CN114191490A - 一种发酵乳杆菌及其产物在抗氧化和抗肿瘤中的应用 - Google Patents
一种发酵乳杆菌及其产物在抗氧化和抗肿瘤中的应用 Download PDFInfo
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Abstract
本发明公开了一种发酵乳杆菌及其产物在抗氧化和抗肿瘤中的应用,属于天然产物应用技术领域。具体公开了发酵乳杆菌及其产物在制备抗氧化药物、食品或保健品中的应用,以及发酵乳杆菌及其产物在制备抗肿瘤药物、食品或保健品中的应用,所述发酵乳杆菌为发酵乳杆菌CQPC04,保藏编号为CGMCC NO.14493;所述发酵乳杆菌产物是经发酵乳杆菌CQPC04发酵葡萄皮获取。实验验证显示,经过发酵乳杆菌CQPC04发酵的葡萄皮,获取的发酵产物相比其他菌种发酵或者直接化学试剂提取的化学成分,具有显著提高抗氧化和抑制肝癌细胞增殖的作用。
Description
技术领域
本发明涉及天然产物应用技术领域,特别是涉及一种发酵乳杆菌及其产物在抗氧化和抗肿瘤中的应用。
背景技术
许多自由基是通过呼吸和新陈代谢产生的。在正常生理条件下,自由基的产生和抗氧化系统在体内处于动态平衡状态,当自由基的积累超过抗氧化系统的保护能力时,平衡被打破,表现为氧化应激。研究表明,氧化应激可损伤细胞中的各种大分子(如多糖、蛋白质、脂质和DNA),氧化应激还可刺激参与细胞凋亡、炎症反应、细胞功能改变的信号通路,并最终导致疾病的病理变化,如帕金森病、阿尔茨海默病和癌症。因此,提高抗氧化能力和适当摄入抗氧化食物对健康很重要。研究表明,南瓜多糖可以减轻肝细胞的氧化损伤,类黄酮可以抑制H2O2氧化损伤引起的溶血,减少过氧化物的产生,增加过氧化物酶的活性。此外,多酚可以改善帕金森病大鼠的氧化应激反应。
肝脏是体内主要的代谢器官。肝细胞癌(HCC)占原发性肝癌90%以上,是最常见的恶性肿瘤之一。环境因素与HCC发病机制密切相关,如乙型和丙型肝炎病毒感染、饮用污染的水、寄生虫病和化学物质。由于肝癌的发生具有隐匿性,大多数患者在诊断时已经进入中晚期。由于肝脏有丰富的血液供应,癌细胞很容易转移到其他器官,使得HCC很难治疗。
癌细胞和组织的生长和转移需要大量的能量供应。癌细胞ATP的产生依赖于发达的线粒体和糖酵解系统,在任何环境下都能保持高能量的产生状态。大多数抗癌药物通过作用于线粒体引起功能损伤而发挥抗癌作用。相关研究表明,沉默相关癌基因或蛋白表达可干扰线粒体活性,激活死亡受体活性,从而促进细胞凋亡或自噬凋亡。
葡萄皮含有许多活性成分,包括多酚、纤维素和蛋白质,如白藜芦醇、鞣酸、儿茶素、槲皮素和花青素,具有很强的抗氧化、抗突变、抗菌、抗炎、心血管保护等作用。随着葡萄产量的增加和葡萄产业的快速发展,我国葡萄渣年产量也在快速增长。只有25%的渣土可以回收再利用,大部分用作饲料或肥料或被丢弃。最近的深入研究已从葡萄皮残渣中鉴定出10多种单体酚,包括儿茶素、槲皮素、没食子酸和绿原酸。但是对于如何高效的利用葡萄皮中的有效成分用于抗氧化或者抗肿瘤作用还没有相关的报道。
发明内容
本发明的目的是提供一种发酵乳杆菌及其产物在抗氧化和抗肿瘤中的应用,以解决上述现有技术存在的问题,经过发酵乳杆菌CQPC04发酵的葡萄皮,获取的发酵产物具有较高的抗氧化和抑制肝癌细胞增殖的作用。
为实现上述目的,本发明提供了如下方案:
本发明提供一种发酵乳杆菌及其产物在制备抗氧化药物、食品或保健品中的应用,所述发酵乳杆菌为发酵乳杆菌(Lactobacillus fermentum)CQPC04,保藏编号为CGMCCNO.14493,保藏时间为2017年8月4日,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,保藏地址为北京市朝阳区北辰西路1号院3号中国科学院微生物研究所;所述发酵乳杆菌产物是经发酵乳杆菌CQPC04发酵葡萄皮获取。
本发明还提供一种发酵乳杆菌或其产物在制备抗肿瘤药物、食品或保健品中的应用,所述发酵乳杆菌为发酵乳杆菌CQPC04,保藏编号为CGMCC NO.14493;所述发酵乳杆菌产物是经发酵乳杆菌CQPC04发酵葡萄皮获取。
优选的是,所述发酵乳杆菌CQPC04发酵葡萄皮的方法包括以下步骤:
