Aflatoxin B1 (AFB1) is a potent mycotoxin that poses significant risks to food safety, public health, and livestock production. Taurochenodeoxycholic acid (TCDCA), a bioactive bile acid, exhibits hepatoprotective and intestinal barrier-modulating potential; however, its role in mitigating AFB1 toxicity remains unclear. This study employed a broiler model to evaluate the protective effects of TCDCA against AFB1-induced hepatic and intestinal injury. The results demonstrated that TCDCA treatment significantly improved growth performance impaired by AFB1 exposure. TCDCA alleviated hepatic lesions, decreased serum hepatic enzyme activities, and suppressed hepatocyte apoptosis and oxidative stress. Mechanistically, TCDCA modulated AFB1 detoxification by downregulating hepatic cytochrome P450 enzymes (CYP1A2 and CYP3A4), thereby reducing the formation of AFB1-DNA adducts. Furthermore, TCDCA attenuated pyroptosis and inflammation by lowering serum inflammatory cytokine contents and inhibiting activation of the hepatic NLRP3/Caspase-1/GSDMD signaling pathway. In the intestine, TCDCA improved the villus morphology, upregulated the expression of tight junction proteins including zonula occludens-1 (ZO-1), claudin-1 (CLDN1), occludin (Occludin), and mucin-2 (MUC2), reduced serum diamine oxidase (DAO) activity and D-lactate level, and restored secretory immunoglobulin A (sIgA) content, thereby strengthening intestinal barrier integrity. Additionally, 16S rRNA sequencing revealed that TCDCA mitigated AFB1-induced gut microbiota dysbiosis by increasing the abundance of Akkermansia and decreasing harmful genera such as Megamonas. Collectively, TCDCA mitigates AFB1-induced growth suppression and hepatotoxicity through mechanisms involving metabolic regulation, antioxidation, inhibition of pyroptosis and inflammation, reinforcement of intestinal barrier function, and modulation of gut microbiota.

Aflatoxin B1 (AFB1) induces hepatocellular damage through its metabolite aflatoxin B1-8,9-epoxide (AFBO), which is produced in endoplasmic reticulum (ER) via cytochrome P450 (CYP450) enzymes. To investigate the effect of ferulic acid (FA) on AFB1-induced broiler liver damage, one-day-old Arbor Acres broilers were exposed to AFB1 (4 mg/kg) and treated with different doses of FA (60 mg/kg, 120 mg/kg, and 240 mg/kg) continuously for 28 days. The production performance, biochemical indicators, morphological changes, CYP450 enzymes’ expression in ER, interactions between small molecules and CYP450 enzymes, and CYP450 enzymes’ protein secondary structure were investigated. The results showed the following: (I) FA promoted broiler growth and reduced AFBO production. (II) AFB1-induced changes in serological indicators (AST, ALT, ALP, γ-GT, TBA, TG) and biochemical parameters (GST, SOD, MDA, ROS), which were reversed by FA. (III) AFB1-induced liver morphological changes and apoptosis were obviously alleviated by FA. (IV) AFB1-induced up-regulation of CYP1A5, CYP2A6, CYP2W1, and CYP3A4 in ER were reduced by FA. (V) The binding affinity of FA to CYP1A5 is lower than that of AFB1 to CYP1A5, and the binding affinity of FA to CYP2W1 is similar to that of AFB1 to CYP2W1. (VI) The contents of α helix, β sheet, β turn, and random coil in chicken CYP1A5 were 59.6%, 7.8%, 13.6%, and 19.0% respectively, and those in chicken CYP2W1 were 32.2%, 17.1%, 16.8%, and 33.9% respectively. In conclusion, FA can promote broiler growth and alleviate AFB1-induced hepatotoxicity via inhibiting and conjugating CYP450 enzymes, thus reducing AFBO formation and oxidative damage.

