中的菌株包括保藏编号为CGMCC No.1.12735的嗜酸乳杆菌Lactobacillus acidophilus LA85菌株和保藏编号为CGMCC No.10452的长双歧杆菌Bifidobacterium longum BL21菌株。
本发明创造性地将嗜酸乳杆菌LA85菌株和长双歧杆菌BL21菌株进行复配,发现两者能够相互配合、相互促进,在改善慢性酒精诱导的肝疾病的效果上协同增效,具体表现在:能够在减少慢性酒精诱导的肝脂积累和肝损伤上协同增效;能够在调节慢性酒精诱导的肝疾病小鼠的肝脂质代谢水平上协同增效;能够在减轻慢性酒精诱导的肝疾病小鼠肝细胞凋亡程度上协同增效。相较于单菌使用,两种的组合方式在上述功效上更加显著,获得了本领域技术人员难以预见的技术效果,为改善慢性酒精诱导的肝疾病提供了新的策略,可以用于制备预防、缓解或治疗慢性酒精诱导的肝疾病的相关产品,具有广泛的应用前景。
另外,嗜酸乳杆菌和长双歧杆菌均是益生菌,因此其在用于制备预防、缓解或治疗慢性酒精诱导的肝疾病的相关产品时,安全性高,且不易产生抗药性。
优选地,在益生菌剂中,所述嗜酸乳杆菌Lactobacillus acidophilus LA85菌株的活菌数不低于1×109CFU/mL,例如1×109CFU/mL、3×109CFU/mL、5×109CFU/mL、8×109CFU/mL、1×1010CFU/mL、5×1010CFU/mL、1×1011CFU/mL、1×1012CFU/mL等;所述长双歧杆菌Bifidobacterium longum BL21菌株的活菌数不低于1×109CFU/mL,例如1×109CFU/mL、3×109CFU/mL、5×109CFU/mL、8×109CFU/mL、1×1010CFU/mL、5×1010CFU/mL、1×1011CFU/mL、1×1012CFU/mL等;上述各项数值范围内的其他具体点值均可选择,在此便不再一一赘述。
优选地,所述嗜酸乳杆菌Lactobacillus acidophilus LA85菌株与长双歧杆菌Bifidobacterium longum BL21菌株的活菌数之比为1:5-5:1,例如1:5、1:4、1:3、1:2、1:1、2:1、3:1、4:1、5:1等,上述数值范围内的其他具体点值均可选择,在此便不再一一赘述。
当嗜酸乳杆菌LA85菌株和长双歧杆菌BL21菌株以上述特定的配比方式进行复合时,其在改善慢性酒精诱导的肝疾病的效果上更加显著,即在减少慢性酒精诱导的肝脂积累和肝损伤、调节慢性酒精诱导的肝疾病小鼠的肝脂质代谢水平、减轻慢性酒精诱导的肝疾病小鼠肝细胞凋亡程度方面的效果更加显著。
优选地,所述益生菌剂的剂型包括冻干粉剂、胶囊剂、片剂或颗粒剂。
本发明所涉及的益生菌剂的剂型不受限制,包括最常用的冻干粉剂,或进一步制得的胶囊剂、片剂或颗粒剂。其中冻干粉剂示例性地可以采用如下方法制得:
将嗜酸乳杆菌LA85菌株和长双歧杆菌BL21菌株分别接种于培养基中进行培养,得到培养液;培养液离心,得到菌体;菌体用冻干保护剂重悬,得到重悬液;重悬液冻干,即得,然后按比例将二者搭配。
优选地,所述培养基包括MRS培养基。
优选地,所述MRS培养基以浓度计包括:蛋白胨8-12g/L、牛肉膏8-12g/L、葡萄糖15-25g/L、乙酸钠1-3g/L、酵母粉3-7g/L、柠檬酸氢二铵1-3g/L、K2PO4·3H2O 2-3g/L、MgSO4·7H2O 0.05-0.2g/L、MnSO4 0.01-0.1g/L、吐温80 0.5-2mL/L、半胱氨酸盐酸盐0.1-1g/L。
优选地,所述冻干采用真空冷冻法。
优选地,所述改善慢性酒精诱导的肝疾病的益生菌剂还包括冻干保护剂和/或功能助剂。
优选地,所述冻干保护剂包括脱脂乳、明胶、糊精、阿拉伯胶、右旋糖酐、藻胶钠、聚乙烯吡咯烷酮、蔗糖、乳糖、海藻糖、山梨醇或木糖醇中的任意一种或至少两种的组合。
优选地,所述功能助剂包括低聚果糖、低聚半乳糖、低聚木糖、低聚异麦芽糖、大豆低聚糖、菊粉、螺旋藻、节旋藻、云芝多糖、水苏糖、聚葡萄糖、α-乳淸蛋白或乳铁蛋白中的任意一种或至少两种的组合。
上述功能助剂即益生元能够配合菌株发挥作用,从而进一步地提高益生菌剂在改善慢性酒精诱导的肝疾病上的效果。
