促进GLP-1分泌的益生菌筛选及其安全性评价

doi:10.19586/j.2095-2341.2025.0158

生物技术进展 ›› 2026, Vol. 16 ›› Issue (3): 654-664.DOI: 10.19586/j.2095-2341.2025.0158

促进GLP-1分泌的益生菌筛选及其安全性评价

杜晓娜¹(), 马凤杰¹^,², 冯丰珍², 苏林浩¹^,², 张丽珊², 徐炜²^,³^,⁴, 钱凯³^,⁴, 宋小锋¹()

^1.豫北医学院生命科学技术学院，河南新乡 453003
^2.厦门承葛生物科技有限公司，福建厦门 361000
^3.江西承葛生物科技有限公司，江西宜春 336000
^4.宜春学院医学院，江西宜春 336000

收稿日期:2025-11-10 接受日期:2026-03-30 出版日期:2026-05-25 发布日期:2026-07-14
通讯作者: 宋小锋
作者简介:杜晓娜 E-mail: duxiaona_87@163.com；
基金资助:
河南省生物医药重点学科项目(zdxk2024002);河南省科技发展计划（科技攻关）项目(252102310465);福建省科技计划项目(2024T3073)

Screening and Safety Evaluation of Probiotics Promoting GLP-1 Secretion

Xiaona DU¹(), Fengjie MA¹^,², Fengzhen FENG², Linhao SU¹^,², Lishan ZHANG², Wei XU²^,³^,⁴, Kai QIAN³^,⁴, Xiaofeng SONG¹()

^1.College of Life Sciences and Technology，North Henan Medical University，Henan Xinxiang 453003，China
^2.Xiamen Chengge Biotechnology Corporation Limited，Fujian Xiamen 361000，China
^3.Jiangxi Chengge Biotechnology Co. ，Ltd. ，Jiangxi Yichun 336000，China
^4.School of Medicine，Yichun University，Jiangxi Yichun 336000，China

Received:2025-11-10 Accepted:2026-03-30 Online:2026-05-25 Published:2026-07-14
Contact: Xiaofeng SONG

摘要/Abstract

摘要：

2型糖尿病可引发多种严重并发症，影响全球超4亿人的健康。有研究发现，益生菌可刺激肠道L细胞分泌胰高血糖素样肽-1（glucagon-like peptide-1，GLP-1），继而达到显著的降糖效果。筛选具有显著促进GLP-1分泌的益生菌菌株，旨在为2型糖尿病提供新的治疗策略。通过免疫酶联吸附测定法（enzyme-linked immunosorbent assay，ELISA）对5株候选菌株促进小鼠小肠内分泌细胞（small intestine endocrine cell line，STC-1）分泌GLP-1的含量进行比较，筛选出GLP-1分泌量最高的菌株；评估该菌株对糖相关代谢酶（α-淀粉酶）的抑制活性；通过生理生化实验对菌株进行安全性评价，为其应用提供数据支持；利用荧光定量PCR检测益生菌干预下小鼠小肠内分泌细胞STC-1的G蛋白偶联受体（Gpr41和Gpr43）的相对表达情况，对其降糖机制进行初步探索。结果发现，发酵粘液乳杆菌TH02105（Limosilactobacillus fermentum TH02105）发酵上清液可促进STC-1细胞分泌GLP-1，分泌量为140.340 ng·mL^-1，显著高于其他菌株（P0.001），其对α-淀粉酶的抑制率达66.30%。L. fermentum TH02105对数生长期为2~8 h，最适温度为37 ℃，最适pH为6；该菌在人工模拟肠液、胃液2.5 h后存活率均在50%以上，对人结肠腺癌细胞Caco-2的黏附数量为334.02 CFU·cell^-1；同时，抗生素呋喃妥因、米诺环素、多黏菌素B、环丙沙星、氯霉素、诺氟沙星、左氟沙星均可抑制该菌活性。该菌可显著上调Gpr41/43基因的相对表达量（P0.01）。研究筛选出的L. fermentum TH02105可显著促进STC-1细胞分泌GLP-1，发挥降糖作用；同时该菌对胃肠液有较好耐受性，对Caco-2黏附性较好，有利于发挥定植作用，为研发糖尿病防治药物和保健品提供潜在的菌株材料。

关键词: 2型糖尿病, GLP-1, 发酵粘液乳杆菌, α-淀粉酶, Caco-2

Abstract:

Type 2 diabetes mellitus can lead to a variety of severe complications and affects more than 400 million people worldwide. Studies have found that probiotics can stimulate intestinal L cells to secrete glucagon-like peptide-1 （GLP-1）， thereby achieving a significant blood glucose-lowering effect. This study aimed to screen bacterial strains that significantly promote GLP-1 secretion， with the goal of providing a new therapeutic strategy for type 2 diabetes mellitus. The enzyme-linked immunosorbent assay （ELISA） was used to compare the GLP-1 secretion levels of 5 candidate bacterial strains in stimulating the mouse small intestine endocrine cell line （STC-1）， so as to screen out the strain with the highest GLP-1 secretion. The inhibitory activity of this strain against glucose-related metabolic enzymes （α-amylase） was evaluated. Physiological and biochemical experiments were conducted to assess the safety of the strain， providing data support for its application. Quantitative real-time PCR （qPCR） was used to detect the relative expression levels of G protein-coupled receptors （Gpr41 and Gpr43）， so as to conduct a preliminary exploration of its blood glucose-lowering mechanism. The fermentation supernatant of Limosilactobacillus fermentum TH02105 could promote GLP-1 secretion in STC-1 cells， with a secretion level of 140.340 ng·mL^-1， which was significantly higher than that of other strains （P0.001）. Its inhibition rate against α-amylase reached 66.30%. The logarithmic growth phase of L. fermentum TH02105 was 2~8 h， with an optimal temperature of 37 ℃ and an optimal pH of 6. After being treated with artificial simulated intestinal fluid and gastric fluid for 2.5 h， the survival rate of this strain was more than 50%， and the number of its adhesion to human colonic adenocarcinoma cells Caco-2 was 334.02 CFU·cell^-1. Meanwhile， the antibiotics nitrofurantoin， minocycline， polymyxin B， ciprofloxacin， chloramphenicol， norfloxacin， and levofloxacin could all inhibit the activity of this strain. In addition， this strain significantly upregulated the relative expression levels of Gpr41/43 genes （P0.01）. A strain of L.fermentum TH02105 was screened out， which can significantly promote GLP-1 secretion and exert a blood glucose-lowering effect. At the same time， this strain has good tolerance to gastrointestinal fluids and good adhesion to Caco-2 cells， which is conducive to its colonization. The study provides a potential bacterial strain for the development of drugs and health products for the prevention and treatment of diabetes.