将葡萄皮干燥后,研磨成粉末,得到葡萄皮粉;
将所述葡萄皮粉和糖溶于超纯水中,接种所述发酵乳杆菌CQPC04,37℃发酵培养,获取发酵液。
优选的是,所述葡萄皮粉为500目。
优选的是,所述葡萄皮:糖:超纯水的配比为(8-15)g:1g:100mL。
优选的是,所述葡萄皮粉:所述发酵乳杆菌CQPC04的配比为(8-15)g:1mL,其中所述发酵乳杆菌CQPC04菌浓为1×107CFU/mL。
优选的是,所述肿瘤为肝癌。
优选的是,所述发酵乳杆菌及其产物降低H2O2诱导的氧化损伤。
优选的是,所述发酵乳杆菌及其产物促进细胞凋亡,抑制肿瘤细胞增殖。
本发明公开了以下技术效果:
本发明通过采用发酵乳杆菌CQPC04对葡萄皮进行发酵,所获取的发酵产物经实验验证发现,对293T细胞具有抗氧化作用,对HepG2细胞具有促凋亡作用。并且明显优于德氏乳杆菌保加利亚亚种发酵葡萄皮产物,以及化学方法乙醇提取葡萄皮获取的葡萄皮产物,因此,本发明提供了一种生物发酵方法获取的发酵乳杆菌或其产物方法,该方法简单,且不涉及大量化学试剂的使用,通过该方法获取的发酵产物具有显著的抗氧化和抑制肿瘤细胞增殖的作用,这为抗氧化和肿瘤治疗药物或食品(尤其是保健食品)的筛选提供了新的方向。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为各实验组的体外抗氧化能力;A:每个实验组在不同时间间隔(24、48、72、96、120h)对DPPH的抗性;B:每个实验组在不同时间间隔(24、48、72、96、120h)对ABTS的抗性;给出的值是平均标准偏差(N=3/组)。(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:乙醇提取的溶液)(*代表P<0.05,**代表0.05<P<0.01,***代表0.01<P<0.001,***代表0.001<P<0.0001);
图2为时间和H2O2浓度对细胞存活率的影响;
图3为葡萄皮发酵液对细胞生长的影响;A:葡萄皮发酵液处理48h对293T细胞生长和活力的影响;B:葡萄皮发酵液处理48h对HepG2细胞生长和活力的影响;(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液);
图4为处理葡萄皮发酵液48h对293T细胞活力(200×)的影响;活细胞用绿色荧光标记,死细胞用红色荧光标记;(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液);
图5为葡萄皮发酵液处理48h对HepG2细胞活力(200×)的影响;活细胞用绿色荧光标记,死细胞用红色荧光标记;(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液);
图6为SOD,GSH,CAT和GSH-Px在293细胞中的表达情况;用Duncan多重极差检验,同一组中不同字母的a-e均值差异显著(P<0.05);给出的值是平均标准偏差(N=3/组);(CF:LF-CQPC 04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液);
图7为293T细胞中SOD,GSH,CAT和GSH-Px蛋白的表达情况;给出的值是平均标准偏差(N=3/组);(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:乙醇提取的溶液)(*代表P<0.05,**代表0.05<P<0.01,***代表0.01<P<0.001,***代表0.001<P<0.0001);
图8为葡萄皮发酵液对HepG2细胞周期和细胞凋亡的影响;(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液);
图9为肝癌细胞中的基因表达;Duncan多重极差检验显示,同一组中不同字母的a-d均值差异显著(P<0.05);给出的值是平均标准偏差(N=3/组)(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液);
