Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2

Abstract

The in vitro anti-inflammatory effect of apigenin and its trimethylated analogue (apigenin-trimethylether) has been investigated in order to evaluate whether these flavonoids could attenuate LPS-induced inflammation in IPEC-J2 non-transformed intestinal epithelial cells. Levels of IL-6, IL-8, TNF-α, and COX-2 mRNA were measured as a marker of inflammatory response. The extracellular H2O2 level in IPEC-J2 cells was also monitored by Amplex Red assay. Our data revealed that both compounds had significant lowering effect on the inflammatory response. Apigenin (at 25 μM) significantly decreased gene expression of IL-6 in LPS-treated cells, while apigenin-trimethylether in the same concentration did not influence IL-6 mRNA level. Both apigenin and apigenin-trimethylether reduced IL-8 gene expression significantly. TNF-α mRNA level was decreased by apigenin-trimethylether, which was not influenced by apigenin. Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one. Furthermore, both flavonoids reduced significantly the level of extracellular H2O2 compared to the control cells. In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.

Chlorogenic Acid Combined with Lactobacillus plantarum 2142 Reduced LPS-Induced Intestinal Inflammation and Oxidative Stress in IPEC-J2 Cells

“…It was found, that level of extracellular H 2 O 2 detected by Amplex Red method was not affected by 10 μg/ml LPS treatment. LPS in higher concentration (50 μg/ml) increased intracellular H 2 O 2 level significantly, however, IPEC-J2 cells are irreversible damaged [19]. Still, it is important to examine the impact of chlorogenic acid on IPEC-J2 cells, because of its possible pro-oxidant effect.…”

“…LPS solutions were prepared freshly prior to each experiment. LPS was added in plain medium at 10 μg/ml on the apical side of the IPEC-J2 layer [19]. After 1 h incubation with LPS and test compounds, cells were washed with plain medium and cultured for additional 1 h for PCR studies.…”

“…The cDNA was diluted 5-fold, before equal amounts were added to duplicate qRT-PCR reactions. Tested genes of interest were IL-8 [22], IL-6 [23], TNF-α [24] and COX-2 [19]. Hypoxanthine phosphoribosyltransferase (HPRT) [25] and Cyclophilin-A (CycA) [24] were used as reference genes.…”

Deoxynivalenol, but not E. coli lipopolysaccharide, changes the response pattern of intestinal porcine epithelial cells (IPEC-J2) according to its route of application

“…Although LPS-dose and -type were comparable to our study, there were differences in IPEC-J2 culture conditions -impermeable supports vs. permeable supports in our study -and possibly in cell culture passages as well as a shorter incubation time for LPS that might have contributed to the apparent differences. Others used a short-term Salmonella-LPS challenge (1 h) in an IPEC-J2 model grown on inserts and reported an upregulation of IL-8 transcripts (Farkas et al, 2015) and a slight increase in IL-8 protein production to the basolateral site (Farkas et al, 2014). In these studies IPEC-J2 were cultured in the presence of antibiotics and challenged with a different type of LPS.…”

Crosstalk Between Nuclear Glucose-Regulated Protein 78 and Tumor Protein 53 Contributes to the Lipopolysaccharide Aggravated Apoptosis of Endoplasmic Reticulum Stress-Responsive Porcine Intestinal Epithelial Cells

“…Treatment with LPS results in the activation of nuclear factor (NF)-κB and subsequent upregulation of proinflammatory cytokines including interleukin (IL)-1α, IL-1β, and IL-6 [19]. Previous studies using the IPEC-J2 cell line have established a model of inflammation using Salmonella enterica-derived LPS [18,20]. However, there are few reports on inflammatory models based on E. coli-derived LPS in intestinal epithelial cells.…”