第二方面,本发明提供第一方面所述的改善慢性酒精诱导的肝疾病的益生菌剂在制备预防、缓解或治疗慢性酒精性肝病的药物中的应用。
第三方面,本发明提供第一方面所述的改善慢性酒精诱导的肝疾病的益生菌剂在制备预防、缓解或治疗酒精性肝炎、酒精性肝纤维化或肝硬化的药物中的应用。
第四方面,本发明提供第一方面所述的改善慢性酒精诱导的肝疾病的益生菌剂在制备酒精诱导的肝细胞凋亡抑制剂中的应用。
由于第一方面所述的改善慢性酒精诱导的肝疾病的益生菌剂能够干预促使Bcl-2水平的升高和Bax的降低,从而增强肝细胞对凋亡的抵抗性,可以显著减轻酒精诱导的肝细胞凋亡,因此该益生菌剂还能够单纯地作为一种酒精诱导肝细胞凋亡的抑制剂,在肝细胞凋亡代谢活动的基础研究中发挥作用。
相对于现有技术,本发明具有以下有益效果:
本发明创造性地将嗜酸乳杆菌LA85菌株和长双歧杆菌BL21菌株进行复配,发现两者能够相互配合、相互促进,在改善慢性酒精诱导的肝疾病的效果上协同增效,具体表现在:能够在减少慢性酒精诱导的肝脂积累和肝损伤上协同增效;能够在调节慢性酒精诱导的肝疾病小鼠的肝脂质代谢水平上协同增效;能够在减轻慢性酒精诱导的肝疾病小鼠肝细胞凋亡程度上协同增效。相较于单菌使用,两种的组合方式在上述功效上更加显著,获得了本领域技术人员难以预见的技术效果,为改善慢性酒精诱导的肝疾病提供了新的策略,可以用于制备预防、缓解或治疗慢性酒精诱导的肝疾病的相关产品,具有广泛的应用前景。
附图说明
图1是各自小鼠血浆中丙氨酸氨基转移酶(ALT)水平的统计结果图;
图2是各自小鼠血浆中天冬氨酸氨基转移酶(AST)水平的统计结果图;
图3是各自小鼠肝甘油三酯含量的统计结果图;
图4是各自小鼠血浆游离脂肪酸含量的统计结果图;
图5是各自小鼠肝脏游离脂肪酸含量的统计结果图;
图6是各自小鼠肝脏组织中TUNEL阳性细胞比例的统计结果图;
图7是各自小鼠肝细胞中Bcl-2蛋白表达的统计结果图;
图8是各自小鼠肝细胞中Bax蛋白表达的统计结果图;
图9是各自小鼠AMPK的肝磷酸化水平的统计结果图;
图10是各自小鼠ACC的肝磷酸化水平的统计结果图;
图11是各自小鼠的肝脏SREBP-1c蛋白水平、PPAR-αmRNA水平、PGC-1αmRNA水平、SCD-1mRNA水平的统计结果图。
具体实施方式
下面通过具体实施方式来进一步说明本发明的技术方案。本领域技术人员应该明了,所述实施例仅仅是帮助理解本发明,不应视为对本发明的具体限制。
下述实施例中涉及的蛋白胨、牛肉膏、葡萄糖、乙酸钠、酵母粉、柠檬酸氢二铵、K2PO4·3H2O、MgSO4·7H2O、MnSO4、吐温80和半胱氨酸盐酸盐购自国药集团化学试剂有限公司。
下述实施例中涉及的培养基如下:
MRS培养基(g/L):蛋白胨10g/L、牛肉膏10g/L、葡萄糖25g/L、乙酸钠2g/L、酵母粉5g/L、柠檬酸氢二铵2g/L、K2PO4·3H2O 2.6g/L、MgSO4·7H2O 0.1g/L、MnSO4 0.05g/L、吐温80 1mL/L、半胱氨酸氨酸盐0.5g/L。
下述实施例所涉及的长双歧杆菌命名为长双歧杆菌Bifidobacterium longumBL21菌株,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,保藏时间为2015年01月27日,保藏编号为CGMCC 10452,地址为:北京市朝阳区北辰西路1号院3号。
下述实施例所涉及的嗜酸乳杆菌命名为嗜酸乳杆菌Lactobacillus acidophilusLA85菌株,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心,保藏时间为2020年07月20日,保藏编号为CGMCC 1.12735,地址为:北京市朝阳区北辰西路1号院3号。
下述实施例中涉及的菌悬液:将各菌株分别接种于脱脂乳中,37℃下培养18h进行活化,得到活化液;将活化液按2%(v/v)的接种量接种于MRS液体培养基中,37℃下培养18h,得到菌液;稀释即得。
实施例
本实施例探究益生菌剂对慢性酒精诱导的肝疾病小鼠肝脂积累和肝损伤、肝脂质代谢、酒精诱导的肝细胞凋亡的影响:
(1)动物分组和建立慢性酒精诱导的肝疾病小鼠模型:
将雄性C57BL/6N小鼠随机分为5组,每组10只,分别为正常对照组(CTL组)、饮酒模型组(AF组)、嗜酸乳杆菌LA85组(LA85组)、长双歧杆菌BL21组(BL21组)和复合益生菌组(LA85+BL21,活菌数比例1:1)。
所有小鼠均无特定病原体。将小鼠饲养于笼内,环境清洁安静,温度23-25℃,湿度50-70%。雄性C57BL/6小鼠由上海实验动物中心提供。所有涉及小鼠的程序均符合上海市实验动物护理与动物实验中心提供的指导原则(许可证号2022122008)。小鼠适应性饲养1周后,分为5组,饮酒组和益生菌干预组使用流质饲料(该饲料中的能量:17%为蛋白质,40%为玉米油,7.5%为碳水化合物,35.5%为酒精)及其对照组(等热量麦芽糖糊精),益生菌干预组干预剂量为1×109CFU/天/只,干预时间为4周。采集血浆和组织样本进行分析。
(2)生化分析:为了评估肝损伤,测量了肝酶的水平,分别使用ALT和AST检测试剂盒(武汉纯度生物)测量了血浆丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)。
统计结果如图1和图2所示(与AF组对比,**表示p<0.01),对比对照组,AF组小鼠由于酒精暴露,显著提高了血浆ALT和AST水平,而益生菌干预可以显著逆转这种情况,尤其是BL21+LA85复合组,基本能够恢复到正常水平,减轻了慢性酒精诱导的肝脏损伤。
(3)脂肪肝评估:为了评价益生菌干预对慢性酒精诱导的肝脂积累的影响,检测了肝甘油三酯、血浆游离脂肪酸、肝脏游离脂肪酸的水平,肝甘油三酯(TG)检测使用甘油三酯检测试剂盒(武汉纯度生物),游离脂肪酸(FFA)检测使用试剂盒(武汉纯度生物)。
统计结果如图3、图4、图5所示(与AF组对比,**表示p<0.01),与对照组小鼠相比,AF组小鼠的肝甘油三酯含量显著升高,而益生菌干预后,显著减少了慢性酒精诱导的肝甘油三酯积累,并且复合菌组效果最好。此外,长期接触酒精会增加血浆和肝脏FFA的水平,而益生菌干预可以阻止这种趋势,尤其是复合益生菌组,即益生菌干预可以有效减少慢性酒精诱导的肝脂积累和肝损伤。
(4)肝细胞凋亡分析:通过TUNEL染色(TUNEL法是分子生物学与形态学相结合的研究方法,常用于细胞凋亡研究。对完整单个凋亡细胞核或凋亡小体进行原位染色,能准确地反应细胞凋亡典型生物化学和形态特征,由于正常的或正在增殖的细胞几乎没有DNA的断裂,因而没有3-OH形成,很少能够被染色。TUNEL法可用于石蜡包埋组织切片、冰冻组织切片、培养的细胞和从组织中分离的细胞的形态测定,并可检测出极少量的凋亡细胞)评估肝脏细胞凋亡。通过计算5个随机选择的视图字段进行定量的TUNEL阳性细胞,并表示为TUNEL阳性细胞/500个细胞。根据制造商的说明,用ApopTag过氧化物酶原位凋亡检测,对福尔马林固定石蜡肝切片进行染色,用于末端脱氧核苷酸转移酶介导的dUTP镍端标记。同时使用Western blot法检测抗凋亡和促凋亡蛋白表达情况来评估慢性酒精暴露诱导小鼠肝细胞凋亡程度。
统计结果如图6、图7、图8所示(与AF组对比,**表示p<0.01),与对照组相比,AF组酒精喂养小鼠的肝脏组织中TUNEL阳性细胞显著增加,益生菌干预后显著减少了肝组织中的TUNEL阳性细胞,尤其是复合益生菌组;通过检测抗凋亡和促凋亡蛋白表达情况来评估慢性酒精暴露诱导小鼠肝细胞凋亡程度,与AF组小鼠相比,益生菌干预组小鼠的Bcl-2蛋白表达显著提高,尤其是复合组,基本与正常水平持平,另外AF组小鼠的Bax蛋白表达水平显著提高,Bax基因是人体最主要的凋亡基因,细胞凋亡促进基因,而益生菌干预可以减轻这种凋亡信号表达异常升高的情况,尤其是复合益生菌组。即复合益生菌组的干预促使Bcl-2水平的升高和Bax的降低,从而增强机体细胞对凋亡的抵抗性,可以显著减轻酒精诱导的肝细胞凋亡。
(5)对肝脏AMPK信号传导的影响:通过测定慢性酒精干预小鼠AMPK的肝磷酸化水平,来评估益生菌干预对酒精诱导肝疾病小鼠肝脏AMPK信号传导的影响情况。使用Westernblot法检测AMPK和ACC磷酸化水平,并通过密度分析进行量化,以测定益生菌干预对接触慢性酒精的小鼠肝脏蛋白激酶腺苷酸活化蛋白激酶(AMPK)和乙酰辅酶A羧化酶(ACC)磷酸化的影响。