图10为Phospho-NF-κB(p65),Bax,Caspase-8和NF-κB(p65)蛋白在肝癌细胞中的表达;给出的值是平均标准偏差(N=3/组);(CF:LF-CPQC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:乙醇提取的溶液)(*代表P<0.05,**代表0.05<P<0.01,***代表0.01<P<0.001,***代表0.001<P<0.0001);
图11为葡萄皮发酵液中的多酚成分的标准色谱;1:表儿茶素没食子酸酯(ECG);2:香豆素;3:新绿原酸;4:芦丁;5:白藜芦醇;6:绿原酸;
图12为LF-CQPC04发酵葡萄皮所得发酵液(CF)的色谱图;
图13为德氏乳杆菌保加利亚亚种发酵葡萄皮所得发酵液(BF)的色谱图。
具体实施方式
现详细说明本发明的多种示例性实施方式,该详细说明不应认为是对本发明的限制,而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式,并非用于限制本发明。另外,对于本发明中的数值范围,应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明,否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料,但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入,用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时,以本说明书的内容为准。
在不背离本发明的范围或精神的情况下,可对本发明说明书的具体实施方式做多种改进和变化,这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见的。本申请说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等,均为开放性的用语,即意指包含但不限于。
实施例1
一、材料与方法
1、样品处理
将新鲜巨峰葡萄皮真空干燥,并通过500目筛研磨成粉末。发酵组:将10g葡萄皮粉和1g白糖溶于100mL超纯水中,然后将1mL 1×107CFU/mL发酵乳杆菌CQPC04(LF-CQPC04)接种于溶液中,菌株对照组:接种1mL 1×107CFU/mL德氏乳杆菌保加利亚亚种(中国微生物菌种保藏中心,保藏编号:CGMCC NO.1.16075)。该溶液在100rpm的摇床中于37℃发酵。24、48、72、96和120h后,溶液以12000rpm离心10min,然后收集上清液并在-80℃下储存。空白对照组,将10g葡萄皮粉末溶解在100mL 60%乙醇溶液中,在70℃下水浴4h。通过离心丢弃沉淀物,并将溶液旋转蒸发(中国河南郑州郑州长城科技工贸有限公司R-1001VN)以除去(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液)。
2、实验方法
2.1体外抗氧化
2,2-二苯基-1-吡啶并肼基(DPPH)自由基清除分析(索拉博生命科学,北京,中国)按照Thaipong等人所述的方法进行。将发酵样品(24、48、72、96和120h)加入0.1mM/mLDPPH,溶解在甲醇(0.1mL)溶液中并充分混合。室温下在黑暗中孵育30min后,在570nm测量吸光度。根据Roberta等人所述的方法制备2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)(索莱宝生命科学,北京,中国)工作液。简而言之,将20μL发酵样品溶液和200μLABTS工作溶液在孔混合均匀,在吸光度720nm处用微孔板读数器测量(Thermo FisherScientific,New York,USA)。
2.2细胞氧化损伤的诱导