Background: Metabolic dysfunction-associated steatotic liver disease (MASLD) is now the most prevalent chronic liver disease worldwide, imposing a severe public health burden. Its core pathological hallmark is excessive hepatic lipid accumulation driven by systemic metabolic dysregulation. Concomitant hepatocellular injury impairs hepatic lactate clearance, leading to aberrant lactate buildup in the liver microenvironment. However, the causal role of lactate in exacerbating liver lipid metabolism dysfunction and driving the progression of MASLD remains unclear. Methods: First, we performed a comprehensive bioinformatic analysis of publicly available transcriptomic datasets. Mining of the Gene Expression Omnibus (GEO) database showed that lactate dehydrogenase (LDH) expression was significantly upregulated in liver tissues from both metabolic dysfunction-associated fatty liver disease (MASLD) patients and MASLD mouse models. Next, network pharmacology approaches were employed to predict putative molecular targets that could mediate lactate’s biological effects. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that these candidate targets were predominantly enriched in pathways governing fatty acid metabolism and long-chain fatty acid transport. Molecular docking and molecular dynamics simulations further suggested possible interactions and supported the prioritization of cluster of differentiation 36 (CD36) as candidate lipid metabolism regulators potentially involved in lactate-mediated effects. Finally, liver-specific Ldha knockdown mice (AAV8-TBG-shRNA) and free fatty acid-induced steatotic AML12 hepatocytes were used to investigate the functional relevance of these findings in vivo and in vitro. Results: Network pharmacology analyses preliminarily identified the PPAR signaling pathway as a candidate pathway potentially linking lactate to MASLD. Experimental results showed that exogenous lactate administration was associated with significantly increased lipid accumulation in steatotic AML12 hepatocytes and the livers of MASLD mice, manifested as elevated triglyceride levels and enhanced lipid droplet formation, accompanied by upregulated expression of PPARγ and CD36. Conversely, inhibiting endogenous lactate production or silencing PPARγ or CD36 attenuated this lipid-accumulation phenotype and significantly reduced intracellular triglyceride levels. Conclusions: In conclusion, these findings indicate that lactate exposure is associated with hepatic lipid accumulation and upregulation of the PPARγ/CD36 axis. Pharmacological inhibition or silencing of PPARγ or CD36 attenuates this phenotype, suggesting that this pathway may contribute to lactate-associated hepatic steatosis and potentially accelerate MASLD progression.

Background: Organic acidifiers are promising alternatives to in-feed antibiotics, but the effects of sodium diformate (NaDF) on intestinal health and cecal microbiota in laying hens remain unclear. Objective: This study aimed to investigate the effects of dietary NaDF supplementation on immune organ indices, pro-inflammatory cytokines, intestinal morphology, and cecal microbiota in Hy-Line Brown laying hens. Methods: A total of 720 twenty-one-week-old hens were randomly assigned to 6 groups (6 replicates of 20 hens each). Hens were fed a basal diet supplemented with 0 g/kg, 1 g/kg, 2 g/kg, 3 g/kg, 4 g/kg, or 5 g/kg NaDF for 24 weeks following a 1-week pre-feeding period. Results: Dietary NaDF supplementation showed no significant effect on spleen coefficient or the expression of pro-inflammatory cytokines across intestinal segments (P > 0.05). Notably, 1 g/kg to 4 g/kg NaDF significantly increased duodenal villus height (P < 0.05). The ratio of villus height to crypt depth was not significantly affected, with a linear increasing trend observed (P < 0.05), peaking at 4 g/kg. Ileal villus height was significantly increased (P < 0.05), with linear and quadratic responses (P = 0.028, P = 0.004). Cecal microbiota analysis revealed no significant changes in α diversity or microbial composition at the phylum level (P > 0.05). However, supplementation with 4 g/kg NaDF significantly reduced the abundance of Parabacteroides and Mordavella massiliensis (P < 0.05), while Alistipes and Prevotella showed increasing trends (P = 0.083, P = 0.055). Conclusion: Dietary NaDF supplementation improved duodenal and ileal morphology and exerted minor effects on specific bacterial genera, thereby promoting intestinal health in laying hens. However, it did not significantly affect the spleen index or inflammatory cytokine expression. The 4 g/kg level showed the most consistent improvements in intestinal morphology.