Lactiplantibacillus plantarum Lp05 protects against ethanol-induced liver injury in zebrafish through metabolic and microbiota modulation

Abstract

This study investigates the protective effects of Lactiplantibacillus plantarum Lp05 on ethanol-induced liver injury in zebrafish (Danio rerio), focusing on its mechanisms related to metabolic pathway regulation and gut microbiota composition. We administered Lp05 to zebrafish exposed to ethanol to assess its impact on liver enzymes and overall liver health. Key metrics included the expression levels of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and changes in gut microbiota composition through 16S rRNA sequencing. Metabolomic analyses were conducted to identify affected metabolic pathways. Treatment with Lp05 significantly increased ADH and ALDH expression, enhancing alcohol metabolism and reducing ALT and AST activities, thereby mitigating liver cell damage. Metabolomic analysis revealed significant modulation of biotin and glycerolipid metabolism pathways, crucial for reducing liver injury. Lp05 also altered gut microbiota, increasing beneficial bacteria such as Fluviicola and Delftia, while decreasing pathogenic bacteria like Acinetobacter and Aeromonas. This bacterial modulation contributed to phenylalanine metabolism regulation, which alleviated intestinal inflammation and liver injury. Additionally, Lp05 promoted the degradation of polycyclic aromatic hydrocarbons (PAHs), reducing their health risks. Lp05 exhibits potential therapeutic effects against ethanol-induced liver injury in zebrafish by enhancing alcohol metabolism, modulating metabolic pathways, and altering gut microbiota. These findings suggest that Lp05 may offer a novel preventative and therapeutic strategy for managing alcoholic liver injury. Alcohol-induced liver injury is a significant global health concern with limited effective therapeutic options. This study highlights the potential of Lactiplantibacillus plantarum Lp05 as a novel probiotic intervention for mitigating ethanol-induced liver damage. By enhancing alcohol metabolism, regulating critical metabolic pathways such as biotin and glycerolipid metabolism, and modulating gut microbiota composition, Lp05 addresses both the metabolic and microbiological aspects of liver health. The promotion of beneficial bacteria and the suppression of pathogenic strains further contribute to alleviating liver injury and systemic inflammation. These findings underscore the therapeutic promise of Lp05 in managing alcoholic liver injury and provide a foundation for its development as a preventative and therapeutic strategy in human applications.

Keywords: Lactiplantibacillus plantarum; 16S rRNA sequencing; Ethanol-induced liver injury; Metabolic pathways; Zebrafish model.

Fig. 1

Zebrafish liver and yolk sac area. The L and yellow dashed lines represent the liver; the Y and red dashed lines represent the yolk sac. Ctl, the control group; M, model group; Pos, positive control group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 2

Zebrafish liver and yolk sac areas. (a) zebrafish liver area; (b) zebrafish yolk sac area. * p value < 0.05; ** p value < 0.01; *** p value < 0.001. Ctl, the control group; M, model group; Pos, positive control group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 3

ADH, ALDH, AST and ALT levels in zebrafish. (a) ADH; (b) ALDH; (c) AST; (d) ALT. * p value < 0.05; ** p value < 0.01; *** p value < 0.001. Ctl, the control group; M, model group; Pos, positive control group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 4

Hiopathological section of zebrafish liver. Red arrows in the figure represent enlarged hepatocyte nucleus, and green arrows represent fatty vacuolar-like degeneration. Ctl, the control group; M, model group; Pos, positive control group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 5

Zebrafish liver ultragraph. (N) nucleus; (M) mitochondria; (RER) rough endoplasmic reticulum; (GL) glycogen; (Mi) microbodies; (ASS) autophagosomes; (LD) lipid droplets. Ctl, the control group; M, model group; Pos, positive control group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 6

Zebrafish Metabolic Figures. (a) PCA plot; (b) volcano plot of differential metabolites for Ctl and M; (c) volcano plot of differential metabolites for Lp05 and M; (d) venn diagram of differential metabolites between Ctl vs. M and Lp05 vs. M; (e) heatmap of the top 30 most differential metabolites of 69 differential metabolites. Ctl, the control group; M, model group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 7

KEGG Pathway Analysis of Differential Metabolites. (a) pathway map of differential metabolites for Ctl vs. M; (b) pathway map of differential metabolites for Lp05 vs. M. The size of the circles in the graph represents the number of differential metabolites contained in that pathway; the larger the circle, the more differential metabolites it contains. The bluer the circle color, more significant the pathway. Ctl, the control group; M, model group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 8

The 16S rRNA sequencing results. (a) species accumulation curve; (b) ACE; (c) Chao1; (d) Shannon; (e) Simpson; (f) LEfSe bae chart; (g) LEfSe cladogram; (h) Pcoa of Beta analysis; (i) heatmap of top 15 significant abundance genus; (j) circos plot of the five most abundant genus. Ctl, the control group; M, model group; Lp05, Lactiplantibacillus plantarum Lp05.

Fig. 9

Stamp analysis of Ctl vs. M and Lp05 vs. M. (a) stamp result of Ctl vs. M; (b) stamp result of Lp05 vs. M. Ctl, control group; M, model group; Lp05, Lactiplantibacillus plantarum Lp05.