将人胚胎肾(293T)细胞(293T细胞系购自中国科学院细胞库(上海,中国))接种在1×106的96孔板中,然后在恒温培养箱(thermo371,Thermo Fisher Scientific,NewYork,USA)中,于37℃和5%CO2下与DMEM(H)(索莱宝生命科学,北京,中国)一起培养24h。细胞完全粘附后,吸取培养液,用PBS缓冲溶液洗涤一次。将293T细胞随机分为8组。空白组含有200μL细胞培养基,但未接种细胞。对照组接种细胞,加200μL细胞培养液,不加H2O2。实验组在氧化损伤模型的200μL细胞培养基中包含不同浓度的H2O2(0、100、200、300、400、500、600和700μmol/L),每组重复3次。在培养细胞1、2和3h后,丢弃原始培养液,加入100μL新培养液,并使用CCK-8方法(索莱宝生命科学,中国北京)测量细胞活力。
2.3药物剂量浓度筛选
将293T细胞和人肝癌(HepG2)细胞(HepG2细胞系购自中国科学院细胞库(上海,中国))接种在1×106的96孔板中,并在37℃和5%CO2的恒温培养箱中培养24h,DMEM(H)和DMEM(L)(索莱宝生命科学,北京,中国)。细胞完全粘附后,吸取培养液并用PBS洗涤一次。将HepG2和293T细胞随机分为8组。空白组含有200μL培养基,但未接种细胞。对照组接种细胞,包含200μL细胞培养基,但不包含提取物。实验组接受不同提取物浓度(0、100、200、300、400、500、600、700μmol/L)和200μL细胞培养基进行处理,每组3次重复。细胞培养48h后,弃去原始培养液,加入100μL新培养液,用CCK-8法测定细胞活力。
2.4细胞指数的测量
使用超声波细胞破碎器(超声均质器Scientz-IID,宁波新芝生物科技股份有限公司,中国浙江)破碎细胞样品。用试剂盒(南京建成生物工程研究所)测定乳酸脱氢酶、谷胱甘肽、氧化型谷胱甘肽、过氧化氢酶、超氧化物歧化酶、过氧化谷胱甘肽、总抗氧化能力、一氧化氮和丙二醛的含量。
2.5观察细胞活力
将5×106个细胞接种在6孔细胞平板上,培养48h(细胞汇合度达到80%以上),然后进行相应的药物治疗。使用钙黄绿素-AM/PI双染色试剂盒通过倒置荧光显微镜观察细胞死亡率。活细胞用绿色荧光标记,死细胞用红色荧光标记。
2.6细胞周期和凋亡的测量
细胞样品用0.25%乙二胺四乙酸-胰蛋白酶处理(索莱宝生命科学,中国北京),并在4℃下以1000rpm离心10min,然后除去上清液,用已经在4℃预冷却的PBS洗涤,并再次离心10min。使用细胞周期和凋亡分析试剂盒(翌圣生物科技(上海)股份有限公司,上海,中国)通过流式细胞术检测细胞周期。采用Annexin V-FITC/PI凋亡检测试剂盒(翌圣生物科技(上海)股份有限公司,中国上海),通过流式细胞仪(AccuriC6,BD Biosciences,SanJose,CA,USA)检测细胞凋亡。
2.7逆转录-定量聚合酶链反应
使用检测试剂盒提取总RNA(百迈客;中国北京)。使用显微分光光度计(Nano-300,杭州奥盛仪器有限公司,中国浙江杭州)测定RNA浓度和纯度。RNA反转录成cDNA第一步为利用cDNA合成试剂盒(Thermo Fisher Scientific,Baltics,USA)中逆转录酶将RNA反转录成cDNA。使用HieffTM qPCRGreen Master Mix(High Rox Plus)(翌圣生物科技(上海)股份有限公司,中国上海)和第一步实时PCR系统(Thermo Fisher Scientific,美国纽约)测量产物评估靶基因表达水平。用肌动蛋白标准化靶基因表达,并用2-ΔΔCt方法计算。表1列出了使用的RT-qPCR引物。
表1 qPCR引物序列
使用以下扩增条件进行RT-qPCR:在95℃预变性3min,随后在95℃变性30s,X℃退火30s(X表示通过梯度PCR确定的退火温度(A200梯度热循环,浙江龙基因科学仪器有限公司,中国浙江杭州))(表1),72℃延伸30s,40个循环。
2.8蛋白质印迹分析
