Lactiplantibacillus plantarum JY039 Ameliorates Necrotizing Enterocolitis via Intestinal Stem Cell Regeneration.

L. plantarum JY039 was selected from 25 probiotic strains based on superior 2'-FL metabolism, intestinal adhesion, and Lgr5 induction in TNF-α-damaged organoids.

• Selection was performed from a pool of 25 strains with statistical significance at p < 0.05.
• Selection criteria included 2'-fucosyllactose (2'-FL) utilization efficiency, intestinal adhesion capacity, and ability to induce Lgr5 expression in TNF-α-damaged intestinal organoids.
• JY039 demonstrated versatile carbon metabolism properties.

JY039 elevated intestinal stem cell markers Olfm4 and Ascl2 approximately 4-fold in TNF-α-injured organoids compared to NEC models.

• ISC markers Olfm4 and Ascl2 were elevated approximately 4-fold versus NEC models.
• Proliferation markers c-Myc, Ki67, and cyclins were elevated approximately 3-fold versus NEC models.
• EdU intensity, a measure of cellular proliferation, increased 4-fold versus NEC models.
• These effects were observed in TNF-α-damaged intestinal organoids treated with JY039.

JY039 demonstrated clinical safety characteristics based on antibiotic sensitivity profiling and carbon metabolism analysis.

• JY039 showed sensitivity to 11 antibiotics, indicating susceptibility to standard clinical treatments.
• The strain demonstrated versatile carbon metabolism.
• These characteristics were assessed as indicators of clinical safety for potential therapeutic use.

In NEC rats, JY039 administered at 10^9 CFU/mL reduced pathological scores and intestinal permeability.

• JY039 was administered at a dose of 10^9 CFU/mL in the NEC rat model.
• Treatment reduced pathology scores in NEC rats.
• Intestinal permeability was reduced following JY039 treatment.
• Tight junction proteins were restored in the intestinal epithelium of treated rats.

JY039 modulated inflammatory cytokine profiles in NEC rats, decreasing pro-inflammatory markers and increasing anti-inflammatory IL-10.

• TNF-α levels were decreased following JY039 treatment (p < 0.05).
• IL-6 levels were decreased following JY039 treatment (p < 0.05).
• Myeloperoxidase (MPO) activity was decreased following JY039 treatment (p < 0.05).
• IL-10, an anti-inflammatory cytokine, was increased following JY039 treatment (p < 0.05).

JY039 treatment enriched beneficial gut microbiota, specifically Lactococcus and Akkermansia, in NEC rats.

• Akkermansia, a genus associated with gut barrier integrity and health, was enriched following JY039 treatment.
• Lactococcus abundance was increased in JY039-treated NEC rats.
• JY039 treatment improved the microbial imbalance (dysbiosis) characteristic of NEC.
• Microbial modulation occurred alongside activation of intestinal stem cell regeneration.

NEC is driven by dysbiosis-induced inflammation and barrier damage and represents a serious condition in premature infants.

• NEC pathogenesis involves intestinal dysbiosis leading to inflammation and barrier damage.
• The condition primarily affects premature infants.
• The study identified intestinal stem cell (ISC) regeneration as a key therapeutic target for NEC intervention.
• 2'-fucosyllactose (2'-FL), a human milk oligosaccharide, was used as a relevant substrate for probiotic screening in the context of infant nutrition.