Lactobacillus plantarum ZG03 alleviates oxidative stress via its metabolites short-chain fatty acids

doi:10.12122/j.issn.1673-4254.2025.10.18

Abstract

Abstract:

Objective To investigate the efficacy of Lactobacillus plantarum ZG03 (L. plantarum ZG03) for ameliorating oxidative stress in zebrafish. Methods We evaluated the growth pattern of L. plantarum ZG03, observed its morphology using field emission scanning electron microscopy, and assessed its safety and potential efficacy with whole-genome sequencing for genetic analysis. FITC-labeled ZG03 was used to observe its intestinal colonization in zebrafish. In a zebrafish model of 2% glucose-induced oxidative stress, the effect of ZG03 was evaluated by assessing the changes in neutrophils in the caudal hematopoietic tissue (CHT), superoxide dismutase (SOD) activity, reactive oxygen species (ROS) levels, and malondialdehyde (MDA) content. Liquid chromatography-mass spectrometry-based targeted metabolomics was used for analyzing short-chain fatty acids (SCFAs) in the zebrafish, and the antioxidant effects of the key metabolites (acetate, propionate, and caproate) were tested. Results On MRS agar, L. plantarum ZG03 formed circular, smooth, moist, and milky-white colonies with a rod-shaped cell morphology. Genomic analysis revealed abundant sugar metabolism gene clusters. After inoculation of FITC-labeled L. plantarum ZG03 in zebrafish, green fluorescence was clearly observed in the intestinal bulb, mid-intestine, and hind intestine. In zebrafish with glucose-induced oxidative stress, L. plantarum ZG03 significantly reduced ROS levels and the number of neutrophils in the CHT with increased SOD activity. L.plantarum ZG03 significantly increased the content of SCFAs including acetic acid, propionic acid, and caproic acid in zebrafish metabolites. In addition, sodium acetate, sodium propionate, and sodium caproate in the SCFAs significantly increased SOD activity in the zebrafish models. Conclusion L. plantarum ZG03 ameliorates oxidative stress in a glucose-induced zebrafish model through its metabolites, particularly the SCFAs including acetic acid, propionic acid and caproic acid.

Key words: Lactobacillus plantarum ZG03, short-chain fatty acids, glucose, oxidative stress

Shuxian LIN, Lina GUO, Yan MA, Yao XIONG, Yingxi HE, Xinzhu XU, Wen SHENG, Suhua XU, Feng QIU. Lactobacillus plantarum ZG03 alleviates oxidative stress via its metabolites short-chain fatty acids[J]. Journal of Southern Medical University, 2025, 45(10): 2223-2230.

Figures/Tables 8

Fig.1 Growth characteristics of L. plantarum ZG03. A: Growth curve of ZG03. B: Colony morphology of ZG03. C: Field emission scanning electron microscopy (FESEM) images of ZG03.

Fig.2 Genomic characteristics of L. plantarum ZG03. A: Genomic circular map (from outer to inner circles): The first circle represents genomic position information; the second circle shows GC content; the third circle indicates protein-coding genes on the positive strand (marked in red); the fourth circle denotes protein-coding genes on the negative strand (marked in green); the fifth circle displays non-coding elements on the positive strand (marked in blue); the sixth circle represents non-coding elements on the negative strand (marked in purple); and the seventh circle shows long fragment repeat sequences within the genome (marked in orange). B: KEGG enrichment analysis. C: COG enrichment analysis.

Tab.1 Genomic information of L. plantarum ZG03

Feature	Value
Size (bp)	3 613 757
G+C content (%)	46.42
Coding region (%)	80.70
Total genes	3 354
RNA genes	123
rRNA genes	16
tRNA genes	70
Protein-coding genes	3 231
Protein coding genes with enzymes	1 099
Genes with signal petides	176
Genes with transmembrane helices	911

Fig.3 Colonization by L. plantarum ZG03 in the intestinal tract of zebrafish. ***P<0.001 vs control group.

Fig.4 Number of neutrophils in the caudal hematopoietic tissue of zebrafish (n=10). A: Representative images of neutrophils (dashed box) in zebrafish. B: Quantitative analysis of neutrophil numbers in zebrafish. ###P<0.001 vs control group; ***P<0.001 vs model group.

Fig.5 Effect of L. plantarum ZG03 on glucose-induced oxidative stress in zebrafish. A: Representative images of ROS staining in zebrafish. B: Statistical analysis of relative ROS levels (n=6). C: SOD activity in zebrafish (n=3). D: MDA levels in zebrafish (n=3). ##P<0.01 vs control group; *P<0.05, **P<0.01, ***P<0.001 vs model group.

Fig.6 Content of short-chain fatty acid metabolites in zebrafish (n=3). A: Acetic acid content in zebrafish. B: Propionic acid content in zebrafish. C: Hexanoic acid content in zebrafish. ##P<0.01, ###P<0.001vs control group; *P<0.05 vs model group.

Fig.7 Statistical chart of superoxide dismutase (SOD) activity in zebrafish (n=3). ##P<0.001 vs control group,***P<0.001 vs model group.

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