使用200μL的RIPA裂解液和2μL的苯甲基磺酰氟(PMSF)(柏奥易杰(北京)科技有限公司,中国北京)从细胞中提取总蛋白。将蛋白质在冰上提取30min,并在100℃下煮沸10min使其变性。使用Bradford法蛋白浓度测定试剂盒(Bio-Rad Laboratories,Hercules,CA,USA)测定提取的蛋白质浓度。使用12%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离蛋白质(30μg),并转移至硝酸纤维素膜。将这些膜在5%脱脂牛奶溶液中密封2h,然后用TBST 1×洗涤5次,并与抗SOD、CAT、GSH、GSH-Px、Caspase-8、Phospho-NF-κB(p65)、NF-κB(p65)、Bax和β-肌动蛋白的单克隆抗体(表达载体,Thermo Fisher Scientific,RockFord,USA)一起在4℃孵育过夜最后,将印迹与增强化学发光(ECL)辣根过氧化物酶底物一起孵育(索莱宝生命科学,中国北京),并使用Tanon 5200发光成像分析系统(天能科技有限公司,中国上海)对表达产物进行成像。使用ImageJ 1.44软件进行蛋白质表达的半定量分析。
2.9高效液相色谱分析
称取对羟基肉桂酸、新绿原酸、绿原酸、芦丁、虎杖苷、迷迭香酸和表儿茶素没食子酸酯的标准品(上海源叶生物技术有限公司),用甲醇配制0.1mg/mL溶液。用超高效液相色谱C18柱(Thermo Scientific,Bellefonte,PA.),用甲醇-水(1∶1,V/V)洗脱,并通过0.22μL过滤器过滤。使用以下色谱条件检测发酵组分(UltiMate3000高效液相色谱仪,ThermoFisher Scientific,USA):Accucore C18柱(4.6mm×150mm,2.6μm,Thermo FisherScientific),流速0.5mL/min,检测波长285nm,注射体积10L,柱温30℃,收集时间75min,流动相A为乙腈,流动相B为0.1%乙酸水溶液。
2.10统计分析
数据采用SPSS 17.0和GraphPad Prism统计软件进行统计分析。实验结果用平均标准差(SD)表示。组间比较采用单因素方差分析或t检验。P<0.05表明有统计学意义的差异。所有实验重复三次。
二、结果
1、抗氧化活性
葡萄皮发酵具有更高的抗氧化能力和更好的时间依赖性(图1)。为了确定发酵液的抗氧化能力,测定了发酵液的DPPH和ABTS自由基清除活性。LP-CQPC04发酵液在72、96和120h的DPPH自由基清除率分别为97.54%、92.86%和92.69%。发酵液在72、96和120h的ABTS自由基清除率分别为81.00%、93.60%和92.43%。德氏乳杆菌保加利亚亚种发酵液对DPPH和ABTS的抗性在120h最高,分别为86.68%和68.54%。乙醇提取对DPPH和ABTS的抗性率分别为54.53%和57.98%。
2、H2O2诱导人胚肾(293)细胞氧化损伤模型
不同H2O2浓度对293T细胞生长的抑制作用不同,细胞存活率随浓度增加而降低(图2)。与对照组相比,H2O2处理显著抑制293T细胞的存活,且抑制作用随时间、浓度、作用时间和浓度依赖性而增强。用100μmol/L H2O2处理2h,细胞活力显著下降,IC50为54.67±3.16%,与对照组(95.70±5.32%)差异显著。如果浓度太低,细胞损伤会最小;如果浓度过高,细胞死亡过多。因此,氧化损伤模型中使用的H2O2浓度为100μmol/L,作用时间为2h。
3、葡萄皮发酵液对人胚肾(293T)细胞和人肝癌(HepG2)细胞生长的影响
与空白对照组相比,将发酵液应用于293T细胞48h促进了细胞增殖(图3A)。当CF组浓度超过400μmol/L时,随着浓度的增加,细胞增殖明显受到抑制,BF和WE细胞增殖不明显。因此,300μmol/L的CF浓度被用作氧化损伤后续实验的推荐剂量。48h后,将葡萄皮发酵液应用于人肝癌HepG2细胞(图3B),并与空白对照组进行比较。随着时间的推移,细胞生长在这两个时期都受到明显抑制。这种增长是长期的,并且取决于时间。当CF浓度为150μmol/L时,作用时间为48h,细胞活力为55.42±1.2%。当BF浓度为200μmol/L时,作用时间为48h,细胞活力为53.20±2.6%。当WE浓度为300μmol/L时,作用时间为48h,细胞活力为44.48±1.5%。
4、葡萄皮发酵液对细胞活力的影响
