Preliminary Investigation on the Effect of <i>Lactobacillus</i> and Epidermal Growth Factor on Tight Junction Proteins in Experimental Clostridium <i>difficile</i> Infection

Abstract

Clostridium difficile associated disease (CDAD) is the most common hospital acquired infection, due to exposure to various drugs. C. difficile toxins influence barrier function in intestinal epithelium. Biotherapeutic approaches, employing probiotic and epidermal growth factor (EGF) could help in barrier protein protection and aid in CDAD management. A preliminary investigation on the effect of Lactobacillus acidophilus and EGF on tight junction proteins in experimentally induced C. difficile infection was done. BALB/mice were divided into 5 groups. Group 1 was comprised of healthy controls, whereas animals in Groups 2 -5 were sub-divided into 3 subgroups (a, b and c) each. Animals in Groups 2 -5 received C. difficile inoculum either on day 1 (Group 2) or after pretreatment with ampicillin (Group 3), cyclosporine (Group 4) or lansoprazole (Group 5). Additionally animals in subgroups "b" and "c" also received L. acidophilus and EGF inocula respectively after C. difficile challenge. All animals were investigated for the presence of tight junction proteins (occludin, α-actinin and zonula occludens) in their colonic segments. Data were analyzed using the SPSS version 10 software. These three proteins were present in significantly less (P < 0.05) number of animals in the drug receiving animals, whereas they were found in significantly more (P < 0.05) number of animals receiving L. acidophilus and EGF after challenge with ampicillin, cyclosporine and lansoprazole, suggesting their role in protecting intestinal barrier function.

Effects of glucagon‐like peptides 1 and 2 and epidermal growth factor on the epithelial barrier of the rumen of adult sheep

“…In the published literature, the role of EGF is mainly seen in a protection of the EB against stressing conditions rather than directly improving the EB (Banan et al, 2003;Basuroy, Seth, Elias, Naren, & Rao, 2006;Guntaka, Samak, Seth, LaRusso, & Rao, 2011;Koepke et al, 2015;Okuyama et al, 2007;Sheth, Seth, Thangavel, Basuroy, & Rao, 2004). It often prevented or decreased the detrimental effect of micro-organisms on the epithelial barrier (Buret, Mitchell, Muench, & Scott, 2002;Kaur, Vaishnavi, Ray, Singh, & Kochhar, 2014;Lamb-Rosteski et al, 2008). EGF does so by stimulation of several intracellular pathways involved in TJ modulation.…”

Epidermal Growth Factor and Intestinal Barrier Function

“…The intestinal development is related to the intestinal barrier integrity directly, to keep intestinal health, and intracellular homeostasis is essential for the formation of the intestinal barrier. Pervious study has shown EGF controlling mucosal homeostasis through regulating the tight junction components [ 9 , 40 ], enhancing the mucins secretion [ 41 , 42 ], and decreasing pathogens colonization [ 8 , 27 – 30 ].…”

“…In addition, EGF can reduce C. jejuni colonization in the jejunum of C. jejuni infected chicks and prevent C. jejuni -induced claudin-4 disruption [ 30 ]. What is more, EGF showed a protective effect on TJs in experimental Clostridium difficile (C. difficile) infected mice [ 9 ], suggesting a potential role of EGF in reducing C. difficile colonization.…”

“…Dysfunction of the intestinal barrier is associated with increased gut permeability and development of multiple gastrointestinal diseases, such as food allergy, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), celiac disease, and infectious enterocolitis [ 2 – 4 ]. Recently, several substances, such as glutamine [ 2 ], sodium butyrate [ 5 ], bile acid [ 6 ], lactic acid bacteria [ 7 ], and epidermal growth factor (EGF) [ 8 , 9 ], have been reported to have a protective effect on intestinal epithelial through various underlying mechanisms.…”

“…In additional to enhancing cellular proliferation and differentiation, EGF also functions as a gastrointestinal tract (GI) mucosal protective factor, which associates with intestinal maturation and maintenance of epithelial cell homeostasis in the small intestine [ 17 ]. The biological actions of EGF are mediated via binding to the EGF receptor (EGFR), a transmembrane receptor tyrosine kinase of the ErbB family, that leads to autophosphorylation of receptor tyrosine kinase (RTK) and subsequent activation of Ras/mitogen-activated protein kinases (Ras/MAPK), phosphatidylinositol 3-kinase/AKT (PI3K/AKT), phospholipase C- γ /protein kinase C (PLC- γ /PKC), and STATS signal pathways [ 18 ], to promote intestinal development [ 15 , 19 – 22 ], regulate tight junction protein expression [ 9 , 23 – 25 ], reduce cell autophagy [ 26 ], inhibit apoptosis induced by oxidative stress [ 16 ], and reduce the colonization of the intestinal epithelium by enteropathogens [ 8 , 27 – 30 ].…”