Effects of Lactobacillus plantarum 2142 and sodium n-butyrate in lipopolysaccharide-triggered inflammation: Comparison of a porcine intestinal epithelial cell line and primary hepatocyte monocultures with a porcine enterohepatic co-culture system12

Abstract

This study was based on our previously developed double-layered enterohepatic co-culture system, composed of nontumorigenic porcine intestinal epithelial cell line (IPEC-J2) and primary culture of porcine hepatocytes. The anti-inflammatory effect of spent culture supernatant of Lactobacillus plantarum 2142 (Lp2142; 13.3%) and sodium n-butyrate (2 mM) was tested on IPEC-J2 and hepatocyte monocultures as well as on the gut-liver co-culture. To mimic inflammation, lipopolysaccharide (LPS; 1 and 10 μg/mL) was applied. Production of IL-8 and IL-6 was measured as a marker of inflammatory responses. The paracellular permeability of the intestinal epithelium was also monitored by fluoresceinisothiocyanate-labeled dextran 4 assay. Significant increase of IL-8 concentration was observed in the IPEC-J2 monoculture (P < 0.01) while the level of IL-6 was not changed following LPS treatment. Concentration of IL-8 and IL-6 was grown significantly in hepatocyte monocultures (P < 0.05 and P < 0.001) as well as in the co-culture after 10 μg/mL LPS treatment (P < 0.001 and P < 0.001). One microgram per milliliter LPS caused elevated IL-8 level in the co-culture (P < 0.001) and in the hepatocyte monoculture (P < 0.01), while it caused increased IL-6 level only in the hepatocytes (P < 0.001). Production of IL-8 was significantly decreased by butyrate in case of 1 μg/mL as well as 10 μg/mL LPS exposure in the co-culture (P < 0.001). Application of butyrate also reduced IL-6 level in the co-culture after 10 μg/mL LPS treatment (P < 0.01). Lactobacillus plantarum 2142 decreased IL-8 level after incubation with 1 μg/mL LPS (P < 0.001), while in case of 10 μg/mL LPS treatment only a marginal lowering in IL-8 (P = 0.064) release was measured. The IL-6 concentration was significantly reduced (P < 0.01 in case of 1 μg/mL LPS treatment) by Lp2142 in the co-culture. Contrarily, the elevated IL-8 and IL-6 level of hepatocytes has not been reduced in case of either butyrate or Lp2142 addition. The enterohepatic co-culture model offers a possibility for fast and reliable screening of new candidates against enteric inflammation, which are of special interest in porcine medicine and health management. According to our results, Lp2142 and butyrate both seem to be effective as anti-inflammatory agents in LPS-triggered inflammatory response, tested in the gut-liver co-culture model.

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

“…In our previous works, secreted bioactive metabolites of lactobacilli were proved to be effective in the treatment of intestinal inflammation on in vitro intestinal models [15,16]. We demonstrated that spent culture supernatant (13.3%) of Lp 2142 was effective in decreasing IL-8 and TNF-α mRNA level in IPEC-J2 cultures after LPS treatment.…”

“…For instance, chlorogenic acid is not reported to have significant bioavailability until it reaches the colon, where the microbiota rapidly hydrolyses it to liberate caffeic acid [9]. Metabolites of probiotic bacteria also have a proven anti-inflammatory effect [15,16]. Furthermore, unabsorbed polyphenols in the colon could influence the constitution and quality of microbiota.…”

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.…”

The Microbiome-Mitochondrion Connection: Common Ancestries, Common Mechanisms, Common Goals

“…The ability of specified microbiome lactobacilli to metabolize polyphenols found in certain fruits (16) or fermented soy (8), which work in concert with SCFAs such as butyrate (10, 20) to enhance muscle oxidative capacity by stimulating mitochondria, can potentially be exploited to produce a form of metabolic stabilization somewhat reminiscent of that exerted by physical activity. Finally, as advanced age is associated with both muscle loss (sarcopenia) and microbiome dybiosis, such a therapeutic approach holds the potential of slowing the onset of several metabolic and structural deficits inflicted by aging in elderly people.…”