在倒置荧光显微镜(Olympus,Tokyo,Japan)下观察细胞活力。正常的293T细胞数量众多,饱满,呈多角形,胞浆丰富,周围有假足交错排列,荧光染色可见大量活细胞(图4)。H2O2诱导模型组细胞严重萎缩,数量减少,损伤严重,细胞死亡率增加。CF治疗组活细胞数量增加,状态良好;在H2O2诱导的氧化后,CF似乎更好地保护细胞免于凋亡,增加细胞活力并减少损伤。BF和WE也保护细胞免受氧化损伤,但效果不如CF。在对照组中,HepG2细胞呈不规则多边形或长梭形,活细胞染色清晰(图5)。CF处理后,细胞体积缩小,死亡细胞比例明显增加;BF和WE治疗组的细胞凋亡也没有CF组严重。
5、人胚肾细胞中LDH,GSH,GSSG,GSH-Px,CAT,MDA,SOD,T-AOC,和NO含量测定
正常组GSH,GSH-Px,CAT,SOD和T-AOC含量最高,CF组GSH,GSH-Px和CAT含量明显高于WE组(表2)(P<0.05)。而正常细胞组LDH、MDA和NO含量最低,CF组NO含量明显低于对照组和WE组(P<0.05)。LDH含量也明显低于BF组和WE组,与正常组相似。MDA含量在CF组、BF组和WE组之间差异不显著,但MDA含量显著高于对照组(P<0.05)。氧化损伤模型中GSSG相对表达上调,对照组总-GSH+GSSG低于正常及其他实验治疗组,对照组GSH/GSSG比率下调(表3)(p<0.05)。
表2 293T细胞中LDH、GSH-Px、CAT、MDA、SOD、T-AOC和NO含量测定结果
Duncan多重极差检验显示,同一列中不同字母的a-c均值差异显著(P<0.05)。给出的值是平均标准偏差(N=3/组)。(CF:LF-CQPC 04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液;WE:用乙醇提取的溶液)。
表3 293T细胞中谷胱甘肽、GSSG和谷胱甘肽/GSSG含量
Duncan多重极差检验显示,同一列中不同字母的a-c均值差异显著(P<0.05)。给出的值是平均标准偏差(N=3/组)。(CF:LF-CQPC04发酵的发酵液;BF:德氏乳杆菌保加利亚亚种发酵的发酵液。人;WE:用乙醇提取的溶液)。
6、人胚肾细胞中SOD,GSH,CAT和GSH-Px mRNA和蛋白表达
SOD,GSH,CAT,and GSH-Px的mRNA表达在正常组最高,在对照组最低(图6)。然而,CF和WE抗氧化保护后,mRNA表达水平明显高于对照组。CF组SOD,GSH,CAT and GSH-Px的mRNA表达高于WE组和BF组。图7显示了通过蛋白质印迹分析测定的SOD,CAT,GSH,and GSH-Px蛋白水平。与正常组相比,模型组SOD,CAT,GSH,and GSH-Px蛋白水平明显降低。然而,与模型组相比,经LF-CQPC04处理的葡萄发酵后,SOD,CAT,GSH,and GSH-Px的蛋白水平显著升高。BF和WE还显著提高SOD,CAT,GSH,and GSH-Px蛋白水平。
7、葡萄皮发酵液对人肝癌细胞周期和凋亡的影响
如图8所示,正常的HepG2细胞处于G0/G1期达到73.2%,可转化为G2期(23.6%),细胞增殖正常。CF处理HepG2细胞48小时后,S/G2期延长(53.4%),大部分细胞仍处于S期,不能转化为G2期。在WE组和BF组中显示了相同的结果。凋亡结果表明,经发酵液处理的细胞凋亡和细胞碎片数量增加。与正常组相比,CF组、BF组、WE组的细胞活力分别为36.3%、51.3%和81.6%。CF治疗后细胞晚期凋亡数占63.7%,BF和WE分别为47.3%和17.1%。
8、人肝癌细胞相关基因和蛋白表达
如图9所示,与对照组相比,CF组细胞中Bcl-2、cox-2、PCNA、CD1、C-meyc、CDK4、NF-κB和pRb1的mRNA表达水平显著降低(P<0.05)。Caspase-3、Caspase-7、Caspase-8、Caspase-9、p53、TGF-β和p21的表达水平均高于正常组。与对照组相比,WE组和BF组还下调了Bcl-2、cox-2、PCNA、CD1、C-meyc、CDK4、NF-κB和pRb1的mRNA表达水平,上调了Caspase-3、Caspase-7、Caspase-8、Caspase-9、p53、TGF-β和p21的mRNA表达水平。图10显示了通过蛋白质印迹分析测定的phospho-p65,Bax,Caspase-8和NF-κB(p65)蛋白的水平。与正常组相比,CF治疗组phospho-p65,Bax和Caspase-8蛋白水平显著升高,NF-κB(p65)蛋白水平降低。BF和WE治疗组产生了相同的结果,phosphor-p65,Bax and Caspase8的蛋白水平显著增加,炎症因子NF-κB(p65)的蛋白表达水平降低。
