CN114164149B - 一株缓解肥胖和铅毒性的植物乳杆菌p101及其应用 - Google Patents
一株缓解肥胖和铅毒性的植物乳杆菌p101及其应用 Download PDFInfo
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Abstract
本发明公开了一株缓解肥胖和铅毒性的植物乳杆菌P101及其应用,属于微生物技术领域。本发明植物乳杆菌P101分离自江西省吉安市农家自制酸菜,保藏编号为CCTCC M 2021108,保藏地点为武汉市中国典型培养物保藏中心。本发明植物乳杆菌P101为革兰氏阳性菌,其具有较高的抗氧化活性以及体内外铅吸附特性,在一定程度上能缓解肥胖和铅暴露带来的肝脏氧化损伤。本发明植物乳杆菌P101菌株具有较好的安全和益生性能,该菌株在缓解高糖高脂饮食造成的肥胖及缓解铅暴露毒性的应用中发挥有益作用。
Description
技术领域
本发明属于微生物技术领域,具体涉及一株缓解肥胖和铅毒性的植物乳杆菌P101及其应用。
背景技术
随着人们生活水平的日益提升,肥胖症已成为世界范围内关注的公共卫生问题,其病理基础包括全身性低度慢性炎症及脂质代谢紊乱,而其成因包括久坐、不运动及高糖分高脂肪饮食等不健康的生活方式习惯,使用饮食诱导肥胖已成为肥胖症研究中最常用的方法。研究表明益生菌能调节肠道微生物稳态,并影响包括短链脂肪酸在内的多种关键分子的生物合成及代谢,进而对以脂肪堆积为主要表型的肥胖症起一定的治疗效果。
铅是生活中较常见的重金属污染物质,进入人体的主要途径包括汽车尾气、街面灰尘、不合格的食品包装以及食物链传递。长期处在铅暴露的环境下会诱发神经***及心血管***紊乱、炎症反应、肝脏损伤等,对于青少年还会引起智力障碍。研究表明益生菌一方面能缓解铅引起的氧化应激损伤,另一方面能在人体肠道中吸附铅离子,促使铅随着粪便排出体外。因此,采用益生菌来治疗铅暴露引起的生物毒性成为可能。
植物乳杆菌是一类革兰氏阳性菌,其多为兼性厌氧菌,是公认的益生菌菌种之一。植物乳杆菌与人类生活息息相关,其广泛存在于自然界中,尤其是各类发酵食品中,如蔬菜、肉及乳制品等。同时植物乳杆菌作为益生菌也存在于人体胃肠道中,调节肠道微生物平衡,控制人体体内毒素水平,增强机体免疫力,清除有害物质,对机体健康有重大意义。
发明内容
本发明的目的在于提供一株缓解肥胖和铅毒性的植物乳杆菌P101(Lactobacillus Plantarum P101)及其应用;本发明植物乳杆菌P101具有良好的益生性能,其具备较高的抗氧化活性以及体内外铅吸附特性。
本发明涉及一株缓解肥胖和铅毒性的植物乳杆菌P101,该植物乳杆菌P101于2021年1月19日保存于武汉市中国典型培养物保藏中心(武汉大学),保藏编号为:CCTCC M2021108。
进一步地,所述植物乳杆菌P101分离自农家自制酸菜。
本发明植物乳杆菌P101可在MRS琼脂平板培养基上生长,为革兰氏阳性杆菌,兼性厌氧,不产芽孢。为维持本发明植物乳杆菌的优良性能,将其保存在含25%甘油的MRS混合保存液中,于-80℃冻存。
本发明另一个目的在于提供一株植物乳杆菌P101在制备用于缓解肥胖的药品、食品和保健品中的应用。
进一步地,所述肥胖是由高糖高脂饮食诱导的肥胖症。
进一步地,所述应用中植物乳杆菌P101使用的活菌数为108~109CFU/mL。
本发明另一个目的在于提供一株植物乳杆菌P101在制备用于缓解铅毒性的药品、食品和保健品中的应用。
进一步地,所述铅毒性是低浓度铅暴露引起的生物毒性。
进一步地,所述应用中植物乳杆菌P101使用的活菌数为108~109CFU/mL。
本发明以传统酸菜中分离鉴定的乳杆菌为研究对象,通过动物实验对肥胖症以及铅污染模型进行干预治疗,旨在为益生菌的应用及常见公共卫生问题的治疗提供新思路。
高糖高脂饲料诱导肥胖模型、醋酸铅溶液诱导慢性铅暴露模型小鼠结果显示:植物乳杆菌P101可以有效调节高糖高脂饮食诱导的脂质代谢紊乱,显著地增加小鼠粪便中铅排出并减少脏器中的铅蓄积。本发明植物乳杆菌P101作为益生菌,是一株性能优良且安全性较高的菌种,并能有效缓解饮食诱导的肥胖症及铅暴露引起的生物毒性,因此具有较高的研究价值和应用。
与现有技术相比,本发明的有益效果是:
本发明植物乳杆菌P101对高糖高脂饮食诱导的肥胖症有显著的治疗效果,并能缓解低浓度铅暴露引起的肝脏损伤,本发明扩大了植物乳杆菌P101应用领域,提高了其应用价值。
附图说明
图1是植物乳杆菌P101缓解饮食诱导的肥胖实验小鼠体重增量(不同小写字母之间表示存在显著差异,p<0.05)。
