The Integrins Involved in Soybean Agglutinin-Induced Cell Cycle Alterations in IPEC-J2

Abstract

Soybean agglutinin (SBA) is an anti-nutritional factor of soybean, affecting cell proliferation and inducing cytotoxicity. Integrins are transmembrane receptors, mediating a variety of cell biological processes. This research aims to study the effects of SBA on cell proliferation and cell cycle progression of the intestinal epithelial cell line from piglets (IPEC-J2), to identify the integrin subunits especially expressed in IPEC-J2s, and to analyze the functions of these integrins on IPEC-J2 cell cycle progression and SBA-induced IPEC-J2 cell cycle alteration. The results showed that SBA lowered cell proliferation rate as the cell cycle progression from G0/G1 to S phase (P < 0.05) was inhibited. Moreover, SBA lowered mRNA expression of cell cycle-related gene CDK4, Cyclin E and Cyclin D1 (P < 0.05). We successfully identified integrins α2, α3, α6, β1, and β4 in IPEC-J2s. These five subunits were crucial to maintain normal cell proliferation and cell cycle progression in IPEC-J2s. Restrain of either these five subunits by their inhibitors, lowered cell proliferation rate, and arrested the cells at G0/G1 phase of cell cycle (P < 0.05). Further analysis indicated that integrin α2, α6, and β1 were involved in the blocking of G0/G1 phase induced by SBA. In conclusion, these results suggested that SBA lowered the IPEC-J2 cell proliferation rate through the perturbation of cell cycle progression. Furthermore, integrins were important for IPEC-J2 cell cycle progression, and they were involved in the process of SBA-induced cell cycle progression alteration, which provide a basis for further revealing SBA anti-proliferation and anti-nutritional mechanism.

Eleutheroside B increase tight junction proteins and anti‐inflammatory cytokines expression in intestinal porcine jejunum epithelial cells (IPEC‐J2)

“…In addition, Schierack et al, (2006) reported an increased in the proliferation of IPEC-J2 cells after exposure to different Escherichia coli strains. Contrary to our findings, Pan et al (2013Pan et al ( , 2017 observed a decrease in the cell proliferation rate after treatment of IPEC-J2 cells with soybean agglutinin. In addition, Yoou, Kim, and Jeong (2015) reported that acteoside decreases the proliferation of thymic stromal lymphopoietin (TSLP)stimulated human mast cells.…”

“…The occurrence of antibiotic‐resistant strains among many pathogenic microorganisms and consumer requirement for organic food products has triggered the search for an alternative approach to control microbial infection in pigs (Adams et al, ; Che et al, ). The supplementation of various plant extracts in animal studies or in vitro studies has served as a promising alternative to antibiotic use (Che et al, ; Pan et al, ; Wang et al, ; Zhao et al, ). Understanding how these plant extracts modify the gastrointestinal milieus and nutritional requirements of pigs (Skjolaas, Burkey, Dritz, & Minton, ) and other animals is vital.…”

“…supplementation of various plant extracts in animal studies or in vitro studies has served as a promising alternative to antibiotic use(Che et al, 2018;Pan et al, 2017;Wang et al, 2016;Zhao et al, 2014).Understanding how these plant extracts modify the gastrointestinal milieus and nutritional requirements of pigs(Skjolaas, Burkey, Dritz, & Minton, 2007) and other animals is vital. Early weaning of piglets is a common husbandry practice in most commercial pig farms(Adams et al, 2018(Adams et al, , 2019, pigs during this condition has less developed gastrointestinal tracts(Chen et al, 2017) and reduce immunological protections, leading to the increase in disease susceptibility and infections by enteric bacteria(Adams et al, 2018;Kick, Tompkins, Flowers, Whisnant, & Almond, 2012).…”

“…In addition, the epithelial cell barrier protects the mucous membrane of the gastrointestinal tract(Pisal, Yellepeddi, Kumar, & Palakurthi, 2008).The tight intracellular junction proteins lie between the epithelial cells, which restrict the absorption of drugs and prevent the flow of possible antigens and pathogens from the lumen to the sub-mucosa(De Magistris, 2006;Pisal et al, 2008). However, the exposure of the intestinal epithelium to anti-nutritional factors and xenotoxins induced cytotoxicity to the intestinal epithelial cells(Pan et al, 2017), decreased intestinal cell viability(Pan et al, 2013), disrupts the brush border membrane(Zhao et al, 2011), and induced atrophy of the small intestinal microvilli of the epithelial cells(Pusztai et al, 1990).…”

N‐Acetyl‐d‐galactosamine prevents soya bean agglutinin‐induced intestinal barrier dysfunction in intestinal porcine epithelial cells

“…The release of alkaline phosphatase in the intestinal epithelial cells increases intestinal membrane permeability, elevating the occurrence of inflammation and infections (Bates, Akerlund, Mittge, & Guillemin, ; Geddes & Philpott, ), and decrease in TEER value. The increase in intestinal membrane permeability results in the reduction in tight junction integrity, disruption of cell‐to‐cell adhesions and decrease in cell viability and causes intestinal villus atrophy (Grant, Ewen et al, ; Pan et al, ; Pan, Zhao et al, ; Zhao et al, ), affecting cellular morphology, metabolism and proliferation (Pan et al, ). Several studies reported that the supplementation of SBA in intestinal epithelial cells decreased the TEER value, hence affecting membrane permeability (Dalla Pellegrina et al, ,; Pan et al, ).…”