9、高效液相色谱分析
如图11-图13显示了葡萄发酵液中通过高效液相色谱分离的多酚成分。如图12所示,CF中新绿原酸、芦丁、白藜芦醇的含量最高,分别达到37.276、29.930、28.533mAV*min,50.21min时的最高未知含量为56.85mAV*min(图12)。从分离结果来看,LF-CQPC04发酵的发酵液的总峰面积低于乙醇得到的葡萄皮提取物的总峰面积,CF发酵液分离的有效峰为57,乙醇提取物的有效峰为83。有效成分少于乙醇提取,BF发酵结果只积累了46个有效峰(图13)。通过标准对照比较,发酵液包括表儿茶素没食子酸酯、香豆素、新绿原酸、芦丁、白藜芦醇、绿原酸和迷迭香酸(图11)。
以上所述的实施例仅是对本发明的优选方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案做出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。
Claims (9)
1.一种发酵乳杆菌及其产物在制备抗氧化药物、食品或保健品中的应用,其特征在于,所述发酵乳杆菌为发酵乳杆菌CQPC04,保藏编号为CGMCC NO.14493;所述发酵乳杆菌产物是经发酵乳杆菌CQPC04发酵葡萄皮获取。
2.一种发酵乳杆菌及其产物在制备抗肿瘤药物、食品或保健品中的应用,其特征在于,所述发酵乳杆菌为发酵乳杆菌CQPC04,保藏编号为CGMCC NO.14493;所述发酵乳杆菌产物是经发酵乳杆菌CQPC04发酵葡萄皮获取。
3.如权利要求1或2所述的应用,其特征在于,所述发酵乳杆菌CQPC04发酵葡萄皮的方法包括以下步骤:
将葡萄皮干燥后,研磨成粉末,得到葡萄皮粉;
将所述葡萄皮粉和糖溶于超纯水中,接种所述发酵乳杆菌CQPC04,37℃发酵培养,获取发酵液。
4.如权利要求3所述的应用,其特征在于,所述葡萄皮粉为500目。
5.如权利要求3所述的应用,其特征在于,所述葡萄皮:糖:超纯水的配比为(8-15)g:1g:100mL。
6.如权利要求3所述的应用,其特征在于,所述葡萄皮粉:所述发酵乳杆菌CQPC04的配比为(8-15)g:1mL,其中所述发酵乳杆菌CQPC04菌浓为1×107CFU/mL。
7.如权利要求2所述的应用,其特征在于,所述肿瘤为肝癌。
8.如权利要求1所述的应用,其特征在于,所述发酵乳杆菌及其产物降低H2O2诱导的氧化损伤。
9.如权利要求2所述的应用,其特征在于,所述发酵乳杆菌及其产物促进细胞凋亡,抑制肿瘤细胞增殖。
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114642686A (zh) * | 2022-04-29 | 2022-06-21 | 善恩康生物科技(苏州)有限公司 | 一种复合益生菌及其延缓衰老和抗氧化的作用 |
CN114732834A (zh) * | 2022-03-25 | 2022-07-12 | 善恩康生物科技(苏州)有限公司 | 发酵乳杆菌在制备预防和/或治疗血栓产品中的应用 |
CN114854647A (zh) * | 2022-06-16 | 2022-08-05 | 广西大学 | 发酵乳杆菌及其培养和应用 |
CN116836879A (zh) * | 2023-07-21 | 2023-10-03 | 四川大学 | 一株具有结肠癌细胞生长抑制作用的发酵乳杆菌及其应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008093670A1 (ja) * | 2007-02-01 | 2008-08-07 | Tk Bio-Research Laboratory Co., Ltd. | ブドウ果皮・種子乳酸菌発酵物、並びにそれを利用した医薬及び食品 |
CN109481476A (zh) * | 2018-12-29 | 2019-03-19 | 重庆第二师范学院 | 发酵乳杆菌cqpc04在制备改善溃疡性结肠炎的食品或药品中的应用 |
