Alteration in Inflammatory Responses and Cytochrome P450 Expression of Porcine Jejunal Cells by Drinking Water Supplements

Abstract

The intestinal epithelium is the first determining barrier to the drugs administered per os. Cytochrome P450 (CYP) enzymes are substantial in the initial step of xenobiotic metabolism; therefore, intestinal CYP enzyme activities could be an important influencing factor of the oral utilization of xenobiotic substances. In this study, the effect of four drinking water supplements on CYP mRNA levels of porcine intestinal epithelial cells was examined. Further goal of the study is to describe the effect of these feed additives on the proinflammatory response of the LPS-treated enterocytes. The nontransformed porcine intestinal epithelial cells (IPEC-J2) were grown on six-well polyester membrane inserts. Cell cultures were treated with LPS (10 μg/ml), β-glucan (5 and 50 μg/ml), sanguinarine-containing additive (5 and 50 μg/ml), drinking water acidifier (0.1 and 1 μl/ml), and fulvic acid (25 and 250 μg/ml) for 1 hour. Cells were washed with culture medium and incubated for additional 1 h before total RNA isolation. IL-6, IL-8, TNF-α, HSP70, CYP1A1, CYP1A2, and CYP3A29 mRNA levels were measured. The LPS treatment upregulated the gene expression of IL-8 and TNF-α. The relative gene expression of IL-6 remained unchanged and TNF-α and HSP70 were downregulated after the treatment with each feed additive. CYP1A1 and CYP1A2 expressions increased after sanguinarine-containing solution, fulvic acid, and drinking water acidifier treatment. None of the treatments changed the gene expression of CYP3A29, responsible for the metabolism of the majority of drug substances used in swine industry. The feed additive substances inhibited the expression of proinflammatory mediators HSP70 and TNF-α; however, β-glucan and fulvic acid elevated the production of the chemokine IL-8 mRNA in endotoxin-treated enterocytes. All acidic supplements increased the expression of CYP1A1 gene; their constituents may serve as a ligand of CYP1A1 nuclear receptors.

Sanguinarine protects against indomethacin-induced small intestine injury in rats by regulating the Nrf2/NF-κB pathways

“…Four principal isoquinoline alkaloids, such as SA, chelerythrine, allocryptopine, chelionine, were found in the MCE ( Huang et al, 2018 ). Among them, SA possesses strong anti-tumor, anti-inflammatory and antioxidant activities, as well as promoting animal growth ( Ahmad et al, 2000 ; Rahman et al, 2016 ; Palócz et al, 2019 ; Liu et al, 2020 ). SA is a potent inhibitor of NF-κB ( Chaturvedi et al, 1997 ), mitogen-activated protein kinase (MAPK) phosphatase-1 ( Vogt et al, 2005 ) and Akt signaling cascade ( Rahman et al, 2016 ), as well as a strong protective agent of HO-1 ( Park et al, 2014 ; Vrba et al, 2012 ).…”

“…SA is a potent inhibitor of NF-κB ( Chaturvedi et al, 1997 ), mitogen-activated protein kinase (MAPK) phosphatase-1 ( Vogt et al, 2005 ) and Akt signaling cascade ( Rahman et al, 2016 ), as well as a strong protective agent of HO-1 ( Park et al, 2014 ; Vrba et al, 2012 ). It has been reported that SA could inhibit acetic acid-induced ulcerative colitis by regulating the MAPK/NF-κB pathway in mice ( Niu et al, 2013 ), mitigate inflammatory responses in porcine jejunal cells ( Palócz et al, 2019 ), and enhance growth performance in broilers by modulating gut microbiome ( Liu et al, 2020 ). However, whether SA can relieve small intestinal inflammatory injury induced by NSAID has not been reported.…”

In vitro study of chlorine dioxide on porcine intestinal epithelial cell gene markers

“…Quantitative PCR was performed using the iQ SYBR Green Supermix kit (BioRad, Hercules, CA, USA) on the MiniOpticon System (BioRad) according to Palócz et al. ( 2019 ).…”

Preclinical safety evaluation of Macleaya Cordata extract: A re-assessment of general toxicity and genotoxicity properties in rodents

“…Previous studies tested the stability of SAN and CHE in an artificially simulated gastrointestinal environment and showed that SAN and CHE were stable in an acidic solution, but the average recoveries of SAN and CHE were only 60.53% and 87.89% in an alkaline environment at pH = 8 (Zhang et al, 2017). The mRNA expression of CYP1A1 and CYP1A2 was increased in porcine intestinal epithelial cells after the administration of 5 and 50 μg/ml SAN (Palócz et al, 2019). Current evidence suggests that CYP1A1 and CYP1A2 play an important role in the conversion of SAN to dihydrosanguinarine (DHSA, the first metabolite of SAN metabolic transformation) (Psotová et al, 2006;Deroussent et al, 2010;Sandor et al, 2018) and this is considered to be a possible detoxification process for SANs (Vrba et al, 2004).…”