The function of antimicrobial peptide MPX on the apoptosis and barrier of IPEC-J2 cells

The porcine intestinal epithelial cell line IPEC-J2 cells, which were isolated from neonatal piglet mid-jejunum, the main components of the intestinal epithelium and play an important role in the intestine. Escherichia coli is an important cause of diarrhea in human and animal. E.coli infections are the leading cause of travelers' diarrhea and a major cause of diarrhea in developing nations, where it can be life-threatening among children. The aim of this study is to explore MPX against E. coli to inhibit IPEC-J2 cells apoptosis and enhance cell barrier. In this study, scanning electron microscopy results found that E. coli infection caused cell apoptosis and destroyed cell membranes of IPEC-J2 cell. MPX effectively alleviated apoptosis of IPEC-J2 cells. The laser confocal results further found that MPX prevented cell apoptosis by inhibiting caspase-3 and caspase-9 activation. In addition, it was found that MPX regulated the expression of tight junction proteins ZO-1, Occludin, and Claudin-1 in IPEC-J2 cells and is closely related to Rac1 by adding a Rac1 inhibitor. The results of adhesion and invasion experiments further found that MPX inhibited bacterial adhesion and invasion through Rac1. The above results indicated that MPX has better function in inhibiting IPEC-J2 cells apoptosis and enhancing cell barrier.

Tryptophan improves porcine intestinal epithelial cell restitution through the CaSR/Rac1/PLC-γ1 signaling pathway

This study aimed to investigate the effect of tryptophan on cell migration and its underlying mechanism in porcine intestine epithelial cells (IPEC-J2). In this study, tryptophan can modulate IPEC-J2 cell...

Protective Effects of Fucoidan against Hydrogen Peroxide-Induced Oxidative Damage in Porcine Intestinal Epithelial Cells

This study was conducted to evaluate the effectiveness of fucoidan in ameliorating hydrogen peroxide (H2O2)-induced oxidative stress to porcine intestinal epithelial cell line (IPEC-1). The cell viability test was initially performed to screen out appropriate concentrations of H2O2 and fucoidan. After that, cells were exposed to H2O2 in the presence or absence of pre-incubation with fucoidan. Hydrogen peroxide increased the apoptotic and necrotic rate, boosted reactive oxygen species (ROS) generation, and disturbed the transcriptional expression of genes associated with antioxidant defense and apoptosis in IPEC-1 cells. Pre-incubation with fucoidan inhibited the increases in necrosis and ROS accumulation induced by H2O2. Consistently, in the H2O2-treated IPEC-1 cells, fucoidan normalized the content of reduced glutathione as well as the mRNA abundance of NAD(P)H quinone dehydrogenase 1 and superoxide dismutase 1 while it prevented the overproduction of malondialdehyde. Moreover, H2O2 stimulated the translocation of nuclear factor-erythroid 2-related factor-2 to the nucleus of IPEC-1 cells, but this increase was further promoted by fucoidan pre-treatment. The results suggest that fucoidan is effective in protecting IPEC-1 cells against oxidative damage induced by H2O2, which may help in developing appropriate strategies for maintaining the intestinal health of young piglets.

PSVI-37 Effects of deoxynivalenol (DON) and selected feed additives on viability and tight junction integrity of IPEC-J2 cells

Intestinal epithelial cells represent the frontline of host defense against invasion by enteric pathogens and uptake of potentially harmful substances. Mycotoxins are deleterious feed contaminants routinely found in cereal grains used in livestock feed. Development of strategies to eliminate mycotoxins and mitigate their harmful effects on the animal is of major importance to the livestock industry. The objective of the current study was to determine the effects of deoxynivalenol (DON), a common mycotoxin contaminant, and selected prebiotic, probiotic, and essential oil blended feed additives, on viability and integrity in porcine intestinal epithelial cell line (IPEC-J2). IPEC-J2 cells were grown on either plastic or semipermeable membranes until 100% confluence and treated with DON (12.5 uM) with or without additives (0.01 mg/mL Biolex® and 0.0001 mg/mL Encinnate®, Biomatrix Inc.) Use of DON at 12.5 uM decreased the trans-epithelial electrical resistance (TEER) by 37.5% compared to control, and led to 88.7% increase in FITC-dextran permeability after 3 days of culture, but with no negative effects on cell viability. The decrease in TEER by DON was minimized in the presence of Biolex (22.2%) and Encinnate (20.5%) relative to control. Treatment with DON also decreased the mRNA (24.0%) and protein abundance (64.4%) of claudin 3 and this was partially reversed by the additives, 11.5 and 11.0% decrease for claudin 3 mRNA and 58.7 and 56.7% decrease for the protein relative to control for Biolex and Encinnate, respectively. In summary, DON led to impaired epithelial tight junction integrity and this was partially rescued by the additives tested.

Protective Effect of Koumine, an Alkaloid from Gelsemium Sempervirens, on Injury Induced by H2O2 in IPEC-J2 Cells

Medicinal herbal plants have been commonly used for intervention in different diseases and improvement of health worldwide. Koumine, an alkaloid monomer found abundantly in Gelsemium plants, can be effectively used as an antioxidant. The purpose of this study was to evaluate the potential protective effect of koumine against hydrogen peroxide (H2O2)-induced oxidative stress and apoptosis in porcine intestinal epithelial cell line (IPEC-J2 cells). MTT assays showed that koumine significantly increased cell viability in H2O2-mediated IPEC-J2 cells. Preincubation with koumine ameliorated H2O2-medicated apoptosis by decreasing reactive oxygen species (ROS) production, and efficiently suppressed the lactate dehydrogenase (LDH) release and malondialdehyde (MDA) production. Moreover, a loss of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) activities was restored to normal level in H2O2-induced IPEC-J2 cells upon koumine exposure. Furthermore, pretreatment with koumine suppressed H2O2-mediated loss of mitochondrial membrane potential, caspase-9 and caspase-3 activation, decrease of Bcl-2 expression and elevation of Bax expressions. Collectively, the results of this study indicated that koumine possesses the cytoprotective effects in IPEC-J2 cells during exposure to H2O2 by suppressing production of ROS, inhibiting the caspase-3 activity and influencing the expression of Bax and Bcl-2. Koumine could potentially serve as a protective effect against H2O2-induced apoptosis.