CN109628346A (zh) * | 2018-12-29 | 2019-04-16 | 重庆第二师范学院 | 发酵乳杆菌cqpc04及其在制备改善便秘的食品中的应用 |
CN110250454A (zh) * | 2019-06-03 | 2019-09-20 | 中国海洋大学 | 一种生产葡萄皮酵素的方法 |
-
2022
- 2022-01-28 CN CN202210107018.6A patent/CN114191490A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008093670A1 (ja) * | 2007-02-01 | 2008-08-07 | Tk Bio-Research Laboratory Co., Ltd. | ブドウ果皮・種子乳酸菌発酵物、並びにそれを利用した医薬及び食品 |
CN109481476A (zh) * | 2018-12-29 | 2019-03-19 | 重庆第二师范学院 | 发酵乳杆菌cqpc04在制备改善溃疡性结肠炎的食品或药品中的应用 |
CN109628346A (zh) * | 2018-12-29 | 2019-04-16 | 重庆第二师范学院 | 发酵乳杆菌cqpc04及其在制备改善便秘的食品中的应用 |
CN110250454A (zh) * | 2019-06-03 | 2019-09-20 | 中国海洋大学 | 一种生产葡萄皮酵素的方法 |
Non-Patent Citations (2)
Title |
---|
JIA LIU,等: "Exploring the Antioxidant Effects and Periodic Regulation of Cancer Cells by Polyphenols Produced by the Fermentation of Grape Skin by Lactobacillus plantarum KFY02", 《BIOMOLECULES》 * |
JIA LIU,等: "Grape skin fermentation by Lactobacillus fermentum CQPC04 has anti-oxidative effects on human embryonic kidney cells and apoptosispromoting effects on human hepatoma cells", 《RSC ADVANCES》 * |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114732834A (zh) * | 2022-03-25 | 2022-07-12 | 善恩康生物科技(苏州)有限公司 | 发酵乳杆菌在制备预防和/或治疗血栓产品中的应用 |
WO2023179274A1 (zh) * | 2022-03-25 | 2023-09-28 | 善恩康生物科技(苏州)有限公司 | 发酵乳杆菌在制备预防和/或治疗血栓产品中的应用 |
CN114642686A (zh) * | 2022-04-29 | 2022-06-21 | 善恩康生物科技(苏州)有限公司 | 一种复合益生菌及其延缓衰老和抗氧化的作用 |
CN114854647A (zh) * | 2022-06-16 | 2022-08-05 | 广西大学 | 发酵乳杆菌及其培养和应用 |
CN114854647B (zh) * | 2022-06-16 | 2023-08-29 | 广西大学 | 发酵乳杆菌及其培养和应用 |
CN116836879A (zh) * | 2023-07-21 | 2023-10-03 | 四川大学 | 一株具有结肠癌细胞生长抑制作用的发酵乳杆菌及其应用 |
CN116836879B (zh) * | 2023-07-21 | 2024-01-23 | 四川大学 | 一株具有结肠癌细胞生长抑制作用的发酵乳杆菌及其应用 |
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