统计结果如图9、图10所示(与AF组对比,**表示p<0.01),与对照组相比,AF组小鼠酒精诱导显著降低了肝脏AMPK磷酸化,而益生菌干预后这种情况得到有效的逆转,尤其是复合益生菌干预组。已有研究表示,AMPK(p-AMPKα)的Thr-172磷酸化可以激活AMPK,在肝脂质代谢中起着关键作用。AMPK激活通过磷酸化来降低ACC活性,ACC是脂肪合成的限速酶,它催化脂肪酸生物合成的第一步反应,生成丙二酰辅酶A,然后进一步形成长链脂肪酸,最后合成三酰甘油和磷脂。与对照小鼠相比,AF组小鼠肝脏的ACC磷酸化水平明显较低,而益生菌干预可以有效的抑制这种降低,即提高ACC磷酸化表达水平,从而降低ACC活性,肝脏脂肪合成减少,尤其是复合益生菌组。即复合益生菌能够参与肝脏的AMPK信号传导,有效调节肝脏脂代谢。
(6)对接触慢性酒精的小鼠肝脏表达的影响:通过检测肝脏SREBP-1c蛋白和PPAR-α转录水平,评估益生菌干预对慢性酒精暴露诱导的脂质合成和清除的影响。用Westernblot测定SREBP-1c的蛋白质水平,并通过密度分析量化蛋白质带密度。用实时RT-PCR分析了PPAR-α、过氧化物酶体增殖物激活受体-γ共激活因子-1α(PGC-1a)和硬脂酰辅酶A脱脂酶-1(SCD-1)的肝脏mRNA水平。
统计结果如图11所示(与AF组对比,**表示p<0.01),与对照组相比,AF组小鼠在慢性酒精暴露后会显著增加肝脏SREBP-1c、SCD-1蛋白的表达,显著降低肝脏PPAR-α和PGC-1αmRNA表达。SREBP-1和PPAR-α是调节肝脂质代谢的转录因子,在新脂肪生成和脂肪酸氧化中起着核心作用,SCD-1是负责脂肪酸合成的关键酶,也是SREBP-1的靶标。经益生菌干预后,这种异常升高情况有所逆转,尤其是益生菌组合组。总的来说,益生菌通过下调SREBP-1、上调PPAR-α来参与肝脏脂代谢,可以有效积极调节SRBEP-1c和PPAR-α介导的慢性酒精诱导的脂肪生成和脂肪清除的变化,尤其是复合益生菌组的效果最佳。
申请人声明,本发明通过上述实施例来说明本发明的一种改善慢性酒精诱导的肝疾病的益生菌剂及其应用,但本发明并不局限于上述实施例,即不意味着本发明必须依赖上述实施例才能实施。所属技术领域的技术人员应该明了,对本发明的任何改进,对本发明产品各原料的等效替换及辅助成分的添加、具体方式的选择等,均落在本发明的保护范围和公开范围之内。
以上详细描述了本发明的优选实施方式,但是,本发明并不限于上述实施方式中的具体细节,在本发明的技术构思范围内,可以对本发明的技术方案进行多种简单变型,这些简单变型均属于本发明的保护范围。
另外需要说明的是,在上述具体实施方式中所描述的各个具体技术特征,在不矛盾的情况下,可以通过任何合适的方式进行组合,为了避免不必要的重复,本发明对各种可能的组合方式不再另行说明。
Claims (10)
1.一种改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,所述改善慢性酒精诱导的肝疾病的益生菌剂中的菌株包括保藏编号为CGMCC No.1.12735的嗜酸乳杆菌Lactobacillus acidophilus LA85菌株和保藏编号为CGMCCNo.10452的长双歧杆菌Bifidobacterium longum BL21菌株。
2.根据权利要求1所述的改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,在益生菌剂中,所述嗜酸乳杆菌Lactobacillus acidophilus LA85菌株的活菌数不低于1×109CFU/mL;所述长双歧杆菌Bifidobacterium longum BL21菌株的活菌数不低于1×109CFU/mL。
3.根据权利要求1或2所述的改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,所述嗜酸乳杆菌Lactobacillus acidophilus LA85菌株与长双歧杆菌Bifidobacteriumlongum BL21菌株的活菌数之比为1:5-5:1。
