Garlic diallyl disulfide and diallyl trisulfide mitigates effects of pro-oxidant induced cellular stress and has immune modulatory function in LPS-stimulated porcine epithelial cells.

ABSTRACT

The objective of the current study was to determine if garlic-derived diallyl disulfide (DADS) and diallyl trisulfide (DATS) could mitigate oxidative and endotoxin stress, using an intestinal porcine epithelial cell (IPEC-J2) model. The experiment was arranged as a 2 × 2 × 2 factorial of DADS + DATS (0 or 18 µM), pro-oxidant stressor (hydrogen peroxide at 0 or 100 µM), and endotoxin stressor (lipopolysaccharide [LPS] at 0 or 10 µg/mL) with 8 replicates per treatment. Cells were incubated with DADS + DATS for 18 h, LPS for 6 h, then with hydrogen peroxide for 3 h. Gene expression was measured by RT-PCR for cytokines, interleukin 8 (IL-8) and tumor necrosis factor α (TNF-α), and tight junction proteins, claudin 1 (CL-1), occludin (OC), and zonula occludens 1 (ZO-1). Trans-epithelial electrical resistance (TEER), the antioxidant enzyme catalase, and apical secretion of IL-8 protein into the incubation medium was also measured. There was an increase ( < 0.01) in TNF-α and IL-8 gene expression due to LPS, although there was no effect of hydrogen peroxide or DADS + DATS. Furthermore, there was a tendency for an increase ( = 0.08) in ZO-1 gene expression due to DADS + DATS. Treatment with DADS + DATS and hydrogen peroxide did not affect TEER, although there was a decrease ( = 0.02) in TEER with LPS incubation. Treatment of cells with hydrogen peroxide reduced catalase activity ( < 0.01), which was restored with pre-incubation of DADS + DATS ( < 0.10). There was an increase ( < 0.01) in IL-8 secretion due to LPS, which was further augmented ( < 0.01) by pre-incubation with DADS + DATS. Based on the results from the current study, DADS + DATS can ameliorate oxidative effects of hydrogen peroxide, as well as alter IL-8 secretion in LPS-treated IPEC-J2 cells.