Combined Probiotics and Antioxidants Attenuate LPS-Induced Acute Intestinal Inflammation via Multi-Pathway Regulation in a Preventive Mouse Model.

Abstract

Objectives: This study aimed to explore the combined regulatory effects of probiotics combined with bioactive compounds on intestinal immunity and mucosal barrier function. Methods: Fifty specific-pathogen-free (SPF) male Kunming mice (5-6 weeks old, weighing 20 ± 2 g) were randomly assigned to five groups (n = 10 per group): blank control (C), lipopolysaccharide-induced model (LPS), probiotic (PB), antioxidant (AO), and combined treatment (PB/AO). The C and LPS groups received daily oral gavage of 0.5 mL sterile 0.9% saline. The PB group received a daily gavage of 1 × 10⁸ CFU/mL Pediococcus acidilactici lindner and Lactobacillus plantarum. The AO group was administered a daily gavage of berberine (30 mg/kg BW), wogonin (30 mg/kg BW), and sodium butyrate (200 mg/kg BW). The PB/AO group received a daily gavage combining the probiotic mixture (1 × 10⁸ CFU/mL) with the plant extracts (30 mg/kg BW berberine, 30 mg/kg BW wogonin, and 200 mg/kg BW sodium butyrate). Following a 14-day preventive intervention, all groups except C were intraperitoneally injected with LPS to induce inflammation. Indices of visceral organs, serum antioxidant and immune parameters, ileal histomorphology, gut microbiota composition, and the expression of key barrier- and inflammation-related genes were comprehensively evaluated. Results: LPS exposure increased liver and spleen coefficients. PB/AO treatment significantly reduced the LPS-induced elevation of liver coefficient to a level comparable to that of the C group; however, no significant improvement in spleen coefficient was observed. In terms of antioxidant and immune indices, all intervention groups increased the levels of SOD, GSH-Px, and T-AOC to varying degrees, and decreased the levels of MDA and inflammatory factors (TNF-α, IL-1β, IL-6, and DAO), with the PB/AO group showing the most significant improvement (p < 0.01). Ileal microbiome analysis showed that PB/AO treatment enriched Firmicutes, Lactococcus and Lactobacillus but decreased Proteobacteria, with Clostridia as signature taxa. Mechanistically, PB/AO suppressed the transcription of NF-κB pathway-related genes (TLR4, MyD88 and NF-κB) and upregulated the expression of tight junction proteins (ZO-1, Claudin-1 and Occludin), thereby strengthening the intestinal barrier. Conclusions: In conclusion, co-administration of microbial probiotics and antioxidants achieves optimal protection against LPS-induced intestinal inflammation by jointly driving anti-inflammatory and antioxidative responses, modulating gut microbiota, and reinforcing barrier integrity.