4.根据权利要求1-3中任一项所述的改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,所述益生菌剂的剂型包括冻干粉剂、胶囊剂、片剂或颗粒剂。
5.根据权利要求1-4中任一项所述的改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,所述改善慢性酒精诱导的肝疾病的益生菌剂还包括冻干保护剂和/或功能助剂。
6.根据权利要求5所述的改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,所述冻干保护剂包括脱脂乳、明胶、糊精、阿拉伯胶、右旋糖酐、藻胶钠、聚乙烯吡咯烷酮、蔗糖、乳糖、海藻糖、山梨醇或木糖醇中的任意一种或至少两种的组合。
7.根据权利要求5所述的改善慢性酒精诱导的肝疾病的益生菌剂,其特征在于,所述功能助剂包括低聚果糖、低聚半乳糖、低聚木糖、低聚异麦芽糖、大豆低聚糖、菊粉、螺旋藻、节旋藻、云芝多糖、水苏糖、聚葡萄糖、α-乳淸蛋白或乳铁蛋白中的任意一种或至少两种的组合。
8.权利要求1-7中任一项所述的改善慢性酒精诱导的肝疾病的益生菌剂在制备预防、缓解或治疗慢性酒精性肝病的药物中的应用。
9.权利要求1-7中任一项所述的改善慢性酒精诱导的肝疾病的益生菌剂在制备预防、缓解或治疗酒精性肝炎、酒精性肝纤维化或肝硬化的药物中的应用。
10.权利要求1-7中任一项所述的改善慢性酒精诱导的肝疾病的益生菌剂在制备酒精诱导的肝细胞凋亡抑制剂中的应用。
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WO2019212997A1 (en) * | 2018-04-30 | 2019-11-07 | Rejuvenation Therapeutics | Compositions and methods for biosynthetic preparation of urolithin compounds and use thereof |
CN112914103A (zh) * | 2021-02-22 | 2021-06-08 | 漯河微康生物科技有限公司 | 一种抗幽门螺杆菌感染的益生菌组合物及其应用 |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108208853A (zh) * | 2018-01-04 | 2018-06-29 | 山东凤凰生物有限公司 | 一种解酒护肝益生菌低聚肽复合制剂及制备方法 |
WO2019212997A1 (en) * | 2018-04-30 | 2019-11-07 | Rejuvenation Therapeutics | Compositions and methods for biosynthetic preparation of urolithin compounds and use thereof |
CN112914103A (zh) * | 2021-02-22 | 2021-06-08 | 漯河微康生物科技有限公司 | 一种抗幽门螺杆菌感染的益生菌组合物及其应用 |
Non-Patent Citations (2)
Title |
---|
TAO WU: "Bifidobacterium longum subsp. longum Remodeled Roseburia and Phosphatidylserine Levels and Ameliorated Intestinal Disorders and liver Metabolic Abnormalities Induced by High-Fat Diet", AGRICULTURAL AND FOOD CHEMISTRY, pages 4632 - 4640 * |
赵琳 等人: "嗜酸乳杆菌LA85对小鼠免疫机能的研究", 中国食物与营养, pages 65 - 69 * |
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