图2是植物乳杆菌P101缓解饮食诱导的肥胖实验小鼠皮下及附睾脂肪系数(不同小写字母之间表示存在显著差异,p<0.05)。
图3是植物乳杆菌P101缓解饮食诱导的肥胖实验小鼠肝脏中TC(总胆固醇)、TG(总甘油三酯)水平(不同小写字母之间表示存在显著差异,p<0.05)。
图4是植物乳杆菌P101缓解饮食诱导的肥胖实验小鼠血清中TC、TG水平(不同小写字母之间表示存在显著差异,p<0.05)。
图5是植物乳杆菌P101缓解饮食诱导的肥胖实验小鼠皮下脂肪组织病理切片分析(400倍)。
图6是植物乳杆菌P101缓解铅毒性实验小鼠粪便、肝脏及肠道铅含量测定。
图7是植物乳杆菌P101缓解铅毒性实验小鼠肝脏组织病理切片分析(400倍)。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚,下面将结合实施例对本发明的技术方案进行清楚、完整地描述。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
实施例1:缓解高糖高脂饮食诱导的肥胖
21只6周龄昆明小鼠适应1周后,随机分为三组:对照组、高糖高脂饮食组和益生菌治疗组,对照组每天喂食日常维持饲料,高糖高脂饮食组和益生菌治疗组喂食高糖高脂饲料,所有饲料及饮用水均自由摄取,持续8周。在第7、8周给对照组与高糖高脂饮食组每天灌胃200μL的磷酸盐缓冲液作为对照,益生菌治疗组每天灌胃200μL(108CFU/mL)的植物乳杆菌P101菌悬液。实验期间每天记录小鼠体重。小鼠在末次给药24h后安乐死,眼部静脉取血后断颈,收集肝脏、皮下脂肪和附睾脂肪用于后续理化分析。肝脏及皮下脂肪组织用石蜡包埋后切成切成5μm的薄片,使用苏木精和伊红染料进行染色,显微镜下观察组织病理形态。
结果如图1所示,高糖高脂饮食能显著增加小鼠体重,而植物乳杆菌P101的干预显著降低小鼠体重的增加。
结果如图2所示,高糖高脂饮食显著增加了小鼠的皮下及附睾脂肪重量,植物乳杆菌P101能将其恢复至正常水平。
结果如图3所示,高糖高脂饮食显著增加了小鼠肝脏中的TC、TG水平,植物乳杆菌P101治疗组下降至正常水平。
结果如图4所示,高糖高脂饮食显著增加了小鼠血清中的TC、TG水平,植物乳杆菌P101治疗组下降至正常水平。
结果如图5所示,高糖高脂饮食组的皮下脂肪细胞尺寸显著增加,植物乳杆菌P101治疗组的皮下脂肪细胞尺寸下降至正常水平。
以上结果表明,植物乳杆菌P101能在一定程度上缓解高糖高脂饮食引起的肥胖症状,包括降低额外能量摄入导致的体重增加,以及血清和肝脏中的总胆固醇和甘油三酯含量,通过对脂肪组织的切片观察也能看出植物乳杆菌对脂肪堆积具有一定缓解作用。
实施例2:缓解铅暴露引起的生物毒性
14只6周龄昆明小鼠适应1周后,随机分为两组:铅暴露组和益生菌治疗组,铅暴露组自由饮用1g/L Pb2+的醋酸铅溶液,益生菌治疗组在饮用1g/L Pb2+的醋酸铅溶液的同时每天灌胃200μL(108CFU/mL)的P101菌悬液,持续2周。小鼠在末次给药24h后安乐死,眼部静脉取血后断颈,收集肝脏、结肠及粪便样本用于后续理化分析。
如图6所示,植物乳杆菌P101治疗组的粪便中铅含量显著上升,同时肝脏及结肠中铅元素含量显著下降,表明植物乳杆菌的干预促进了粪便中的铅***,并因此减少脏器中的铅蓄积。
结果如图7所示,铅暴露组的小鼠肝脏组织切片能明显观察到细胞空泡化以及核固缩、核溶解等坏死等病理症状,而植物乳杆菌P101干预治疗后能缓解铅暴露引起的肝脏细胞空泡化、坏死及炎症浸润,表明植物乳杆菌P101对铅暴露诱导的生物毒性具有显著的治疗效果。
以上结果表明,植物乳杆菌P101能通过促进铅***减少脏器中的铅蓄积,进而缓解铅暴露引起的包括肝脏损伤等生物毒性。
最后应当说明的是:以上实施例仅用以说明本申请的技术方案而非对其限制;尽管参照较佳实施例对本申请进行了详细的说明,所属领域的普通技术人员应当理解:依然可以对本申请的具体实施方式进行修改或者对部分技术特征进行等同替换,其均应涵盖在本申请请求保护的技术方案范围当中。
Claims (2)
1.一株植物乳杆菌P101在制备缓解高糖高脂饮食诱导的肥胖症的药品中的应用;
所述植物乳杆菌P101于2021年1月19日保存于武汉市中国典型培养物保藏中心,保藏编号为:CCTCC M 2021108。
2.如权利要求1所述的植物乳杆菌P101在制备用于缓解低浓度铅暴露引起的肝脏损伤的药品中的应用。
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