南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (5): 867-875.doi: 10.12122/j.issn.1673-4254.2024.05.08

• 基础研究 • 上一篇    下一篇

副干酪乳杆菌TK1501后生元的制备及对幽门螺旋杆菌的抑制作用

聂金蕊1(), 吴亚慧1, 韩雪梅2, 李亚琪1, 王海宽1(), 张会图1()   

  1. 1.天津科技大学生物工程学院,天津 300457
    2.天津创源生物技术有限公司,天津 300457
  • 收稿日期:2023-11-10 出版日期:2024-05-20 发布日期:2024-06-04
  • 通讯作者: 王海宽,张会图 E-mail:nie15128928292@163.com;hkwang@tust.edu.cn;hzhang@tust.edu.cn
  • 作者简介:聂金蕊,在读硕士研究生,E-mail: nie15128928292@163.com
  • 基金资助:
    国家自然科学基金(82073743)

Preparation of Lactobacillus paracei TK1501 postbiotic and its inhibitory effect against Helicobacter pylori infection in mice

Jinrui NIE1(), Yahui WU1, Xuemei HAN2, Yaqi LI1, Haikuan WANG1(), Huitu ZHANG1()   

  1. 1.School of Biological Engineering, Tianjin University of Science and Technology, Tianjin 300457, China
    2.Tianjin Chuangyuan Biotechnology Co. , Tianjin 300457, China
  • Received:2023-11-10 Online:2024-05-20 Published:2024-06-04
  • Contact: Haikuan WANG, Huitu ZHANG E-mail:nie15128928292@163.com;hkwang@tust.edu.cn;hzhang@tust.edu.cn
  • Supported by:
    National Natural Science Foundation of China(82073743)

摘要:

目的 对副干酪乳杆菌(L. paracasei TK1501)发酵大豆产物中的后生元进行分离提取和体内外抑菌活性进行研究,评估该后生元在治疗幽门螺旋杆菌(Hp)感染的作用。 方法 L. paracasei TK1501在无氧密闭的环境内以37 ℃、固态发酵参数中料水比1∶1.5,接种量5×107 CFU/mL,培养发酵32 h后,通过大孔树脂XAD-16N吸附法、阳离子交换色谱法、高效液相色谱法分离纯化,对L. paracasei TK1501后生元进行稳定性及抗菌效果分析。将50只C57雄性小鼠随机分为空白组、模型组、阴性对照组、低剂量组(0.02 mL/只)和高剂量组(0.1 mL/只),10只/组,连续灌胃4周。取血清观察胃部炎症因子TNF-α、IL-1β表达水平变化,取小鼠胃组织进行HE染色。 结果 L. paracasei TK1501后生元易被蛋白酶降解,热稳定性好,对酸碱、有机溶剂等不利因素有较好的耐受性(P<0.05);在体外试验中,对金黄色葡萄球菌、Hp等均有较好的杀灭作用;在动物实验中,与模型组相比,L. paracasei TK1501后生元高剂量组明显改善Hp感染(P<0.05),胃部炎症因子TNF-α、IL-1β表达水平变化明显下降(P<0.05)。 结论 L. paracasei TK1501后生元对Hp、金黄色葡萄球菌等具有较好的抑制和杀灭效果,而在相同条件下对肠道中的正常菌群则无明显影响。

关键词: 副干酪乳杆菌TK1501, 分离, 抗幽门螺旋杆菌

Abstract:

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori (Hp) infection in mice. Methods L. paracasei TK1501 was cultured for 32 h at 37 ℃ in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL. The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption, cation exchange chromatography and HPLC, and its stability and antibacterial activity were assessed. The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection, which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL. Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments, and gastric tissues of the mice were collected for HE staining. Results L. paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid, base, and organic solvents. In the in vitro experiment, the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus, Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract. In the mouse models, treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice. Conclusion L. paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

Key words: L. paracasei TK1501, isolation, Helicobacter pylori infection