Key words: type 2 diabetes mellitus, GLP-1, Limosilactobacillus fermentum, α-amylase, Caco-2

中图分类号:

Q939.9

杜晓娜, 马凤杰, 冯丰珍, 苏林浩, 张丽珊, 徐炜, 钱凯, 宋小锋. 促进GLP-1分泌的益生菌筛选及其安全性评价[J]. 生物技术进展, 2026, 16(3): 654-664.

Xiaona DU, Fengjie MA, Fengzhen FENG, Linhao SU, Lishan ZHANG, Wei XU, Kai QIAN, Xiaofeng SONG. Screening and Safety Evaluation of Probiotics Promoting GLP-1 Secretion[J]. Current Biotechnology, 2026, 16(3): 654-664.

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链接本文: https://swjsjz.magtechjournal.com/CN/10.19586/j.2095-2341.2025.0158

https://swjsjz.magtechjournal.com/CN/Y2026/V16/I3/654

图/表 9

表1 发酵粘液乳杆菌TH02105对α-淀粉酶抑制活性测定体系

Table 1 Determination system of α-amylase inhibitory activity of L. fermentum TH02105

试剂	样品管	空白管	对照管
PBS缓冲液（0.2 mol·L^-1 pH 7.0）	/	250 µL	250 µL
α-淀粉酶（4 U·mL^-1）	250 µL	/	250 µL
样品（LGG/菌株CFS）	250 µL	250 µL	/
37 ℃孵育10 min（淀粉溶液于37 ℃预热）
可溶性淀粉溶液(1% w/v，质量体积分数)	500 µL	500 µL	500 µL
37 ℃孵育5 min
DNS试剂	1 000 µL	1 000 µL	1 000 µL
沸水浴5 min，迅速冷却至室温后加13 mL蒸馏水稀释，再稀释10倍，540 nm处测定吸光度

图1 不同处理对GLP-1分泌量的影响A：6株菌株对GLP-1分泌量的影响；B：灭活L. fermentum TH02105和灭活LGG对GLP-1的分泌量的影响；C：L. fermentum TH02105和LGG活菌对GLP-1的分泌量的影响；D：L. fermentum TH02105和LGG上清以及无细胞提取物对GLP-1的分泌量的影响(n=3）。 *、**、****分别表示差异在P0.05、P0.01、P0.000 01水平上具有统计学意义，ns表示无统计学意义。

Fig. 1 Effects of different treatments on GLP-1 secretion volume

图2 L. fermentum TH02105菌液浓度对STC-1细胞活性的影响（n=6）

Fig. 2 The effect of L. fermentum TH02105 concentration on the activity of STC-1 cells (n=6)

图3 L. fermentum TH02105对α-淀粉酶的抑制率（n=3）

Fig. 3 The inhibition rate of L. fermentum TH02105 on α-amylase (n=3)

图4 L. fermentum TH02105的生长曲线、最适pH和最适温度

Fig. 4 Determination of growth curve, optimal pH and optimal temperature of L. fermentum TH02105

表2 L. fermentum TH02105抗生素敏感性结果

Table 2 Antibiotic susceptibility of L. fermentum TH02105

抗生素/每片药物含量	抑菌圈直径评判标准/mm	抑菌圈直径/mm	敏感性
大观霉素/100 μg	R≤14， 15I17， S≥18	14.45±0.47	R
多黏菌素B/30 μg	R≤8， 9I11， S≥12	16.85±0.15	S
呋喃妥因/300 μg	R≤14， 15I16， S≥17	15.8±1.82	I
环丙沙星/5 μg	R≤15， 16I20， S≥21	23.9±0.76	S
氯霉素/30 μg	R≤12， 13I17， S≥18	20.35±0.45	S
米诺环素/30 μg	R≤14， 15I18， S≥19	17.6±0.25	I
诺氟沙星/10 μg	R≤12， 13I16， S≥17	19.9±1.92	S
左氧氟沙星/5 μg	R≤13， 14I16， S≥17	23.4±1.42	S

图5 L. fermentum TH02105对人结直肠腺癌细胞Caco-2的黏附图A：空白细胞对照；B：发酵乳杆菌TH02105黏附后的细胞图

Fig. 5 Adhesion of L. fermentum TH02105 to Caco-2 cells

图6 L. fermentum TH02105抗生素敏感性

Fig. 6 Antibiotic sensitivity of L. fermentum TH02105 fermentation

图7 Gpr41/43在STC-1细胞中的相对表达量（n=3）注：**表示差异在P0.01水平上具有统计学意义。

Fig. 7 Relative expression levels of Gpr41/43 in STC-1 cells (n=3)

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促进GLP-1分泌的益生菌筛选及其安全性评价

Screening and Safety Evaluation of Probiotics Promoting GLP-1 Secretion

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