Characterizing biological functions of Lactobacillus brevis 1E1 in porcine jejunal epithelial cell line (IPEC-J2)

• Published in: Journal of animal science Vol. 96; pp. 68 - 69
• Main Authors: Wang, X, Tsai, T, Chewning, J, Maxwell, C
• Format: Journal Article
• Language: English
• Published: Champaign Oxford University Press 01.12.2018
• Subjects: Apoptosis; Bcl-2 protein; Bcl-3 protein; Data processing; Dextran; Dietary supplements; E coli; Epithelial cells; FADD protein; Fluorescein; Fluorescein isothiocyanate; Gene expression; Hogs; Hydrogen peroxide; Inflammation; Integrity; Interleukin 6; Interleukin 8; Lactobacillus brevis; Lipopolysaccharides; Permeability; Spectrometry; Suidae; Swine; TLR4 protein; Toll-like receptors; Tumor necrosis factor-α; Variance analysis; Weaning
• ISSN: 0021-8812; 1525-3163

Our previous finding suggested that supplementing Lactobacillus brevis 1E1 (L. 1E1) to pre-weaning piglets resulted in heavier nursery pigs. This project aimed at further understanding the biological functions of L. 1E1. Confluent porcine jejunal epithelial (IPEC-J2) cells were either treated with: a) Placebo, b) 1 pg/mL of Escherichia coli (0111:B4-derived lipopolysaccharide (LPS), c) heat-killed L. 1E1 (107 cells/ml, HK1E1), d) live L. 1E1 (107cells/ml, Live1E1), e) L. 1E1 + LPS, f) HK1E1 + LPS for 4 hours to evaluate the pro- or anti-inflammatory responses and cellular apoptosis through gene expressions of IL-6, IL-8, TNF-a, TLR4, A20, Bcl-3, FADD, Fas, and Bcl-2 or pre-cultured with or without 107 CFU/ml L. 1E1 for 4 hours and challenged with 0 or 10 mM of H2O2 for two hours in transwell plate to assess barrier integrity by measuring transepithelial electrical resistance (TEER), and permeability (Fluorescein isothiocyanate-FITC dextran 70 kDa) using Spectrometry. Data were analyzed using the SAS GLM procedures with treatments as main effect in ANOVA. Unlike HK1E1 with few or no difference in gene expressions, co-culturing IPEC-J2 with Live L. 1E1 upregulated TNF-a, A20 (anti-inflammatory cytokine), Bcl-2 (anti-apoptosis gene), while suppressed IL-6, Bcl3, FADD (apoptosis initiator), and Fas when compared to Placebo. Moreover, Live1E1 mitigated IL-6 but promoted TNF-a, A20, and Bcl-2, while HK1E1 increased A20, Bcl-2, and FADD gene expressions when co-challenged with LPS. Adding H2O2 significantly impaired the monolayer integrity as indicated by a 60% reduction on TEER value (5625 vs. 1925; P < 0.01) and increased the permeability (6.6 vs. 9; P < 0.01). Preincubation with L. 1E1 or HK1E1 failed to counteract the barrier damage caused by H2O2. In conclusion, the growth promoting effect of L. 1E1 observed in nursery pigs may be due to its inflammation-regulatory ability, and L. 1E1 does not have protective effect against hydrogen peroxide.